Research ArticleSimultaneous Determination of Seven Active Components in RatPlasma by UHPLC-MSMS and Application to a QuantitativeStudy after Oral Administration of Huang-Lian Jie-Du Decoctionin High Fat-Induced Atherosclerosis Rats
1 Jiangxi Provincial Key Laboratory of TCM Etiopathogenesis Jiangxi University of Traditional Chinese MedicineNanchang 330004 China2Key Laboratory of Pharmacology of Traditional Chinese Medicine in Jiangxi Nanchang 330004 China3Pharmacy Department of Zhejiang Hospital Hangzhou 310013 China4Center for Health System and Policy Research Institute of Medical Information Chinese Academy of Medical SciencesBeijing 100020 China
Correspondence should be addressed to Guoliang Xu xuguoliang6606126com
Received 13 April 2019 Revised 23 May 2019 Accepted 10 June 2019 Published 1 July 2019
Academic Editor Samuel Carda-Broch
Copyright copy 2019 Li Jiang et al This is an open access article distributed under the Creative Commons Attribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited
Huang-Lian Jie-Du decoction (HLJDD) has been used to treat cardiovascular and cerebrovascular disease for many years in ChinaCurrently the determination of effect components inHLJDD is focusing either on the formula or on the extract while quantificationof that in biological samples is scarce especially simultaneous determination ofmulticomponent In this paper a rapid specific andsensitive ultra-high performance liquid chromatography-tandem mass spectrometry method was developed and fully validatedfor the simultaneous determination of seven main active constituents ie baicalin baicalein wogonoside wogonin berberinepalmatine jatrorrhizine in rat plasmaThemethod was also successfully applied to a quantitative study after oral administration ofHLJDD at different doses of 15 3 and 6 gkg body weight to high fat-induced atherosclerosis ratsThe analytes were detected by ESIsource andmultiple reactionsmonitoring (MRM) using positive scanningmodeThe bloodwas collected from the abdominal aortaof rats at predetermined time and preprepared with icariin and tetrahydropalmatine as internal standards (IS) Sample preparationwas achieved by protein precipitation (PPT)The validation parameters (linearity sensitivity intra-interday precision and accuracyextraction recovery and matrix effect) were within acceptable ranges and biological extracts were stable during the entire storingand preparing process And the result of determination ofHLJDD-containing plasma baicalin baicalein wogonoside andwogonincould be highly detected in a dose-dependent manner while berberine jatrorrhizine and palmatine were determined in a verylow level and in a dose-independent mode Thus the established method was sensitive enough and successfully applied to thedetermination of seven effective components in plasma taken from 24 high fat-induced atherosclerosis rats after oral administrationof three dosages of HLJDD
1 Introduction
Atherosclerosis (AS) is a chronic disease with a typicalpathological process of abnormal lipid metabolism ieinfiltrating into the arterial intima and depositing on thevessel wall which triggers inflammation and then in turnleads to the abnormal response to injury of the vessel
wall [1] such as alteration exudation and proliferationThese pathological changes are also prone to myocardialinfarction angina pectoris arrhythmia stroke and severesudden death A large number of clinical and basic studieshave shown that the risk factors for AS include hyperlipi-demia hypertension hyperglycemia obesity and metabolicdisorders of autologous bioactive substances [2] Although
HindawiInternational Journal of Analytical ChemistryVolume 2019 Article ID 5628160 12 pageshttpsdoiorg10115520195628160
2 International Journal of Analytical Chemistry
the pathogenesis of AS is not fully understood inflammation-injury-reaction theory proposed by Professor Ross [3] hasbeen widely recognized around the world The medicineused to treat AS currently is mostly anti-inflammatory drugsand it has become a new way to prevent and treat ASHowever there is still a lack of effective drugs for AS InTCM there are many theories according to the pathogen-esis of AS such as damp heat (Shi Re in Chinese) toxicpathogen (Du Xie in Chinese) and phlegm stasis (Tan Yuin Chinese) that is to say therapeutic principles of clearingaway heat and dampness and detoxicating would be effectivefor AS
Hang-Lian Jie-Du Decoction (HLJDD) a classic pre-scription of TCM was composed of four common herbsat the ratio of 3223 Rhizome Coptidis Radix ScutellariaeCortex Phellodendri and Fructus Gardeniae This formulafirst recorded by a doctor namedWang Tao in hismonographldquoWai Tai Mi Yaordquo (Chinese Tang Dynasty) had been usedfor 1700 years in China [4] and was mainly depending onclearing away heat-toxin to intervene in the inflammatorydisorder the basic pathophysiological process of AS [3]The existing research showed that HLJDD has a number ofpharmacological effects such as anti-inflammation loweringblood sugar regulating blood lipids and antioxidationThusit was used widely to treat diseases such as coronary heartdisease [5] diabetes [6] hypertension [7] angina [8] andmyocardial infarction [9] in clinic Recently accumulatingevidence has shown HLJDDrsquos efficacy on AS [10] and thecomponents in HLJDD have been extensively studied inorder to elucidate the mechanism For example berberinethe component of Rhizome Coptidis and Cortex Phellodendricould inhibit the synthesis of lipids by activating AMPenzymes and controlling LDL receptor expression [11] Theextracts of Radix Scutellariae could significantly reduce thecontent of oxidation of LDL and suppress inflammatoryresponses in macrophages [12] The effective ingredients inFructus Gardeniae could reduce the fat by inhibiting theactivity of pancreatic lipase [13] In addition alkaloids (egberberine palmatine and jatrorrhizine) flavonoids (egbaicalin baicalein wogonoside and wogonin) and iridoids(eg geniposide and shanzhiside) were themajor active com-ponents of HLJDD according to the previous phytochemicalresearch [14ndash16] Thus it was essential to quantify the aboveeffective components accurately in HLJDD treated for ASHowever the reported pharmacokinetic studies of HLJDDfailed to fully reflect the in vivo process of the compounddue to its limited components in recent years Moreover todate the multicomponents of this effective and traditionalformula in biological samples (eg AS rats) have been nearlyrarely provided so far Generally UHPLC-MSMS has beenregarded as one of the premier tools in complex biologicalsamples especially for properties of Chinese medicine for-mulae Therefore we developed a UHPLC-MSMS methodfor simultaneous determination of the above components inrat plasma Then the method was used to quantify the activeingredients (Figure 1) inAS rat plasma after three oral dosagesof HLJDD
2 Experimental
21Materials and Chemicals Themedicinalmaterials ofRhi-zome Coptidis Radix Scutellariae Cortex Phellodendri andFructus Gardeniae were purchased from Jiangxi JiangzhongTraditional ChineseMedicine Pieces Company and theywereauthenticated by Professor Shouwen Zhang (School of Phar-macy Jiangxi University of Traditional Chinese MedicineChina) Baicalin was obtained from National Institutes forFood and Drug Control Baicalein wogonoside wogoninberberine palmatine jatrorrhizine geniposide icariin andtetrahydropalmatine were purchased fromNational Pharma-ceutical Engineering Center for Solid Preparation in ChineseHerbal Medicine The purity of these standards was morethan 980 HPLC-grade methyl alcohol (MeOH) and ace-tonitrile (ACN)were obtained fromMerk (MerckGermany)and formic acid and ammonium formate were of chro-matographic purity and were purchased from DikmaPure(DikmaPure USA) and Sigma (Sigma USA) respectivelyDeionized water was prepared by a Millipore Alpha-Q waterpurification system (Millipore USA) Other chemicals andsolvents were all of analytical grade
22 Preparation of HLJJD An amount of crude drug equiv-alent to a daily dose of Huang-Lian-Jie-Du decoction wasweighed and the four medicinal herbs (Rhizome CoptidisRadix Scutellariae Cortex Phellodendri and Fructus Garde-niae)weremixed in the ratio 3 2 2 3Themedicinalmaterialswere soaked for 1 h and refluxed with water (16 wv) for30 min Filtrates were collected and the residues were thenrefluxed in water (14 wv) for 20min Two batches of filtrateswere combined and the obtained solution was concentratedby a rotatory vacuum evaporator concentrated to 05gmLcrude herb Then the extract was stored at 4∘C before use
23 Preparation of Stock and Working Solutions Accuratelyweigh a certain amount of baicalin baicalein wogonosidewogonin berberine palmatine jatrorrhizine icariin andtetrahydropalmatine in a 25mL volumetric flask dissolvedand diluted to the concentration of 144 92 176 140 200 116423 480 and 488 120583gmL respectively as the stock solutionsThen the stock solutions were further diluted into 210-26250200-17500 200-18750 200-1367 050-78125 050-56640050-250 800 and 50 ngmL respectively as the workingsolutions
24 Preparation of Calibration Standard and Quality Control(QC) Samples The calibration standard samples were pre-pared by freshly spiking the appropriate working solutioninto blank plasma yielding the concentrations of 21-2625 20-1750 20-1875 20-13672 025-7813 025-5664 and 025-25ngmL for baicalin baicalein wogonoside wogonin berber-ine palmatine and jatrorrhizine respectively and processedas described in the sample preparation Quality control(QC) samples used for the intra- and interday accuracyand precision extraction recovery and stability study wereprepared in the same way as calibration standard samples atconcentrations of 21 250 and 2625 ngml for baicalin 20
International Journal of Analytical Chemistry 3
OOH
HO
O O
O
OHHOOC
OH
OH
Baicalin
OOH
HO
HO O
Baicalein
OOH
O O
Wogonoside
O
OH
HO
HO
COOHOOH
HO O
Wogonin
O
O
Berberine Palmatine
HO
Jatrorrhizine
O
O
O
O
OH
OH
OH
OH
OO
OH
OH
HO
Icariin
N
Tetrahydropalmatine
OCH3 OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
N+
H3CO
H3CO
H3CO
N+
H3C
HOH2C
N+
H3CO
H3CO
Figure 1 Chemical structures of baicalin baicalein wogonoside wogonin berberine palmatine jatrorrhizine icariin (IS) andtetrahydropalmatine (IS)
4 International Journal of Analytical Chemistry
250 and 575 ngml for baicalein 20 250 and 1875 ngmlfor wogonoside 20 257 and 13672 ngml for wogoninand 025 25 and 250 ngml for berberine palmatine andjatrorrhizine
25 Plasma Sample Preparation An aliquot of 100120583L thawedplasma sample was transferred into an Eppendorf tube (EPtube) to which 10 120583L of seven analytes and internal standardssolution from each working solution were added After beingvortexed for 15 s 100 120583L methanol plus 300 120583L acetonitrilewas added and rested for 3 h following vortex mixing for30 s to precipitate protein Subsequently the mixture wascentrifuged at 15000timesg for 10 min 400 120583L supernatant wastransferred into another EP tube and evaporated to drynessunder the stream of nitrogen in a water bath at 40∘C Theresidue was dissolved in 100 120583L of reconstituted solutionwhich consisted of acetonitrile-01 formic acid (91 vv) andthen centrifuged at 18000timesg for 15 min after vortexing for 2min The supernatant was injected into the UHPLCndashMSMSsystem for analysis
26 Instruments and Chromatographic Conditions The anal-ysis was performed using the Shimadzu UHPLC system(Shimadzu Corporation Kyoto Japan) consisting of an LC-30AD binary pump a DGU-20A5 degassing unit a SIL-30AC autosampler and a CTO-30A5R column oven Massspectrometric detectionwas conducted on anAB SciexQtrap5500 System (Applied Biosystems Foster City CA USA)and equipped with Analyst software (version 162) for dataprocessing Chromatographic separation was achieved ona Shimadzu Shim-pack XR-ODS III (16 120583m 20 mmtimes75mm ShimadzuCorporation Kyoto Japan)Themobile phaseconsisted of 5 mM ammonium formate-01 formic acid(solvent A) and acetonitrile (solvent B) The gradient elutionconditions were optimized as follows 10-20 B (0-3 min)20-45 B (3-45 min) 45 B (45-7 min) 45-80 B(7-12min) 80-95 B (12-14 min) 95-10B (15-155 min) and 10B (155-16 min) The flow rate was set at 03mLmin withthe column temperature maintained at 40∘C The injectionvolume of 1 120583L was used for the reference standards andsamples
For mass detection the electrospray ionization sourcewas operated in positive mode The operating parameterswere optimized under the following conditions 500∘C forthe interface temperature and 55 kV for the ion sprayvoltage ion source gas 1 and gas 2 were fixed at 50 psiVacuum was obtained by a Turbo molecular pump (AgilentTechnologies USA) Nitrogen generated by the high puritynitrogen generator (99999 Peak Scientific InstrumentsLtd UK) was used as the source of curtain gas (30 psi)and collision gas (7 psi) The optimized multiple reactionmonitoring (MRM) parameters including collision energyand declustering potential are listed in Table 1
27 Validation of the UPLCndashMSMS Method The proposedmethod was validated for specificity extraction recovery
matrix effect LLOQ linearity and stabilityMeanwhile intra-and interday validation were performed to evaluate theaccuracy and precision of the measurements
271 Specificity Linearity Lower Limits of Detection andQuantification To investigate the specificity of this methodchromatogram comparison of blank plasma blank plasmaspiked with ISanalyte and rat plasma samples were con-ducted to assay for the exclusion of any endogenous interfer-ence existing at or close to the expected retention time of theanalytes Calibration curves were established from peak arearatios (analyte peak area to the internal standard peak areaAsAi) versus nominal concentrations using a linear least-squares regression model (1X2 weighting)
272 Precision and Accuracy Intra- and interday precisionsand accuracy were denoted by assessing measured results ofQC samples at low medium and high concentrations Eachof the five samples was processed in parallel Continuouslymeasure a batch of samples and calculate intraday precisionThen five samples of low medium and high plasma samplesof each componentwere prepared in parallel on different con-secutive days and the 45 samples were tested in three batchesand the interday precision was calculated by referring to theaccompanying standard Precisions were expressed by therelative standard deviation (RSD ) while accuracy () waspresented as the percentage difference between the meanmeasured concentrations and the spiked concentrations
273 Extraction Recovery and Matrix Effect Extractionrecoveries of the seven analytes from rat blank plasmawere determined by comparing the mean peak areas of theQC samples spiked before protein precipitation with thosespiked after protein extraction Matrix effects occurred whenendogenous molecules were coeluting with the analytes ofinterest enhancing or decreasing the ionization efficiency ofthe electrospray interface The matrix effect was assessed viacomparing the mean peak areas of the QC samples spikedafter the pretreatment with those of the pure solution
274 Stability Stability of seven active constituents waschecked by comparing measured results with those of freshlyprepared samples The short- and long-term stabilities wereevaluated by analyzing QC plasma samples kept at roomtemperature for 4 h and in the freezer (minus20∘C) for 30 daysrespectively the freeze-thaw stability was carried out bydetecting QC samples undergoing three freeze-thaw cyclesthe postpreparation stability was assessed by determining theextracted QC samples stored under autosampler conditions(4∘C) for 12 h
28 Determination of HLJDD-Containing Plasma Specifi-cally pathogen-free Sprague-Dawley rats (male weighing200 plusmn 20 g) were purchased from Hunan Slac LaboratoryAnimal Co LTD (Hunan China Certificate No SCXK-2013-0004) and acclimated in Exhaust Ventilated Closed-SystemCage Rack (EVC) for at least a week with environmentallycontrolled quarters (22plusmn2∘C and 1212-h lightdark cycle) and
International Journal of Analytical Chemistry 5
Table 1 Optimized precursorproduction pairs and multiple reaction monitoring (MRM) parameters for the analytes and IS
free access to standard chow and water Animal welfare andexperimental procedures were strictly in accordance withthe guide for the care and use of laboratory animal by theAnimal Ethics Committee of Jiangxi University of TCMAfter one week of acclimatization the mice were randomlydivided into five groups (119899=12) normal control group modelgroup and three dosage groups The normal control groupwas fed with common diet while the other groups werefed with high-fat diet (3 cholesterol 05 sodium cholate02 propylthiouracil 5 sugar 10 lard and 813 basicdiet) The high fat-induced AS rat model was established inour previous study [17 18] Briefly the AS model was madeby the combination of regular intraperitoneal injection ofvitamin D
3and high fat diet for 8 weeks The model rat
was injected with 600000 UKg vitamin D3on the second
week and 200000 UKg every other week The three dosagegroups were gavaged HLJDD at doses of 15 3 and 6 g kg(low medium and high) once daily from the 3rd week for8 weeks until they were sacrificed Then the heparinizedblood samples were collected from abdominal aorta after 30minutes of the last administration and frozen at minus20∘C untilanalysis
3 Results
31 Method Validation
311 Specificity The total separation time for all analyteswas 16 minutes and there were little interferential substanceswith the analytes and IS in the blank plasma Representativechromatogram of analytes and IS in rat plasma was shown inFigure 2
312 Linearity The calibration curve of each analyte wasestablished with at least six points of standard solution andeach point was repeated five times The calibration curvesof all analytes exhibited good linearity and the regressionequations with correlation coefficients and linear range werelisted in Table 2
313 Precision and Accuracy The intraday and interdayprecision of all analytes were all less than 15 whilst theaccuracy deviation values were all within 964plusmn60 of theactual values at each QC level (shown in Table 3)
314 Extraction Recovery and Matrix Effect The extractionrecoveries (absolute recoveries) of each component weremore than 80 at each QC level which satisfied the quan-titative requirements of biological samples With respect tomatrix effect no suppressive or enhancing effect was foundon the analytes and IS That is to say the responses of allcomponents in the matrix were consistent with that in puresolution The results are shown in Table 4
315 Stability Results of the stability (shown in Table 5)illustrate that all analytes remained generally stable in plasmafor 4 h when stored at room temperature or 30 days whenstored atminus20∘C for three freeze-thaw cycles And they showedsatisfactory stability in the reconstituted solutions whenstored under autosampler condition for 12 h
316 Application The present method was successfully usedfor the determination of three dosages of HLJDD in 24 highfat-induced AS rat plasmasThe concentration of the analytesin plasma after ig administration was shown in Table 6 andFigure 3
4 Discussion
41 Optimization of LCndashMS for Quantitative Analysis Thechoice of mobile phase was a crucial factor in achievingfine chromatographic behavior and appropriate ionizationModifiers such as formic acid and ammonium formatealone or in combination with different concentrations werecomparedThe best peak shape and ionization were achievedadapting 5 mM ammonium formate buffer Linear gradientelution was used to elute endogenous substances residuefrom the column In addition all analytes and IS were bothfully scanned by positive and negative mode As alkaloidswere detected overwhelmingly in the positive mode whileflavonoids with little difference in both modes the positivemode was used in the MRM acquisition
42 Selection of the Determined Components According tothe previous phytochemical and HPLCndashMS studies iridoidsalkaloids and flavonoids were the predominant constituentsin HLJDD [19ndash23] And many components of HLJDD could
6 International Journal of Analytical Chemistry
2 4 6 80
110
2 4 6 80
250
2 4 6 80
180
2 4 6 80
150
palmatine
jateorhizine
icariin
tetrahydropalmatine
2 4 6 80
85
2 4 6 80
225
2 4 6 80
275
2 4 6 80
300
2 4 6 80
350
baicalin
baicalein
wogonoside
wogonin
berberine
Inte
nsity
(CPS
)
Time (min)
(a)
2 4 6 80
12000
2 4 6 80
70000
2 4 6 80
2750
2 4 6 80
18000
54
58
54
56
2 4 6 80
14000
2 4 6 80
4500
2 4 6 80
40000
2 4 6 80
50000
2 4 6 80
650000
58
59
58
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(b)
2 4 6 80
4250
2 4 6 80
1100
2 4 6 80
850000
2 4 6 80
1700000
56
54
58
54
2 4 6 80
110000
2 4 6 80
10000
2 4 6 80
750000
2 4 6 80
22500
2 4 6 80
6500
58
58
59
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(c)
Figure 2 The MRM spectrum of each component (a) blank plasma (b) blank plasma spiked with analytes and IS (c) rat plasma samplecollected after ig administration of HLJDD
alleviate AS development by inhibiting the vascular inflam-matory processes which was the initiation and progressionof AS [3 4] The flavonoids of Radix Scutellariae iebaicalin baicalein and wogonin could suppress vascularinflammation in vitro and in vivo [24] andwogonoside couldmodulate inflammatory mediator expression in LPS-inducedRAW2647 cells [25] In addition wogonin inhibited MMP-9 gene expression (a major role in the pathogenesis of AS)
via MAPK signaling pathways [26] Berberine palmatineand jatrorrhizine the main alkaloids constituents in Rhi-zome Coptidis and Cortex Phellodendri could suppress theformation and development of AS by altering gut microbiotacompositions [27] anti-inflammation and lowering bloodlipids [28ndash30] Thus baicalin baicalein wogonoside wogo-nin berberine palmatine and jatrorrhizine were determinedin high fat-induced AS rats
International Journal of Analytical Chemistry 7
Baicali
n
Baicale
in
Wogonosid
e
Wogonin
Berberi
ne
Palmati
ne
Jatrorrh
izine
01
220
4020
0040
00C
once
ntra
tion
(ng
mL)
Low
Medium
High
Figure 3 The concentration of seven analytes in rat plasma after oral administration of three dosages of HLJDD
Table 2 Regression data and LLOQs of the multi-components determined in HLJDD
Although the determination of HLJDD was reportedbefore [31 32] the active components of it in biologicalsamples have seldom been reported Deng and He [33 34]determined baicalin and wogonoside in type 2 diabetic andnormal rats Zeng and Zhu [35 36] quantified baicalingeniposide and berberine in MCAO rats Nevertheless theywere all based on HPLCmethod and the LLOQwere 120583g levelfor those constituents Thus we quantified the above seveningredients simultaneously in the plasma of high fat-inducedAS rats after oral administration ofHLJDD except geniposide(for there was almost no geniposide in the drug-containingplasma and the pretreatment of geniposide was extremelyunstable) by UHPLC-MSMS In our study the precisionaccuracy matrix effect and stability under all conditionsare within bioanalytical methodology validation acceptancecriteria [37] with the extraction recovery of palmatine andberberine higher than Lursquos study [15] the LLOQ was lower
and the retention time greatly shortened than the previousresearch [33ndash36] Although some peaks eluted with closedretention time the MS detector can determine them accu-rately by taking the advantage of its high selective MRMmethod
43 Concentration Profiles of theAnalytes inASPlasma Fromthe result of determination of HLJDD-containing plasmabaicalin baicalein wogonoside andwogonin could be highlydetected in a dose-dependent manner while berberine jatr-orrhizine and palmatine were determined in a very lowlevel and in a dose-independent mode For one thing theformer absorptions were relatively better than the latterFor another flavonoids are easily bound to glucuronic acidor sulfuric acid to form two-phase metabolism so theirplasma concentration-time curves showed obvious bimodalphenomena and concentration increased slowly from the
8 International Journal of Analytical Chemistry
Table 3 Intra-inter-day precision and accuracy for the determination of the components in rat plasma
Analytes Spiked Concentration(ngmL)
Precision () Accuracy ( Mean plusmn SD)Intra-day Inter-day Intra-day Inter-day
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
SpectroscopyAnalytical ChemistryInternational Journal of
Hindawiwwwhindawicom Volume 2018
MaterialsJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
BioMed Research International Electrochemistry
International Journal of
Hindawiwwwhindawicom Volume 2018
Na
nom
ate
ria
ls
Hindawiwwwhindawicom Volume 2018
Journal ofNanomaterials
Submit your manuscripts atwwwhindawicom
2 International Journal of Analytical Chemistry
the pathogenesis of AS is not fully understood inflammation-injury-reaction theory proposed by Professor Ross [3] hasbeen widely recognized around the world The medicineused to treat AS currently is mostly anti-inflammatory drugsand it has become a new way to prevent and treat ASHowever there is still a lack of effective drugs for AS InTCM there are many theories according to the pathogen-esis of AS such as damp heat (Shi Re in Chinese) toxicpathogen (Du Xie in Chinese) and phlegm stasis (Tan Yuin Chinese) that is to say therapeutic principles of clearingaway heat and dampness and detoxicating would be effectivefor AS
Hang-Lian Jie-Du Decoction (HLJDD) a classic pre-scription of TCM was composed of four common herbsat the ratio of 3223 Rhizome Coptidis Radix ScutellariaeCortex Phellodendri and Fructus Gardeniae This formulafirst recorded by a doctor namedWang Tao in hismonographldquoWai Tai Mi Yaordquo (Chinese Tang Dynasty) had been usedfor 1700 years in China [4] and was mainly depending onclearing away heat-toxin to intervene in the inflammatorydisorder the basic pathophysiological process of AS [3]The existing research showed that HLJDD has a number ofpharmacological effects such as anti-inflammation loweringblood sugar regulating blood lipids and antioxidationThusit was used widely to treat diseases such as coronary heartdisease [5] diabetes [6] hypertension [7] angina [8] andmyocardial infarction [9] in clinic Recently accumulatingevidence has shown HLJDDrsquos efficacy on AS [10] and thecomponents in HLJDD have been extensively studied inorder to elucidate the mechanism For example berberinethe component of Rhizome Coptidis and Cortex Phellodendricould inhibit the synthesis of lipids by activating AMPenzymes and controlling LDL receptor expression [11] Theextracts of Radix Scutellariae could significantly reduce thecontent of oxidation of LDL and suppress inflammatoryresponses in macrophages [12] The effective ingredients inFructus Gardeniae could reduce the fat by inhibiting theactivity of pancreatic lipase [13] In addition alkaloids (egberberine palmatine and jatrorrhizine) flavonoids (egbaicalin baicalein wogonoside and wogonin) and iridoids(eg geniposide and shanzhiside) were themajor active com-ponents of HLJDD according to the previous phytochemicalresearch [14ndash16] Thus it was essential to quantify the aboveeffective components accurately in HLJDD treated for ASHowever the reported pharmacokinetic studies of HLJDDfailed to fully reflect the in vivo process of the compounddue to its limited components in recent years Moreover todate the multicomponents of this effective and traditionalformula in biological samples (eg AS rats) have been nearlyrarely provided so far Generally UHPLC-MSMS has beenregarded as one of the premier tools in complex biologicalsamples especially for properties of Chinese medicine for-mulae Therefore we developed a UHPLC-MSMS methodfor simultaneous determination of the above components inrat plasma Then the method was used to quantify the activeingredients (Figure 1) inAS rat plasma after three oral dosagesof HLJDD
2 Experimental
21Materials and Chemicals Themedicinalmaterials ofRhi-zome Coptidis Radix Scutellariae Cortex Phellodendri andFructus Gardeniae were purchased from Jiangxi JiangzhongTraditional ChineseMedicine Pieces Company and theywereauthenticated by Professor Shouwen Zhang (School of Phar-macy Jiangxi University of Traditional Chinese MedicineChina) Baicalin was obtained from National Institutes forFood and Drug Control Baicalein wogonoside wogoninberberine palmatine jatrorrhizine geniposide icariin andtetrahydropalmatine were purchased fromNational Pharma-ceutical Engineering Center for Solid Preparation in ChineseHerbal Medicine The purity of these standards was morethan 980 HPLC-grade methyl alcohol (MeOH) and ace-tonitrile (ACN)were obtained fromMerk (MerckGermany)and formic acid and ammonium formate were of chro-matographic purity and were purchased from DikmaPure(DikmaPure USA) and Sigma (Sigma USA) respectivelyDeionized water was prepared by a Millipore Alpha-Q waterpurification system (Millipore USA) Other chemicals andsolvents were all of analytical grade
22 Preparation of HLJJD An amount of crude drug equiv-alent to a daily dose of Huang-Lian-Jie-Du decoction wasweighed and the four medicinal herbs (Rhizome CoptidisRadix Scutellariae Cortex Phellodendri and Fructus Garde-niae)weremixed in the ratio 3 2 2 3Themedicinalmaterialswere soaked for 1 h and refluxed with water (16 wv) for30 min Filtrates were collected and the residues were thenrefluxed in water (14 wv) for 20min Two batches of filtrateswere combined and the obtained solution was concentratedby a rotatory vacuum evaporator concentrated to 05gmLcrude herb Then the extract was stored at 4∘C before use
23 Preparation of Stock and Working Solutions Accuratelyweigh a certain amount of baicalin baicalein wogonosidewogonin berberine palmatine jatrorrhizine icariin andtetrahydropalmatine in a 25mL volumetric flask dissolvedand diluted to the concentration of 144 92 176 140 200 116423 480 and 488 120583gmL respectively as the stock solutionsThen the stock solutions were further diluted into 210-26250200-17500 200-18750 200-1367 050-78125 050-56640050-250 800 and 50 ngmL respectively as the workingsolutions
24 Preparation of Calibration Standard and Quality Control(QC) Samples The calibration standard samples were pre-pared by freshly spiking the appropriate working solutioninto blank plasma yielding the concentrations of 21-2625 20-1750 20-1875 20-13672 025-7813 025-5664 and 025-25ngmL for baicalin baicalein wogonoside wogonin berber-ine palmatine and jatrorrhizine respectively and processedas described in the sample preparation Quality control(QC) samples used for the intra- and interday accuracyand precision extraction recovery and stability study wereprepared in the same way as calibration standard samples atconcentrations of 21 250 and 2625 ngml for baicalin 20
International Journal of Analytical Chemistry 3
OOH
HO
O O
O
OHHOOC
OH
OH
Baicalin
OOH
HO
HO O
Baicalein
OOH
O O
Wogonoside
O
OH
HO
HO
COOHOOH
HO O
Wogonin
O
O
Berberine Palmatine
HO
Jatrorrhizine
O
O
O
O
OH
OH
OH
OH
OO
OH
OH
HO
Icariin
N
Tetrahydropalmatine
OCH3 OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
N+
H3CO
H3CO
H3CO
N+
H3C
HOH2C
N+
H3CO
H3CO
Figure 1 Chemical structures of baicalin baicalein wogonoside wogonin berberine palmatine jatrorrhizine icariin (IS) andtetrahydropalmatine (IS)
4 International Journal of Analytical Chemistry
250 and 575 ngml for baicalein 20 250 and 1875 ngmlfor wogonoside 20 257 and 13672 ngml for wogoninand 025 25 and 250 ngml for berberine palmatine andjatrorrhizine
25 Plasma Sample Preparation An aliquot of 100120583L thawedplasma sample was transferred into an Eppendorf tube (EPtube) to which 10 120583L of seven analytes and internal standardssolution from each working solution were added After beingvortexed for 15 s 100 120583L methanol plus 300 120583L acetonitrilewas added and rested for 3 h following vortex mixing for30 s to precipitate protein Subsequently the mixture wascentrifuged at 15000timesg for 10 min 400 120583L supernatant wastransferred into another EP tube and evaporated to drynessunder the stream of nitrogen in a water bath at 40∘C Theresidue was dissolved in 100 120583L of reconstituted solutionwhich consisted of acetonitrile-01 formic acid (91 vv) andthen centrifuged at 18000timesg for 15 min after vortexing for 2min The supernatant was injected into the UHPLCndashMSMSsystem for analysis
26 Instruments and Chromatographic Conditions The anal-ysis was performed using the Shimadzu UHPLC system(Shimadzu Corporation Kyoto Japan) consisting of an LC-30AD binary pump a DGU-20A5 degassing unit a SIL-30AC autosampler and a CTO-30A5R column oven Massspectrometric detectionwas conducted on anAB SciexQtrap5500 System (Applied Biosystems Foster City CA USA)and equipped with Analyst software (version 162) for dataprocessing Chromatographic separation was achieved ona Shimadzu Shim-pack XR-ODS III (16 120583m 20 mmtimes75mm ShimadzuCorporation Kyoto Japan)Themobile phaseconsisted of 5 mM ammonium formate-01 formic acid(solvent A) and acetonitrile (solvent B) The gradient elutionconditions were optimized as follows 10-20 B (0-3 min)20-45 B (3-45 min) 45 B (45-7 min) 45-80 B(7-12min) 80-95 B (12-14 min) 95-10B (15-155 min) and 10B (155-16 min) The flow rate was set at 03mLmin withthe column temperature maintained at 40∘C The injectionvolume of 1 120583L was used for the reference standards andsamples
For mass detection the electrospray ionization sourcewas operated in positive mode The operating parameterswere optimized under the following conditions 500∘C forthe interface temperature and 55 kV for the ion sprayvoltage ion source gas 1 and gas 2 were fixed at 50 psiVacuum was obtained by a Turbo molecular pump (AgilentTechnologies USA) Nitrogen generated by the high puritynitrogen generator (99999 Peak Scientific InstrumentsLtd UK) was used as the source of curtain gas (30 psi)and collision gas (7 psi) The optimized multiple reactionmonitoring (MRM) parameters including collision energyand declustering potential are listed in Table 1
27 Validation of the UPLCndashMSMS Method The proposedmethod was validated for specificity extraction recovery
matrix effect LLOQ linearity and stabilityMeanwhile intra-and interday validation were performed to evaluate theaccuracy and precision of the measurements
271 Specificity Linearity Lower Limits of Detection andQuantification To investigate the specificity of this methodchromatogram comparison of blank plasma blank plasmaspiked with ISanalyte and rat plasma samples were con-ducted to assay for the exclusion of any endogenous interfer-ence existing at or close to the expected retention time of theanalytes Calibration curves were established from peak arearatios (analyte peak area to the internal standard peak areaAsAi) versus nominal concentrations using a linear least-squares regression model (1X2 weighting)
272 Precision and Accuracy Intra- and interday precisionsand accuracy were denoted by assessing measured results ofQC samples at low medium and high concentrations Eachof the five samples was processed in parallel Continuouslymeasure a batch of samples and calculate intraday precisionThen five samples of low medium and high plasma samplesof each componentwere prepared in parallel on different con-secutive days and the 45 samples were tested in three batchesand the interday precision was calculated by referring to theaccompanying standard Precisions were expressed by therelative standard deviation (RSD ) while accuracy () waspresented as the percentage difference between the meanmeasured concentrations and the spiked concentrations
273 Extraction Recovery and Matrix Effect Extractionrecoveries of the seven analytes from rat blank plasmawere determined by comparing the mean peak areas of theQC samples spiked before protein precipitation with thosespiked after protein extraction Matrix effects occurred whenendogenous molecules were coeluting with the analytes ofinterest enhancing or decreasing the ionization efficiency ofthe electrospray interface The matrix effect was assessed viacomparing the mean peak areas of the QC samples spikedafter the pretreatment with those of the pure solution
274 Stability Stability of seven active constituents waschecked by comparing measured results with those of freshlyprepared samples The short- and long-term stabilities wereevaluated by analyzing QC plasma samples kept at roomtemperature for 4 h and in the freezer (minus20∘C) for 30 daysrespectively the freeze-thaw stability was carried out bydetecting QC samples undergoing three freeze-thaw cyclesthe postpreparation stability was assessed by determining theextracted QC samples stored under autosampler conditions(4∘C) for 12 h
28 Determination of HLJDD-Containing Plasma Specifi-cally pathogen-free Sprague-Dawley rats (male weighing200 plusmn 20 g) were purchased from Hunan Slac LaboratoryAnimal Co LTD (Hunan China Certificate No SCXK-2013-0004) and acclimated in Exhaust Ventilated Closed-SystemCage Rack (EVC) for at least a week with environmentallycontrolled quarters (22plusmn2∘C and 1212-h lightdark cycle) and
International Journal of Analytical Chemistry 5
Table 1 Optimized precursorproduction pairs and multiple reaction monitoring (MRM) parameters for the analytes and IS
free access to standard chow and water Animal welfare andexperimental procedures were strictly in accordance withthe guide for the care and use of laboratory animal by theAnimal Ethics Committee of Jiangxi University of TCMAfter one week of acclimatization the mice were randomlydivided into five groups (119899=12) normal control group modelgroup and three dosage groups The normal control groupwas fed with common diet while the other groups werefed with high-fat diet (3 cholesterol 05 sodium cholate02 propylthiouracil 5 sugar 10 lard and 813 basicdiet) The high fat-induced AS rat model was established inour previous study [17 18] Briefly the AS model was madeby the combination of regular intraperitoneal injection ofvitamin D
3and high fat diet for 8 weeks The model rat
was injected with 600000 UKg vitamin D3on the second
week and 200000 UKg every other week The three dosagegroups were gavaged HLJDD at doses of 15 3 and 6 g kg(low medium and high) once daily from the 3rd week for8 weeks until they were sacrificed Then the heparinizedblood samples were collected from abdominal aorta after 30minutes of the last administration and frozen at minus20∘C untilanalysis
3 Results
31 Method Validation
311 Specificity The total separation time for all analyteswas 16 minutes and there were little interferential substanceswith the analytes and IS in the blank plasma Representativechromatogram of analytes and IS in rat plasma was shown inFigure 2
312 Linearity The calibration curve of each analyte wasestablished with at least six points of standard solution andeach point was repeated five times The calibration curvesof all analytes exhibited good linearity and the regressionequations with correlation coefficients and linear range werelisted in Table 2
313 Precision and Accuracy The intraday and interdayprecision of all analytes were all less than 15 whilst theaccuracy deviation values were all within 964plusmn60 of theactual values at each QC level (shown in Table 3)
314 Extraction Recovery and Matrix Effect The extractionrecoveries (absolute recoveries) of each component weremore than 80 at each QC level which satisfied the quan-titative requirements of biological samples With respect tomatrix effect no suppressive or enhancing effect was foundon the analytes and IS That is to say the responses of allcomponents in the matrix were consistent with that in puresolution The results are shown in Table 4
315 Stability Results of the stability (shown in Table 5)illustrate that all analytes remained generally stable in plasmafor 4 h when stored at room temperature or 30 days whenstored atminus20∘C for three freeze-thaw cycles And they showedsatisfactory stability in the reconstituted solutions whenstored under autosampler condition for 12 h
316 Application The present method was successfully usedfor the determination of three dosages of HLJDD in 24 highfat-induced AS rat plasmasThe concentration of the analytesin plasma after ig administration was shown in Table 6 andFigure 3
4 Discussion
41 Optimization of LCndashMS for Quantitative Analysis Thechoice of mobile phase was a crucial factor in achievingfine chromatographic behavior and appropriate ionizationModifiers such as formic acid and ammonium formatealone or in combination with different concentrations werecomparedThe best peak shape and ionization were achievedadapting 5 mM ammonium formate buffer Linear gradientelution was used to elute endogenous substances residuefrom the column In addition all analytes and IS were bothfully scanned by positive and negative mode As alkaloidswere detected overwhelmingly in the positive mode whileflavonoids with little difference in both modes the positivemode was used in the MRM acquisition
42 Selection of the Determined Components According tothe previous phytochemical and HPLCndashMS studies iridoidsalkaloids and flavonoids were the predominant constituentsin HLJDD [19ndash23] And many components of HLJDD could
6 International Journal of Analytical Chemistry
2 4 6 80
110
2 4 6 80
250
2 4 6 80
180
2 4 6 80
150
palmatine
jateorhizine
icariin
tetrahydropalmatine
2 4 6 80
85
2 4 6 80
225
2 4 6 80
275
2 4 6 80
300
2 4 6 80
350
baicalin
baicalein
wogonoside
wogonin
berberine
Inte
nsity
(CPS
)
Time (min)
(a)
2 4 6 80
12000
2 4 6 80
70000
2 4 6 80
2750
2 4 6 80
18000
54
58
54
56
2 4 6 80
14000
2 4 6 80
4500
2 4 6 80
40000
2 4 6 80
50000
2 4 6 80
650000
58
59
58
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(b)
2 4 6 80
4250
2 4 6 80
1100
2 4 6 80
850000
2 4 6 80
1700000
56
54
58
54
2 4 6 80
110000
2 4 6 80
10000
2 4 6 80
750000
2 4 6 80
22500
2 4 6 80
6500
58
58
59
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(c)
Figure 2 The MRM spectrum of each component (a) blank plasma (b) blank plasma spiked with analytes and IS (c) rat plasma samplecollected after ig administration of HLJDD
alleviate AS development by inhibiting the vascular inflam-matory processes which was the initiation and progressionof AS [3 4] The flavonoids of Radix Scutellariae iebaicalin baicalein and wogonin could suppress vascularinflammation in vitro and in vivo [24] andwogonoside couldmodulate inflammatory mediator expression in LPS-inducedRAW2647 cells [25] In addition wogonin inhibited MMP-9 gene expression (a major role in the pathogenesis of AS)
via MAPK signaling pathways [26] Berberine palmatineand jatrorrhizine the main alkaloids constituents in Rhi-zome Coptidis and Cortex Phellodendri could suppress theformation and development of AS by altering gut microbiotacompositions [27] anti-inflammation and lowering bloodlipids [28ndash30] Thus baicalin baicalein wogonoside wogo-nin berberine palmatine and jatrorrhizine were determinedin high fat-induced AS rats
International Journal of Analytical Chemistry 7
Baicali
n
Baicale
in
Wogonosid
e
Wogonin
Berberi
ne
Palmati
ne
Jatrorrh
izine
01
220
4020
0040
00C
once
ntra
tion
(ng
mL)
Low
Medium
High
Figure 3 The concentration of seven analytes in rat plasma after oral administration of three dosages of HLJDD
Table 2 Regression data and LLOQs of the multi-components determined in HLJDD
Although the determination of HLJDD was reportedbefore [31 32] the active components of it in biologicalsamples have seldom been reported Deng and He [33 34]determined baicalin and wogonoside in type 2 diabetic andnormal rats Zeng and Zhu [35 36] quantified baicalingeniposide and berberine in MCAO rats Nevertheless theywere all based on HPLCmethod and the LLOQwere 120583g levelfor those constituents Thus we quantified the above seveningredients simultaneously in the plasma of high fat-inducedAS rats after oral administration ofHLJDD except geniposide(for there was almost no geniposide in the drug-containingplasma and the pretreatment of geniposide was extremelyunstable) by UHPLC-MSMS In our study the precisionaccuracy matrix effect and stability under all conditionsare within bioanalytical methodology validation acceptancecriteria [37] with the extraction recovery of palmatine andberberine higher than Lursquos study [15] the LLOQ was lower
and the retention time greatly shortened than the previousresearch [33ndash36] Although some peaks eluted with closedretention time the MS detector can determine them accu-rately by taking the advantage of its high selective MRMmethod
43 Concentration Profiles of theAnalytes inASPlasma Fromthe result of determination of HLJDD-containing plasmabaicalin baicalein wogonoside andwogonin could be highlydetected in a dose-dependent manner while berberine jatr-orrhizine and palmatine were determined in a very lowlevel and in a dose-independent mode For one thing theformer absorptions were relatively better than the latterFor another flavonoids are easily bound to glucuronic acidor sulfuric acid to form two-phase metabolism so theirplasma concentration-time curves showed obvious bimodalphenomena and concentration increased slowly from the
8 International Journal of Analytical Chemistry
Table 3 Intra-inter-day precision and accuracy for the determination of the components in rat plasma
Analytes Spiked Concentration(ngmL)
Precision () Accuracy ( Mean plusmn SD)Intra-day Inter-day Intra-day Inter-day
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
SpectroscopyAnalytical ChemistryInternational Journal of
Hindawiwwwhindawicom Volume 2018
MaterialsJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
BioMed Research International Electrochemistry
International Journal of
Hindawiwwwhindawicom Volume 2018
Na
nom
ate
ria
ls
Hindawiwwwhindawicom Volume 2018
Journal ofNanomaterials
Submit your manuscripts atwwwhindawicom
International Journal of Analytical Chemistry 3
OOH
HO
O O
O
OHHOOC
OH
OH
Baicalin
OOH
HO
HO O
Baicalein
OOH
O O
Wogonoside
O
OH
HO
HO
COOHOOH
HO O
Wogonin
O
O
Berberine Palmatine
HO
Jatrorrhizine
O
O
O
O
OH
OH
OH
OH
OO
OH
OH
HO
Icariin
N
Tetrahydropalmatine
OCH3 OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
N+
H3CO
H3CO
H3CO
N+
H3C
HOH2C
N+
H3CO
H3CO
Figure 1 Chemical structures of baicalin baicalein wogonoside wogonin berberine palmatine jatrorrhizine icariin (IS) andtetrahydropalmatine (IS)
4 International Journal of Analytical Chemistry
250 and 575 ngml for baicalein 20 250 and 1875 ngmlfor wogonoside 20 257 and 13672 ngml for wogoninand 025 25 and 250 ngml for berberine palmatine andjatrorrhizine
25 Plasma Sample Preparation An aliquot of 100120583L thawedplasma sample was transferred into an Eppendorf tube (EPtube) to which 10 120583L of seven analytes and internal standardssolution from each working solution were added After beingvortexed for 15 s 100 120583L methanol plus 300 120583L acetonitrilewas added and rested for 3 h following vortex mixing for30 s to precipitate protein Subsequently the mixture wascentrifuged at 15000timesg for 10 min 400 120583L supernatant wastransferred into another EP tube and evaporated to drynessunder the stream of nitrogen in a water bath at 40∘C Theresidue was dissolved in 100 120583L of reconstituted solutionwhich consisted of acetonitrile-01 formic acid (91 vv) andthen centrifuged at 18000timesg for 15 min after vortexing for 2min The supernatant was injected into the UHPLCndashMSMSsystem for analysis
26 Instruments and Chromatographic Conditions The anal-ysis was performed using the Shimadzu UHPLC system(Shimadzu Corporation Kyoto Japan) consisting of an LC-30AD binary pump a DGU-20A5 degassing unit a SIL-30AC autosampler and a CTO-30A5R column oven Massspectrometric detectionwas conducted on anAB SciexQtrap5500 System (Applied Biosystems Foster City CA USA)and equipped with Analyst software (version 162) for dataprocessing Chromatographic separation was achieved ona Shimadzu Shim-pack XR-ODS III (16 120583m 20 mmtimes75mm ShimadzuCorporation Kyoto Japan)Themobile phaseconsisted of 5 mM ammonium formate-01 formic acid(solvent A) and acetonitrile (solvent B) The gradient elutionconditions were optimized as follows 10-20 B (0-3 min)20-45 B (3-45 min) 45 B (45-7 min) 45-80 B(7-12min) 80-95 B (12-14 min) 95-10B (15-155 min) and 10B (155-16 min) The flow rate was set at 03mLmin withthe column temperature maintained at 40∘C The injectionvolume of 1 120583L was used for the reference standards andsamples
For mass detection the electrospray ionization sourcewas operated in positive mode The operating parameterswere optimized under the following conditions 500∘C forthe interface temperature and 55 kV for the ion sprayvoltage ion source gas 1 and gas 2 were fixed at 50 psiVacuum was obtained by a Turbo molecular pump (AgilentTechnologies USA) Nitrogen generated by the high puritynitrogen generator (99999 Peak Scientific InstrumentsLtd UK) was used as the source of curtain gas (30 psi)and collision gas (7 psi) The optimized multiple reactionmonitoring (MRM) parameters including collision energyand declustering potential are listed in Table 1
27 Validation of the UPLCndashMSMS Method The proposedmethod was validated for specificity extraction recovery
matrix effect LLOQ linearity and stabilityMeanwhile intra-and interday validation were performed to evaluate theaccuracy and precision of the measurements
271 Specificity Linearity Lower Limits of Detection andQuantification To investigate the specificity of this methodchromatogram comparison of blank plasma blank plasmaspiked with ISanalyte and rat plasma samples were con-ducted to assay for the exclusion of any endogenous interfer-ence existing at or close to the expected retention time of theanalytes Calibration curves were established from peak arearatios (analyte peak area to the internal standard peak areaAsAi) versus nominal concentrations using a linear least-squares regression model (1X2 weighting)
272 Precision and Accuracy Intra- and interday precisionsand accuracy were denoted by assessing measured results ofQC samples at low medium and high concentrations Eachof the five samples was processed in parallel Continuouslymeasure a batch of samples and calculate intraday precisionThen five samples of low medium and high plasma samplesof each componentwere prepared in parallel on different con-secutive days and the 45 samples were tested in three batchesand the interday precision was calculated by referring to theaccompanying standard Precisions were expressed by therelative standard deviation (RSD ) while accuracy () waspresented as the percentage difference between the meanmeasured concentrations and the spiked concentrations
273 Extraction Recovery and Matrix Effect Extractionrecoveries of the seven analytes from rat blank plasmawere determined by comparing the mean peak areas of theQC samples spiked before protein precipitation with thosespiked after protein extraction Matrix effects occurred whenendogenous molecules were coeluting with the analytes ofinterest enhancing or decreasing the ionization efficiency ofthe electrospray interface The matrix effect was assessed viacomparing the mean peak areas of the QC samples spikedafter the pretreatment with those of the pure solution
274 Stability Stability of seven active constituents waschecked by comparing measured results with those of freshlyprepared samples The short- and long-term stabilities wereevaluated by analyzing QC plasma samples kept at roomtemperature for 4 h and in the freezer (minus20∘C) for 30 daysrespectively the freeze-thaw stability was carried out bydetecting QC samples undergoing three freeze-thaw cyclesthe postpreparation stability was assessed by determining theextracted QC samples stored under autosampler conditions(4∘C) for 12 h
28 Determination of HLJDD-Containing Plasma Specifi-cally pathogen-free Sprague-Dawley rats (male weighing200 plusmn 20 g) were purchased from Hunan Slac LaboratoryAnimal Co LTD (Hunan China Certificate No SCXK-2013-0004) and acclimated in Exhaust Ventilated Closed-SystemCage Rack (EVC) for at least a week with environmentallycontrolled quarters (22plusmn2∘C and 1212-h lightdark cycle) and
International Journal of Analytical Chemistry 5
Table 1 Optimized precursorproduction pairs and multiple reaction monitoring (MRM) parameters for the analytes and IS
free access to standard chow and water Animal welfare andexperimental procedures were strictly in accordance withthe guide for the care and use of laboratory animal by theAnimal Ethics Committee of Jiangxi University of TCMAfter one week of acclimatization the mice were randomlydivided into five groups (119899=12) normal control group modelgroup and three dosage groups The normal control groupwas fed with common diet while the other groups werefed with high-fat diet (3 cholesterol 05 sodium cholate02 propylthiouracil 5 sugar 10 lard and 813 basicdiet) The high fat-induced AS rat model was established inour previous study [17 18] Briefly the AS model was madeby the combination of regular intraperitoneal injection ofvitamin D
3and high fat diet for 8 weeks The model rat
was injected with 600000 UKg vitamin D3on the second
week and 200000 UKg every other week The three dosagegroups were gavaged HLJDD at doses of 15 3 and 6 g kg(low medium and high) once daily from the 3rd week for8 weeks until they were sacrificed Then the heparinizedblood samples were collected from abdominal aorta after 30minutes of the last administration and frozen at minus20∘C untilanalysis
3 Results
31 Method Validation
311 Specificity The total separation time for all analyteswas 16 minutes and there were little interferential substanceswith the analytes and IS in the blank plasma Representativechromatogram of analytes and IS in rat plasma was shown inFigure 2
312 Linearity The calibration curve of each analyte wasestablished with at least six points of standard solution andeach point was repeated five times The calibration curvesof all analytes exhibited good linearity and the regressionequations with correlation coefficients and linear range werelisted in Table 2
313 Precision and Accuracy The intraday and interdayprecision of all analytes were all less than 15 whilst theaccuracy deviation values were all within 964plusmn60 of theactual values at each QC level (shown in Table 3)
314 Extraction Recovery and Matrix Effect The extractionrecoveries (absolute recoveries) of each component weremore than 80 at each QC level which satisfied the quan-titative requirements of biological samples With respect tomatrix effect no suppressive or enhancing effect was foundon the analytes and IS That is to say the responses of allcomponents in the matrix were consistent with that in puresolution The results are shown in Table 4
315 Stability Results of the stability (shown in Table 5)illustrate that all analytes remained generally stable in plasmafor 4 h when stored at room temperature or 30 days whenstored atminus20∘C for three freeze-thaw cycles And they showedsatisfactory stability in the reconstituted solutions whenstored under autosampler condition for 12 h
316 Application The present method was successfully usedfor the determination of three dosages of HLJDD in 24 highfat-induced AS rat plasmasThe concentration of the analytesin plasma after ig administration was shown in Table 6 andFigure 3
4 Discussion
41 Optimization of LCndashMS for Quantitative Analysis Thechoice of mobile phase was a crucial factor in achievingfine chromatographic behavior and appropriate ionizationModifiers such as formic acid and ammonium formatealone or in combination with different concentrations werecomparedThe best peak shape and ionization were achievedadapting 5 mM ammonium formate buffer Linear gradientelution was used to elute endogenous substances residuefrom the column In addition all analytes and IS were bothfully scanned by positive and negative mode As alkaloidswere detected overwhelmingly in the positive mode whileflavonoids with little difference in both modes the positivemode was used in the MRM acquisition
42 Selection of the Determined Components According tothe previous phytochemical and HPLCndashMS studies iridoidsalkaloids and flavonoids were the predominant constituentsin HLJDD [19ndash23] And many components of HLJDD could
6 International Journal of Analytical Chemistry
2 4 6 80
110
2 4 6 80
250
2 4 6 80
180
2 4 6 80
150
palmatine
jateorhizine
icariin
tetrahydropalmatine
2 4 6 80
85
2 4 6 80
225
2 4 6 80
275
2 4 6 80
300
2 4 6 80
350
baicalin
baicalein
wogonoside
wogonin
berberine
Inte
nsity
(CPS
)
Time (min)
(a)
2 4 6 80
12000
2 4 6 80
70000
2 4 6 80
2750
2 4 6 80
18000
54
58
54
56
2 4 6 80
14000
2 4 6 80
4500
2 4 6 80
40000
2 4 6 80
50000
2 4 6 80
650000
58
59
58
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(b)
2 4 6 80
4250
2 4 6 80
1100
2 4 6 80
850000
2 4 6 80
1700000
56
54
58
54
2 4 6 80
110000
2 4 6 80
10000
2 4 6 80
750000
2 4 6 80
22500
2 4 6 80
6500
58
58
59
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(c)
Figure 2 The MRM spectrum of each component (a) blank plasma (b) blank plasma spiked with analytes and IS (c) rat plasma samplecollected after ig administration of HLJDD
alleviate AS development by inhibiting the vascular inflam-matory processes which was the initiation and progressionof AS [3 4] The flavonoids of Radix Scutellariae iebaicalin baicalein and wogonin could suppress vascularinflammation in vitro and in vivo [24] andwogonoside couldmodulate inflammatory mediator expression in LPS-inducedRAW2647 cells [25] In addition wogonin inhibited MMP-9 gene expression (a major role in the pathogenesis of AS)
via MAPK signaling pathways [26] Berberine palmatineand jatrorrhizine the main alkaloids constituents in Rhi-zome Coptidis and Cortex Phellodendri could suppress theformation and development of AS by altering gut microbiotacompositions [27] anti-inflammation and lowering bloodlipids [28ndash30] Thus baicalin baicalein wogonoside wogo-nin berberine palmatine and jatrorrhizine were determinedin high fat-induced AS rats
International Journal of Analytical Chemistry 7
Baicali
n
Baicale
in
Wogonosid
e
Wogonin
Berberi
ne
Palmati
ne
Jatrorrh
izine
01
220
4020
0040
00C
once
ntra
tion
(ng
mL)
Low
Medium
High
Figure 3 The concentration of seven analytes in rat plasma after oral administration of three dosages of HLJDD
Table 2 Regression data and LLOQs of the multi-components determined in HLJDD
Although the determination of HLJDD was reportedbefore [31 32] the active components of it in biologicalsamples have seldom been reported Deng and He [33 34]determined baicalin and wogonoside in type 2 diabetic andnormal rats Zeng and Zhu [35 36] quantified baicalingeniposide and berberine in MCAO rats Nevertheless theywere all based on HPLCmethod and the LLOQwere 120583g levelfor those constituents Thus we quantified the above seveningredients simultaneously in the plasma of high fat-inducedAS rats after oral administration ofHLJDD except geniposide(for there was almost no geniposide in the drug-containingplasma and the pretreatment of geniposide was extremelyunstable) by UHPLC-MSMS In our study the precisionaccuracy matrix effect and stability under all conditionsare within bioanalytical methodology validation acceptancecriteria [37] with the extraction recovery of palmatine andberberine higher than Lursquos study [15] the LLOQ was lower
and the retention time greatly shortened than the previousresearch [33ndash36] Although some peaks eluted with closedretention time the MS detector can determine them accu-rately by taking the advantage of its high selective MRMmethod
43 Concentration Profiles of theAnalytes inASPlasma Fromthe result of determination of HLJDD-containing plasmabaicalin baicalein wogonoside andwogonin could be highlydetected in a dose-dependent manner while berberine jatr-orrhizine and palmatine were determined in a very lowlevel and in a dose-independent mode For one thing theformer absorptions were relatively better than the latterFor another flavonoids are easily bound to glucuronic acidor sulfuric acid to form two-phase metabolism so theirplasma concentration-time curves showed obvious bimodalphenomena and concentration increased slowly from the
8 International Journal of Analytical Chemistry
Table 3 Intra-inter-day precision and accuracy for the determination of the components in rat plasma
Analytes Spiked Concentration(ngmL)
Precision () Accuracy ( Mean plusmn SD)Intra-day Inter-day Intra-day Inter-day
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
SpectroscopyAnalytical ChemistryInternational Journal of
Hindawiwwwhindawicom Volume 2018
MaterialsJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
BioMed Research International Electrochemistry
International Journal of
Hindawiwwwhindawicom Volume 2018
Na
nom
ate
ria
ls
Hindawiwwwhindawicom Volume 2018
Journal ofNanomaterials
Submit your manuscripts atwwwhindawicom
4 International Journal of Analytical Chemistry
250 and 575 ngml for baicalein 20 250 and 1875 ngmlfor wogonoside 20 257 and 13672 ngml for wogoninand 025 25 and 250 ngml for berberine palmatine andjatrorrhizine
25 Plasma Sample Preparation An aliquot of 100120583L thawedplasma sample was transferred into an Eppendorf tube (EPtube) to which 10 120583L of seven analytes and internal standardssolution from each working solution were added After beingvortexed for 15 s 100 120583L methanol plus 300 120583L acetonitrilewas added and rested for 3 h following vortex mixing for30 s to precipitate protein Subsequently the mixture wascentrifuged at 15000timesg for 10 min 400 120583L supernatant wastransferred into another EP tube and evaporated to drynessunder the stream of nitrogen in a water bath at 40∘C Theresidue was dissolved in 100 120583L of reconstituted solutionwhich consisted of acetonitrile-01 formic acid (91 vv) andthen centrifuged at 18000timesg for 15 min after vortexing for 2min The supernatant was injected into the UHPLCndashMSMSsystem for analysis
26 Instruments and Chromatographic Conditions The anal-ysis was performed using the Shimadzu UHPLC system(Shimadzu Corporation Kyoto Japan) consisting of an LC-30AD binary pump a DGU-20A5 degassing unit a SIL-30AC autosampler and a CTO-30A5R column oven Massspectrometric detectionwas conducted on anAB SciexQtrap5500 System (Applied Biosystems Foster City CA USA)and equipped with Analyst software (version 162) for dataprocessing Chromatographic separation was achieved ona Shimadzu Shim-pack XR-ODS III (16 120583m 20 mmtimes75mm ShimadzuCorporation Kyoto Japan)Themobile phaseconsisted of 5 mM ammonium formate-01 formic acid(solvent A) and acetonitrile (solvent B) The gradient elutionconditions were optimized as follows 10-20 B (0-3 min)20-45 B (3-45 min) 45 B (45-7 min) 45-80 B(7-12min) 80-95 B (12-14 min) 95-10B (15-155 min) and 10B (155-16 min) The flow rate was set at 03mLmin withthe column temperature maintained at 40∘C The injectionvolume of 1 120583L was used for the reference standards andsamples
For mass detection the electrospray ionization sourcewas operated in positive mode The operating parameterswere optimized under the following conditions 500∘C forthe interface temperature and 55 kV for the ion sprayvoltage ion source gas 1 and gas 2 were fixed at 50 psiVacuum was obtained by a Turbo molecular pump (AgilentTechnologies USA) Nitrogen generated by the high puritynitrogen generator (99999 Peak Scientific InstrumentsLtd UK) was used as the source of curtain gas (30 psi)and collision gas (7 psi) The optimized multiple reactionmonitoring (MRM) parameters including collision energyand declustering potential are listed in Table 1
27 Validation of the UPLCndashMSMS Method The proposedmethod was validated for specificity extraction recovery
matrix effect LLOQ linearity and stabilityMeanwhile intra-and interday validation were performed to evaluate theaccuracy and precision of the measurements
271 Specificity Linearity Lower Limits of Detection andQuantification To investigate the specificity of this methodchromatogram comparison of blank plasma blank plasmaspiked with ISanalyte and rat plasma samples were con-ducted to assay for the exclusion of any endogenous interfer-ence existing at or close to the expected retention time of theanalytes Calibration curves were established from peak arearatios (analyte peak area to the internal standard peak areaAsAi) versus nominal concentrations using a linear least-squares regression model (1X2 weighting)
272 Precision and Accuracy Intra- and interday precisionsand accuracy were denoted by assessing measured results ofQC samples at low medium and high concentrations Eachof the five samples was processed in parallel Continuouslymeasure a batch of samples and calculate intraday precisionThen five samples of low medium and high plasma samplesof each componentwere prepared in parallel on different con-secutive days and the 45 samples were tested in three batchesand the interday precision was calculated by referring to theaccompanying standard Precisions were expressed by therelative standard deviation (RSD ) while accuracy () waspresented as the percentage difference between the meanmeasured concentrations and the spiked concentrations
273 Extraction Recovery and Matrix Effect Extractionrecoveries of the seven analytes from rat blank plasmawere determined by comparing the mean peak areas of theQC samples spiked before protein precipitation with thosespiked after protein extraction Matrix effects occurred whenendogenous molecules were coeluting with the analytes ofinterest enhancing or decreasing the ionization efficiency ofthe electrospray interface The matrix effect was assessed viacomparing the mean peak areas of the QC samples spikedafter the pretreatment with those of the pure solution
274 Stability Stability of seven active constituents waschecked by comparing measured results with those of freshlyprepared samples The short- and long-term stabilities wereevaluated by analyzing QC plasma samples kept at roomtemperature for 4 h and in the freezer (minus20∘C) for 30 daysrespectively the freeze-thaw stability was carried out bydetecting QC samples undergoing three freeze-thaw cyclesthe postpreparation stability was assessed by determining theextracted QC samples stored under autosampler conditions(4∘C) for 12 h
28 Determination of HLJDD-Containing Plasma Specifi-cally pathogen-free Sprague-Dawley rats (male weighing200 plusmn 20 g) were purchased from Hunan Slac LaboratoryAnimal Co LTD (Hunan China Certificate No SCXK-2013-0004) and acclimated in Exhaust Ventilated Closed-SystemCage Rack (EVC) for at least a week with environmentallycontrolled quarters (22plusmn2∘C and 1212-h lightdark cycle) and
International Journal of Analytical Chemistry 5
Table 1 Optimized precursorproduction pairs and multiple reaction monitoring (MRM) parameters for the analytes and IS
free access to standard chow and water Animal welfare andexperimental procedures were strictly in accordance withthe guide for the care and use of laboratory animal by theAnimal Ethics Committee of Jiangxi University of TCMAfter one week of acclimatization the mice were randomlydivided into five groups (119899=12) normal control group modelgroup and three dosage groups The normal control groupwas fed with common diet while the other groups werefed with high-fat diet (3 cholesterol 05 sodium cholate02 propylthiouracil 5 sugar 10 lard and 813 basicdiet) The high fat-induced AS rat model was established inour previous study [17 18] Briefly the AS model was madeby the combination of regular intraperitoneal injection ofvitamin D
3and high fat diet for 8 weeks The model rat
was injected with 600000 UKg vitamin D3on the second
week and 200000 UKg every other week The three dosagegroups were gavaged HLJDD at doses of 15 3 and 6 g kg(low medium and high) once daily from the 3rd week for8 weeks until they were sacrificed Then the heparinizedblood samples were collected from abdominal aorta after 30minutes of the last administration and frozen at minus20∘C untilanalysis
3 Results
31 Method Validation
311 Specificity The total separation time for all analyteswas 16 minutes and there were little interferential substanceswith the analytes and IS in the blank plasma Representativechromatogram of analytes and IS in rat plasma was shown inFigure 2
312 Linearity The calibration curve of each analyte wasestablished with at least six points of standard solution andeach point was repeated five times The calibration curvesof all analytes exhibited good linearity and the regressionequations with correlation coefficients and linear range werelisted in Table 2
313 Precision and Accuracy The intraday and interdayprecision of all analytes were all less than 15 whilst theaccuracy deviation values were all within 964plusmn60 of theactual values at each QC level (shown in Table 3)
314 Extraction Recovery and Matrix Effect The extractionrecoveries (absolute recoveries) of each component weremore than 80 at each QC level which satisfied the quan-titative requirements of biological samples With respect tomatrix effect no suppressive or enhancing effect was foundon the analytes and IS That is to say the responses of allcomponents in the matrix were consistent with that in puresolution The results are shown in Table 4
315 Stability Results of the stability (shown in Table 5)illustrate that all analytes remained generally stable in plasmafor 4 h when stored at room temperature or 30 days whenstored atminus20∘C for three freeze-thaw cycles And they showedsatisfactory stability in the reconstituted solutions whenstored under autosampler condition for 12 h
316 Application The present method was successfully usedfor the determination of three dosages of HLJDD in 24 highfat-induced AS rat plasmasThe concentration of the analytesin plasma after ig administration was shown in Table 6 andFigure 3
4 Discussion
41 Optimization of LCndashMS for Quantitative Analysis Thechoice of mobile phase was a crucial factor in achievingfine chromatographic behavior and appropriate ionizationModifiers such as formic acid and ammonium formatealone or in combination with different concentrations werecomparedThe best peak shape and ionization were achievedadapting 5 mM ammonium formate buffer Linear gradientelution was used to elute endogenous substances residuefrom the column In addition all analytes and IS were bothfully scanned by positive and negative mode As alkaloidswere detected overwhelmingly in the positive mode whileflavonoids with little difference in both modes the positivemode was used in the MRM acquisition
42 Selection of the Determined Components According tothe previous phytochemical and HPLCndashMS studies iridoidsalkaloids and flavonoids were the predominant constituentsin HLJDD [19ndash23] And many components of HLJDD could
6 International Journal of Analytical Chemistry
2 4 6 80
110
2 4 6 80
250
2 4 6 80
180
2 4 6 80
150
palmatine
jateorhizine
icariin
tetrahydropalmatine
2 4 6 80
85
2 4 6 80
225
2 4 6 80
275
2 4 6 80
300
2 4 6 80
350
baicalin
baicalein
wogonoside
wogonin
berberine
Inte
nsity
(CPS
)
Time (min)
(a)
2 4 6 80
12000
2 4 6 80
70000
2 4 6 80
2750
2 4 6 80
18000
54
58
54
56
2 4 6 80
14000
2 4 6 80
4500
2 4 6 80
40000
2 4 6 80
50000
2 4 6 80
650000
58
59
58
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(b)
2 4 6 80
4250
2 4 6 80
1100
2 4 6 80
850000
2 4 6 80
1700000
56
54
58
54
2 4 6 80
110000
2 4 6 80
10000
2 4 6 80
750000
2 4 6 80
22500
2 4 6 80
6500
58
58
59
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(c)
Figure 2 The MRM spectrum of each component (a) blank plasma (b) blank plasma spiked with analytes and IS (c) rat plasma samplecollected after ig administration of HLJDD
alleviate AS development by inhibiting the vascular inflam-matory processes which was the initiation and progressionof AS [3 4] The flavonoids of Radix Scutellariae iebaicalin baicalein and wogonin could suppress vascularinflammation in vitro and in vivo [24] andwogonoside couldmodulate inflammatory mediator expression in LPS-inducedRAW2647 cells [25] In addition wogonin inhibited MMP-9 gene expression (a major role in the pathogenesis of AS)
via MAPK signaling pathways [26] Berberine palmatineand jatrorrhizine the main alkaloids constituents in Rhi-zome Coptidis and Cortex Phellodendri could suppress theformation and development of AS by altering gut microbiotacompositions [27] anti-inflammation and lowering bloodlipids [28ndash30] Thus baicalin baicalein wogonoside wogo-nin berberine palmatine and jatrorrhizine were determinedin high fat-induced AS rats
International Journal of Analytical Chemistry 7
Baicali
n
Baicale
in
Wogonosid
e
Wogonin
Berberi
ne
Palmati
ne
Jatrorrh
izine
01
220
4020
0040
00C
once
ntra
tion
(ng
mL)
Low
Medium
High
Figure 3 The concentration of seven analytes in rat plasma after oral administration of three dosages of HLJDD
Table 2 Regression data and LLOQs of the multi-components determined in HLJDD
Although the determination of HLJDD was reportedbefore [31 32] the active components of it in biologicalsamples have seldom been reported Deng and He [33 34]determined baicalin and wogonoside in type 2 diabetic andnormal rats Zeng and Zhu [35 36] quantified baicalingeniposide and berberine in MCAO rats Nevertheless theywere all based on HPLCmethod and the LLOQwere 120583g levelfor those constituents Thus we quantified the above seveningredients simultaneously in the plasma of high fat-inducedAS rats after oral administration ofHLJDD except geniposide(for there was almost no geniposide in the drug-containingplasma and the pretreatment of geniposide was extremelyunstable) by UHPLC-MSMS In our study the precisionaccuracy matrix effect and stability under all conditionsare within bioanalytical methodology validation acceptancecriteria [37] with the extraction recovery of palmatine andberberine higher than Lursquos study [15] the LLOQ was lower
and the retention time greatly shortened than the previousresearch [33ndash36] Although some peaks eluted with closedretention time the MS detector can determine them accu-rately by taking the advantage of its high selective MRMmethod
43 Concentration Profiles of theAnalytes inASPlasma Fromthe result of determination of HLJDD-containing plasmabaicalin baicalein wogonoside andwogonin could be highlydetected in a dose-dependent manner while berberine jatr-orrhizine and palmatine were determined in a very lowlevel and in a dose-independent mode For one thing theformer absorptions were relatively better than the latterFor another flavonoids are easily bound to glucuronic acidor sulfuric acid to form two-phase metabolism so theirplasma concentration-time curves showed obvious bimodalphenomena and concentration increased slowly from the
8 International Journal of Analytical Chemistry
Table 3 Intra-inter-day precision and accuracy for the determination of the components in rat plasma
Analytes Spiked Concentration(ngmL)
Precision () Accuracy ( Mean plusmn SD)Intra-day Inter-day Intra-day Inter-day
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
free access to standard chow and water Animal welfare andexperimental procedures were strictly in accordance withthe guide for the care and use of laboratory animal by theAnimal Ethics Committee of Jiangxi University of TCMAfter one week of acclimatization the mice were randomlydivided into five groups (119899=12) normal control group modelgroup and three dosage groups The normal control groupwas fed with common diet while the other groups werefed with high-fat diet (3 cholesterol 05 sodium cholate02 propylthiouracil 5 sugar 10 lard and 813 basicdiet) The high fat-induced AS rat model was established inour previous study [17 18] Briefly the AS model was madeby the combination of regular intraperitoneal injection ofvitamin D
3and high fat diet for 8 weeks The model rat
was injected with 600000 UKg vitamin D3on the second
week and 200000 UKg every other week The three dosagegroups were gavaged HLJDD at doses of 15 3 and 6 g kg(low medium and high) once daily from the 3rd week for8 weeks until they were sacrificed Then the heparinizedblood samples were collected from abdominal aorta after 30minutes of the last administration and frozen at minus20∘C untilanalysis
3 Results
31 Method Validation
311 Specificity The total separation time for all analyteswas 16 minutes and there were little interferential substanceswith the analytes and IS in the blank plasma Representativechromatogram of analytes and IS in rat plasma was shown inFigure 2
312 Linearity The calibration curve of each analyte wasestablished with at least six points of standard solution andeach point was repeated five times The calibration curvesof all analytes exhibited good linearity and the regressionequations with correlation coefficients and linear range werelisted in Table 2
313 Precision and Accuracy The intraday and interdayprecision of all analytes were all less than 15 whilst theaccuracy deviation values were all within 964plusmn60 of theactual values at each QC level (shown in Table 3)
314 Extraction Recovery and Matrix Effect The extractionrecoveries (absolute recoveries) of each component weremore than 80 at each QC level which satisfied the quan-titative requirements of biological samples With respect tomatrix effect no suppressive or enhancing effect was foundon the analytes and IS That is to say the responses of allcomponents in the matrix were consistent with that in puresolution The results are shown in Table 4
315 Stability Results of the stability (shown in Table 5)illustrate that all analytes remained generally stable in plasmafor 4 h when stored at room temperature or 30 days whenstored atminus20∘C for three freeze-thaw cycles And they showedsatisfactory stability in the reconstituted solutions whenstored under autosampler condition for 12 h
316 Application The present method was successfully usedfor the determination of three dosages of HLJDD in 24 highfat-induced AS rat plasmasThe concentration of the analytesin plasma after ig administration was shown in Table 6 andFigure 3
4 Discussion
41 Optimization of LCndashMS for Quantitative Analysis Thechoice of mobile phase was a crucial factor in achievingfine chromatographic behavior and appropriate ionizationModifiers such as formic acid and ammonium formatealone or in combination with different concentrations werecomparedThe best peak shape and ionization were achievedadapting 5 mM ammonium formate buffer Linear gradientelution was used to elute endogenous substances residuefrom the column In addition all analytes and IS were bothfully scanned by positive and negative mode As alkaloidswere detected overwhelmingly in the positive mode whileflavonoids with little difference in both modes the positivemode was used in the MRM acquisition
42 Selection of the Determined Components According tothe previous phytochemical and HPLCndashMS studies iridoidsalkaloids and flavonoids were the predominant constituentsin HLJDD [19ndash23] And many components of HLJDD could
6 International Journal of Analytical Chemistry
2 4 6 80
110
2 4 6 80
250
2 4 6 80
180
2 4 6 80
150
palmatine
jateorhizine
icariin
tetrahydropalmatine
2 4 6 80
85
2 4 6 80
225
2 4 6 80
275
2 4 6 80
300
2 4 6 80
350
baicalin
baicalein
wogonoside
wogonin
berberine
Inte
nsity
(CPS
)
Time (min)
(a)
2 4 6 80
12000
2 4 6 80
70000
2 4 6 80
2750
2 4 6 80
18000
54
58
54
56
2 4 6 80
14000
2 4 6 80
4500
2 4 6 80
40000
2 4 6 80
50000
2 4 6 80
650000
58
59
58
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(b)
2 4 6 80
4250
2 4 6 80
1100
2 4 6 80
850000
2 4 6 80
1700000
56
54
58
54
2 4 6 80
110000
2 4 6 80
10000
2 4 6 80
750000
2 4 6 80
22500
2 4 6 80
6500
58
58
59
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(c)
Figure 2 The MRM spectrum of each component (a) blank plasma (b) blank plasma spiked with analytes and IS (c) rat plasma samplecollected after ig administration of HLJDD
alleviate AS development by inhibiting the vascular inflam-matory processes which was the initiation and progressionof AS [3 4] The flavonoids of Radix Scutellariae iebaicalin baicalein and wogonin could suppress vascularinflammation in vitro and in vivo [24] andwogonoside couldmodulate inflammatory mediator expression in LPS-inducedRAW2647 cells [25] In addition wogonin inhibited MMP-9 gene expression (a major role in the pathogenesis of AS)
via MAPK signaling pathways [26] Berberine palmatineand jatrorrhizine the main alkaloids constituents in Rhi-zome Coptidis and Cortex Phellodendri could suppress theformation and development of AS by altering gut microbiotacompositions [27] anti-inflammation and lowering bloodlipids [28ndash30] Thus baicalin baicalein wogonoside wogo-nin berberine palmatine and jatrorrhizine were determinedin high fat-induced AS rats
International Journal of Analytical Chemistry 7
Baicali
n
Baicale
in
Wogonosid
e
Wogonin
Berberi
ne
Palmati
ne
Jatrorrh
izine
01
220
4020
0040
00C
once
ntra
tion
(ng
mL)
Low
Medium
High
Figure 3 The concentration of seven analytes in rat plasma after oral administration of three dosages of HLJDD
Table 2 Regression data and LLOQs of the multi-components determined in HLJDD
Although the determination of HLJDD was reportedbefore [31 32] the active components of it in biologicalsamples have seldom been reported Deng and He [33 34]determined baicalin and wogonoside in type 2 diabetic andnormal rats Zeng and Zhu [35 36] quantified baicalingeniposide and berberine in MCAO rats Nevertheless theywere all based on HPLCmethod and the LLOQwere 120583g levelfor those constituents Thus we quantified the above seveningredients simultaneously in the plasma of high fat-inducedAS rats after oral administration ofHLJDD except geniposide(for there was almost no geniposide in the drug-containingplasma and the pretreatment of geniposide was extremelyunstable) by UHPLC-MSMS In our study the precisionaccuracy matrix effect and stability under all conditionsare within bioanalytical methodology validation acceptancecriteria [37] with the extraction recovery of palmatine andberberine higher than Lursquos study [15] the LLOQ was lower
and the retention time greatly shortened than the previousresearch [33ndash36] Although some peaks eluted with closedretention time the MS detector can determine them accu-rately by taking the advantage of its high selective MRMmethod
43 Concentration Profiles of theAnalytes inASPlasma Fromthe result of determination of HLJDD-containing plasmabaicalin baicalein wogonoside andwogonin could be highlydetected in a dose-dependent manner while berberine jatr-orrhizine and palmatine were determined in a very lowlevel and in a dose-independent mode For one thing theformer absorptions were relatively better than the latterFor another flavonoids are easily bound to glucuronic acidor sulfuric acid to form two-phase metabolism so theirplasma concentration-time curves showed obvious bimodalphenomena and concentration increased slowly from the
8 International Journal of Analytical Chemistry
Table 3 Intra-inter-day precision and accuracy for the determination of the components in rat plasma
Analytes Spiked Concentration(ngmL)
Precision () Accuracy ( Mean plusmn SD)Intra-day Inter-day Intra-day Inter-day
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
SpectroscopyAnalytical ChemistryInternational Journal of
Hindawiwwwhindawicom Volume 2018
MaterialsJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
BioMed Research International Electrochemistry
International Journal of
Hindawiwwwhindawicom Volume 2018
Na
nom
ate
ria
ls
Hindawiwwwhindawicom Volume 2018
Journal ofNanomaterials
Submit your manuscripts atwwwhindawicom
6 International Journal of Analytical Chemistry
2 4 6 80
110
2 4 6 80
250
2 4 6 80
180
2 4 6 80
150
palmatine
jateorhizine
icariin
tetrahydropalmatine
2 4 6 80
85
2 4 6 80
225
2 4 6 80
275
2 4 6 80
300
2 4 6 80
350
baicalin
baicalein
wogonoside
wogonin
berberine
Inte
nsity
(CPS
)
Time (min)
(a)
2 4 6 80
12000
2 4 6 80
70000
2 4 6 80
2750
2 4 6 80
18000
54
58
54
56
2 4 6 80
14000
2 4 6 80
4500
2 4 6 80
40000
2 4 6 80
50000
2 4 6 80
650000
58
59
58
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(b)
2 4 6 80
4250
2 4 6 80
1100
2 4 6 80
850000
2 4 6 80
1700000
56
54
58
54
2 4 6 80
110000
2 4 6 80
10000
2 4 6 80
750000
2 4 6 80
22500
2 4 6 80
6500
58
58
59
58
54
Inte
nsity
(CPS
)
Time (min)
palmatine
jateorhizine
icariin
tetrahydropalmatine
baicalin
baicalein
wogonoside
wogonin
berberine
(c)
Figure 2 The MRM spectrum of each component (a) blank plasma (b) blank plasma spiked with analytes and IS (c) rat plasma samplecollected after ig administration of HLJDD
alleviate AS development by inhibiting the vascular inflam-matory processes which was the initiation and progressionof AS [3 4] The flavonoids of Radix Scutellariae iebaicalin baicalein and wogonin could suppress vascularinflammation in vitro and in vivo [24] andwogonoside couldmodulate inflammatory mediator expression in LPS-inducedRAW2647 cells [25] In addition wogonin inhibited MMP-9 gene expression (a major role in the pathogenesis of AS)
via MAPK signaling pathways [26] Berberine palmatineand jatrorrhizine the main alkaloids constituents in Rhi-zome Coptidis and Cortex Phellodendri could suppress theformation and development of AS by altering gut microbiotacompositions [27] anti-inflammation and lowering bloodlipids [28ndash30] Thus baicalin baicalein wogonoside wogo-nin berberine palmatine and jatrorrhizine were determinedin high fat-induced AS rats
International Journal of Analytical Chemistry 7
Baicali
n
Baicale
in
Wogonosid
e
Wogonin
Berberi
ne
Palmati
ne
Jatrorrh
izine
01
220
4020
0040
00C
once
ntra
tion
(ng
mL)
Low
Medium
High
Figure 3 The concentration of seven analytes in rat plasma after oral administration of three dosages of HLJDD
Table 2 Regression data and LLOQs of the multi-components determined in HLJDD
Although the determination of HLJDD was reportedbefore [31 32] the active components of it in biologicalsamples have seldom been reported Deng and He [33 34]determined baicalin and wogonoside in type 2 diabetic andnormal rats Zeng and Zhu [35 36] quantified baicalingeniposide and berberine in MCAO rats Nevertheless theywere all based on HPLCmethod and the LLOQwere 120583g levelfor those constituents Thus we quantified the above seveningredients simultaneously in the plasma of high fat-inducedAS rats after oral administration ofHLJDD except geniposide(for there was almost no geniposide in the drug-containingplasma and the pretreatment of geniposide was extremelyunstable) by UHPLC-MSMS In our study the precisionaccuracy matrix effect and stability under all conditionsare within bioanalytical methodology validation acceptancecriteria [37] with the extraction recovery of palmatine andberberine higher than Lursquos study [15] the LLOQ was lower
and the retention time greatly shortened than the previousresearch [33ndash36] Although some peaks eluted with closedretention time the MS detector can determine them accu-rately by taking the advantage of its high selective MRMmethod
43 Concentration Profiles of theAnalytes inASPlasma Fromthe result of determination of HLJDD-containing plasmabaicalin baicalein wogonoside andwogonin could be highlydetected in a dose-dependent manner while berberine jatr-orrhizine and palmatine were determined in a very lowlevel and in a dose-independent mode For one thing theformer absorptions were relatively better than the latterFor another flavonoids are easily bound to glucuronic acidor sulfuric acid to form two-phase metabolism so theirplasma concentration-time curves showed obvious bimodalphenomena and concentration increased slowly from the
8 International Journal of Analytical Chemistry
Table 3 Intra-inter-day precision and accuracy for the determination of the components in rat plasma
Analytes Spiked Concentration(ngmL)
Precision () Accuracy ( Mean plusmn SD)Intra-day Inter-day Intra-day Inter-day
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
Although the determination of HLJDD was reportedbefore [31 32] the active components of it in biologicalsamples have seldom been reported Deng and He [33 34]determined baicalin and wogonoside in type 2 diabetic andnormal rats Zeng and Zhu [35 36] quantified baicalingeniposide and berberine in MCAO rats Nevertheless theywere all based on HPLCmethod and the LLOQwere 120583g levelfor those constituents Thus we quantified the above seveningredients simultaneously in the plasma of high fat-inducedAS rats after oral administration ofHLJDD except geniposide(for there was almost no geniposide in the drug-containingplasma and the pretreatment of geniposide was extremelyunstable) by UHPLC-MSMS In our study the precisionaccuracy matrix effect and stability under all conditionsare within bioanalytical methodology validation acceptancecriteria [37] with the extraction recovery of palmatine andberberine higher than Lursquos study [15] the LLOQ was lower
and the retention time greatly shortened than the previousresearch [33ndash36] Although some peaks eluted with closedretention time the MS detector can determine them accu-rately by taking the advantage of its high selective MRMmethod
43 Concentration Profiles of theAnalytes inASPlasma Fromthe result of determination of HLJDD-containing plasmabaicalin baicalein wogonoside andwogonin could be highlydetected in a dose-dependent manner while berberine jatr-orrhizine and palmatine were determined in a very lowlevel and in a dose-independent mode For one thing theformer absorptions were relatively better than the latterFor another flavonoids are easily bound to glucuronic acidor sulfuric acid to form two-phase metabolism so theirplasma concentration-time curves showed obvious bimodalphenomena and concentration increased slowly from the
8 International Journal of Analytical Chemistry
Table 3 Intra-inter-day precision and accuracy for the determination of the components in rat plasma
Analytes Spiked Concentration(ngmL)
Precision () Accuracy ( Mean plusmn SD)Intra-day Inter-day Intra-day Inter-day
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
SpectroscopyAnalytical ChemistryInternational Journal of
Hindawiwwwhindawicom Volume 2018
MaterialsJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
BioMed Research International Electrochemistry
International Journal of
Hindawiwwwhindawicom Volume 2018
Na
nom
ate
ria
ls
Hindawiwwwhindawicom Volume 2018
Journal ofNanomaterials
Submit your manuscripts atwwwhindawicom
10 International Journal of Analytical Chemistry
5th day [38] However even long-term administration ofberberine and other alkaloids was not easy to accumulatein vivo for their poor absorption through the gut wallFirst of all berberine had strong rigidity and poor solubilityfor it is a quaternary ammonium alkaloid with conjugateddouble bonds Besides berberine was the substrate of P-gp which is an efflux transporter All these factors lead tothe poor absorption of berberine Secondly most of themwere excluded by the gastrointestinal tract after intragastricadministration of berberine and they were also metabolizedthrough various other pathways at the same time [39 40]Moreover the distribution of berberine in the organs wasmuch higher than that in the blood such as liver kidneyand muscle [41] Pharmacokinetic studies [15 39] indicatedthe blood clearance of berberine was very fast and itsbiotransformation in the liver was rapid and substantial Sothe first pass elimination of intestinal tract and the tendencydistribution of liver could also lead to the low concentrationof berberine in blood Furthermore our previous studies [42]have shown that baicalin is a partial agonist of berberinewhich weakened the pharmacological effect of berberine ina higher concentration range Therefore it may contribute tothe low blood concentration of berberine in vivo Likewisethe structures of jatrorrhizine and palmatine are similar tothat of berberine so the low blood concentration of thesetwo alkaloids may also be caused by similar reasons withberberine for the principle of structural similarity
As we all know the absorption of the intestinal tract andmetabolism of the liver may affect the bioavailability of thedrug [43] and the pathological conditions ie AS may alsoaffect the process of the drug in the body In humans thedevelopment of metabolic diseases including AS has closelyrelated to imbalance intestinal flora [44] and the intestinalflora may also affect the process of drugs in vivo So the effectof AS on the above constituents cannot be ignored
5 Conclusions
Quantification of ingredients at a low level was the obstaclesin the study of active components of traditional Chinesemedicine in biological fluids Simply using chromatographywas usually time-consuming insensitive and nonselectiveenough In the present study a highly selective and sensitiveUHPLCndashESI-MS method was developed and validated tosimultaneously determine the seven active components inrat plasma and successfully applied to 24 high fat-inducedAS rats after oral administration of HLJDD It could applyfor further pharmacokinetic study of the analytes and mayprovide a scientific basis for clinical application of HLJDD
Data Availability
The data used to support the findings of this study areavailable from the corresponding author upon request
Conflicts of Interest
The authors declared no conflicts of interest
Authorsrsquo Contributions
Li Jiang and Yanling Xiong contributed equally to this work
Acknowledgments
This work was supported by grants of National NaturalScience Foundation of China (Nos 81703823 and 81560744)Jiangxi Provincial Medical and Health Science amp TechnologyPlan (No 2018B131) Jiangxi Provincial Educational Scienceamp Technology Plan (No GJJ170753) and Jiangxi ProvincialChinese Medicine First Class Discipline Research Fund(JXSYLXK-ZHYAO120)
References
[1] L G Spagnoli E Bonanno G Sangiorgi and A MaurielloldquoRole of inflammation in atherosclerosisrdquo Journal of NuclearMedicine vol 48 no 11 pp 1800ndash1815 2007
[2] C J Wang J T Liu F Guo Y Ji and N Liu ldquoEndothelin-1 induces the expression of C-reactive protein in rat vascularsmooth muscle cellsrdquo Biochemical amp Biophysical Research Com-munications vol 389 pp 537ndash542 2009
[3] R Ross ldquoAtherosclerosisndashan inflammatory diseaserdquo The NewEngland Journal of Medicine vol 340 no 2 pp 115ndash126 1999
[4] K M Qin H Guo Z S Xu Z Q Yao and B C Cai ldquoResearchstatus of chemical constituents and pharmacokinetics of huan-glian jiedu tangrdquo Anti-Infection Pharm vol 8 pp 3ndash7 2011
[5] W P Wang ldquoEffect of basic Therapy combined with Huanglianjiedu decoction on carotid Atherosclerotic plaque and inflam-matory factors in patients with Coronary Heart Diseaserdquo Chi-nese Journal of Integrative Medicine on CardioCerebrovascularvol 15 pp 3151ndash3153 2017
[6] W J Yang and P Wang ldquoIntervention study of Huanglian jiedudecoction on obese patients with type 2 diabetes mellitusrdquoShandong Journal of Traditional Chinese Medicine vol 32 pp535ndash537 2013
[7] N Sekiya N Shibahara I Sakakibara N Hattori H Goto andK Terasawa ldquoInhibitory effects of oren-gedoku-to (Huanglian-Jie-Du-Tang) on free radical-induced lysis of human red bloodcellsrdquo Phytotherapy Research vol 17 pp 147ndash151 2003
[8] R X Fang Z X Liu and X H Zhong ldquoAnalysis of clinicaleffect serum cytokines VEGF and NO in unstable anginapectoris patients treated with Huang-Lian Jie-Du decoctioncombined with atorvastatinrdquo Pharmacology and Clinics of Chi-nese Materia Medica vol 36 pp 229ndash232 2017
[9] Q M Chu W C Wei Z Jin and W Wu ldquoPost-treatment ofhuanglian jiedu decoction on myocardial infarction patientswith acute ST segment elevationrdquo in Chinese Archives of Tra-ditional Chinese Medicine vol 36 pp 823ndash826 2018
[10] Y Xu M Qin Y K Zhao and Y Liu ldquoThe effect of cholesterollowing and anti-atherosclerosis of hanglian jiedu tang andits componentsrdquo Chinese Journal of Experimental TraditionalMedical Formulae vol 14 pp 74ndash77 2008
[11] J-M Brusq N Ancellin P Grondin et al ldquoInhibition of lipidsynthesis through activation of AMP kinase an additionalmechanism for the hypolipidemic effects of berberinerdquo Journalof Lipid Research vol 47 no 6 pp 1281ndash1288 2006
[12] O S Kim C-S Seo Y Kim H-K Shin and H Ha ldquoExtractsof scutellariae radix inhibit low-density lipoprotein oxidationand the lipopolysaccharide-inducedmacrophage inflammatory
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
SpectroscopyAnalytical ChemistryInternational Journal of
Hindawiwwwhindawicom Volume 2018
MaterialsJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
BioMed Research International Electrochemistry
International Journal of
Hindawiwwwhindawicom Volume 2018
Na
nom
ate
ria
ls
Hindawiwwwhindawicom Volume 2018
Journal ofNanomaterials
Submit your manuscripts atwwwhindawicom
International Journal of Analytical Chemistry 11
responserdquo Molecular Medicine Reports vol 12 no 1 pp 1335ndash1341 2015
[13] I-A Lee J H Lee N-I Baek and D-H Kim ldquoAntihyperlipi-demic effect of crocin isolated from the fructus of Gardeniajasminoides and its metabolite crocetinrdquo Biological amp Pharma-ceutical Bulletin vol 28 no 11 pp 2106ndash2110 2005
[14] Y Deng T Lu L Xie and X Liu ldquoHigh-performance liquidchromatographic method for the determination and pharma-cokinetic study of wogonoside in rat serum after oral admin-istration of traditional Chinese medicinal preparation Huang-Lian-Jie-Du decoctionrdquo Biomedical Chromatography vol 20no 10 pp 1098ndash1102 2006
[15] T Lu Y Liang J Song L Xie G J Wang and X DLiu ldquoSimultaneous determination of berberine and palmatinein rat plasma by HPLC-ESI-MS after oral administration oftraditional Chinese medicinal preparation Huang-Lian-Jie-Dudecoction and the pharmacokinetic application of the methodrdquoJournal of Pharmaceutical and Biomedical Analysis vol 40 no5 pp 1218ndash1224 2006
[16] H Zhu Z Qian H Li et al ldquoIntegrated pharmacokinetics ofmajor bioactive components in MCAO rats after oral adminis-tration of Huang-Lian-Jie-Du-Tangrdquo Journal of Ethnopharma-cology vol 141 no 1 pp 158ndash169 2012
[17] L B Yu Y Chen G L Xu et al ldquoStudy on mechanism ofhuang-lian jie-du decoction on atherosclerosis rats based onanti-inflammatory and antioxidantrdquo Modernization of Tradi-tional Chinese Medicine and Mateia Medica-World Science andTechnology vol 19 pp 1841ndash1845 2017
[18] L B YuG L Xu R Yao J LHu JNDuan andL Jiang ldquoEffectof huanglian jiedu decoction on atherosclerosis rats based onblood gas analysisrdquo Lishizhen Medicine and Materia MedicaResearch vol 28 pp 818ndash821 2017
[19] S Wu A Sun and R Liu ldquoSeparation and purification ofbaicalin and wogonoside from the Chinese medicinal plantScutellaria baicalensis Georgi by high-speed counter-currentchromatographyrdquo Journal of Chromatography A vol 1066 no1-2 pp 243ndash247 2005
[20] K Yu Y F Gong Z Y Lin and Y Y Cheng ldquoQuantitativeanalysis and chromatographic fingerprinting for the qualityevaluation of Scutellaria baicalensis Georgi using capillary elec-trophoresisrdquo Journal of Pharmaceutical andBiomedical Analysisvol 43 pp 540ndash548 2007
[21] J H Chen F M Wang J Liu S C Lee X R Wangand H H Yang ldquoAnalysis of alkaloids in Coptis chinensisFranch by accelerated solvent extraction combined with ultraperformance liquid chromatographic analysis with photodiodearray and tandem mass spectrometry detectionsrdquo AnalyticaChimica Acta vol 613 pp 184ndash195 2008
[22] S Pfister P Meyer A Steck and H Pfander ldquoIsolation andstructure elucidation of carotenoid-glycosyl esters in gardeniafruits (Gardenia jasminoides Ellis) and saffron (Crocus sativusLinne)rdquo Journal of Agricultural and Food Chemistry vol 44 no9 pp 2612ndash2615 1996
[23] J Sun J SMa J Jin et al ldquoQualitative and quantitative determi-nation of the main components of huanglianjiedu decoction byHPLC-UVMSrdquo Acta Pharmaceutica Sinica B vol 41 pp 380ndash384 2006
[24] S-K Ku and J-S Bae ldquoBaicalin baicalein and wogonin inhibitshigh glucose-induced vascular inflammation in vitro and invivordquo BMB Reports vol 48 no 9 pp 519ndash524 2015
[25] Y-Z Yang Y-Z Tang and Y-H Liu ldquoWogonoside dis-plays anti-inflammatory effects throughmodulating inflamma-tory mediator expression using RAW2647 cellsrdquo Journal ofEthnopharmacology vol 148 no 1 pp 271ndash276 2013
[26] S Lee Y Jeong M H Yu et al ldquoWogonin suppresses TNF-120572-induced MMP-9 expression by blocking the NF-120581B activationvia MAPK signaling pathways in human aortic smooth musclecellsrdquo Biochemical and Biophysical Research Communicationsvol 351 no 1 pp 118ndash125 2006
[27] Y Shi J Hu J Geng et al ldquoBerberine treatment reducesatherosclerosis by mediating gut microbiota in apoE-- micerdquoBiomedicine amp Pharmacotherapy vol 107 pp 1556ndash1563 2018
[28] N Ning K He Y Wang et al ldquoHypolipidemic effect andmechanism of palmatine from Coptis chinensis in hamsters fedhigh-fat dietrdquo Phytotherapy Research vol 29 pp 668ndash673 2015
[29] B Yan D Wang S Dong et al ldquoPalmatine inhibits TRIF-dependent NF-120581B pathway against inflammation inducedby LPS in goat endometrial epithelial cellsrdquo InternationalImmunopharmacology vol 45 pp 194ndash200 2017
[30] M Feng Study on the Role and Mechanisms of Alkaloids of Rhi-zoma Coptidis and Its Derivative on Atherosclerosis SouthwestUniversity 2017
[31] Y Yang H J Wang J Yang et al ldquoChemical profiling andquantification of Chinese medicinal formula Huang-Lian-Jie-Du decoction a systematic quality control strategy usingultra high performance liquid chromatography combined withhybrid quadrupole-orbitrap and triple quadrupole mass spec-trometersrdquo Journal of Chromatography A vol 1321 pp 88ndash992013
[32] K-Y Kwok J Xu H-M Ho et al ldquoQuality evaluation ofcommercial Huang-Lian-Jie-Du-Tang based on simultaneousdetermination of fourteen major chemical constituents usinghigh performance liquid chromatographyrdquo Journal of Pharma-ceutical and Biomedical Analysis vol 85 pp 239ndash244 2013
[33] Y X Deng T Lu L Xie and X D Liu ldquoHigh-performanceliquid chromatographic method for the determination andpharmacokinetic study of wogonoside in rat serum after oraladministration of traditional Chinese medicinal preparationHuang-Lian-Jie-DudecoctionrdquoBiomedChromatogr vol 20 pp1098ndash1102 2006
[34] M-Y He Y-X Deng Q-Z Shi X-J Zhang and Y LvldquoComparative pharmacokinetic investigation on baicalin andwogonoside in type 2 diabetic and normal rats after oraladministration of traditional Chinese medicine HuanglianJiedu decoctionrdquo Journal of Ethnopharmacology vol 155 no 1pp 334ndash342 2014
[35] M-F Zeng L-M Pan H-X Zhu Q-C Zhang and L-W GuoldquoComparative pharmacokinetics of baicalin in plasma after oraladministration of Huang-Lian-Jie-Du-Tang or pure baicalin inMCAO and sham-operated ratsrdquo Fitoterapia vol 81 no 6 pp490ndash496 2010
[36] H Zhu Z Qian F He et al ldquoNovel pharmacokinetic studies ofthe Chinese formula Huang-Lian-Jie-Du-Tang in MCAO ratsrdquoPhytomedicine vol 20 no 10 pp 767ndash774 2013
[37] The State Pharmacopoeia Commission of PR China ldquoMethod-ology validation guidelines for quantitative analysis of biologi-cal samplesrdquo in Pharmacopoeia of the Peoplersquos Republic of Chinavol IV pp 363ndash368 Chin Med Sci Press Beijing China 2015edition 2015
[38] X RGu S Y FangW Ren et al ldquoPharmacodynamics ofHuan-glian Jiedu decoction in Alzheimerrsquos disease (AD) model ratsand effect on improvement of inflammationmicroenvironment
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
SpectroscopyAnalytical ChemistryInternational Journal of
Hindawiwwwhindawicom Volume 2018
MaterialsJournal of
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
BioMed Research International Electrochemistry
International Journal of
Hindawiwwwhindawicom Volume 2018
Na
nom
ate
ria
ls
Hindawiwwwhindawicom Volume 2018
Journal ofNanomaterials
Submit your manuscripts atwwwhindawicom
12 International Journal of Analytical Chemistry
in brainrdquo China Journal of Chinese Materia Medica vol 43 pp3006ndash3011 2018
[39] F Zuo N Nakamura T Akao and M Hattori ldquoPharmacoki-netics of berberine and its main metabolites in conventionaland pseudo germ-free rats determined by liquid chromatogra-phyion trap mass spectrometryrdquo Drug Metabolism amp Disposi-tion vol 34 Article ID 011361 pp 2064ndash2072 2006
[40] S Yu X Pang Y Deng et al ldquoA sensitive and specific liquidchromatography mass spectrometry method for simultaneousdetermination of berberine palmatine coptisine epiberberineand jatrorrhizine from Coptidis Rhizoma in rat plasmardquo Inter-national Journal of Mass Spectrometry vol 268 no 1 pp 30ndash372007
[41] X Tan J Ma R Feng et al ldquoTissue distribution of berberineand its metabolites after oral administration in ratsrdquo PLoS ONEvol 8 no 10 p e77969 2013
[42] C Zhang R Yu Y Liu et al ldquoInteraction of baicalin withberberine for glucose uptake in 3T3-L1 adipocytes and HepG2hepatocytesrdquo Journal of Ethnopharmacology vol 151 no 2 pp864ndash872 2014
[43] L Jiang J D Dai Z L Huang Q H Du J H Lin andY R Wang ldquoSimultaneous determination of gastrodin andpuerarin in rat plasma by HPLC and the application to theirinteraction on pharmacokineticsrdquo Journal of ChromatographyB vol 915ndash916 pp 8ndash12 2013
[44] D A Chistiakov Y V Bobryshev E Kozarov I A Sobeninand A N Orekhov ldquoRole of gut microbiota in the modulationof atherosclerosis-associated immune responserdquo Frontiers inMicrobiology vol 6 p 671 2015
TribologyAdvances in
Hindawiwwwhindawicom Volume 2018
Hindawiwwwhindawicom Volume 2018
International Journal ofInternational Journal ofPhotoenergy
Hindawiwwwhindawicom Volume 2018
Journal of
Chemistry
Hindawiwwwhindawicom Volume 2018
Advances inPhysical Chemistry
Hindawiwwwhindawicom
Analytical Methods in Chemistry
Journal of
Volume 2018
Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018
