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REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 544
A REVIEW ON RECENT ADVANCEMENT OF LC-MS: ULTRA
HIGH PRESSURE LIQUID CHROMATOGRAPHY-MASS
SPECTROMETRY (UHPLC-MS) AND ITS APPLICATIONS
1Vishal Modi*, 2Akash Dubey, 3Parixit Prajapati, 1Tarashankar Basuri
1Head of Department, Department of Pharmaceutical Chemistry, SSR College of Pharmacy, sayli road, silvassa-396230, INDIA
2Department of Quality Assurance Techniques, SSR College of Pharmacy, sayli road, silvassa-396230, INDIA
3Department of Pharmaceutical Chemistry, SSR College of Pharmacy,
sayli road, silvassa-396230, INDIA
Corresponding Author:
Vishal Modi
Department of Pharmaceutical Chemistry,
SSR College of Pharmacy,
sayli road, silvassa-396230, INDIA
E-mail: vishalmodi1111@gmail.com
Phone: 9824931330
International Journal of Innovative
Pharmaceutical Sciences and Research www.ijipsr.com
Abstract
Nowadays, pharmaceutical manufacturer trying to reduce the cost and shorten the time of drug development. Focus is placed on recent developments in high efficiency LC separations, sensitive electro spray ionization approaches, and the benefits of amalgamating Liquid
chromatography-mass spectrometer (LC-MS) based approaches. The new high throughput Ultra high pressure liquid chromatography-mass spectrometer (UHPLC-MS) and/or
UHPLC/MS/MS focuses on shortening the overall analysis time, with higher resolution, high speed, high sensitivity, and increase separation efficiency .Metabolic profiling for screening analysis and detection and quantifying of very potent drugs mainly illicit drugs and drugs of
abuse in whole blood is made easier by this technique. With the availability of sensitive mass selective instruments, such as tandem mass spectrophotometers and high resolution unwanted
substance with minimal sample preparation.
Keywords: UHPLC-MS, LC-MS, Electro spray ionization.
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 545
INTRODUCTION
High-performance liquid chromatography (HPLC) is one of the main analytical techniques used
for drug analysis. As such HPLC meets many important criteria for analysis but it is time-
consuming and uses a high amount of solvents compared to other analytical techniques. Ultra-
high performance liquid chromatography (UHPLC) has been proposed as an alternative to HPLC.
The use of UHPLC systems for potent molecule analysis has gained widespread acceptance
among researchers. A UHPLC System is a HPLC system innovated with regards to dead volume,
detector sampling rate, injection Performance and is able to tolerate application Pressures up to
1,500 bar. It is been improved in areas like chromatographic resolution, sensitivity of analysis etc.
It is therefore advantageous to use UHPLC instrumentation for increasing demand to provide
faster and selective screening techniques for an expanding range of drugs for forensic
methodology and workplace drug screening. Reduced analysis time provide for rapid release of
data, reduced costs per assay, and greater sample throughput overall.
Other important methodology exploit the high mass resolution capabilities of specially designed
mass spectrometers where these are not easily available, and where precursor and product ions
both are present ,so qualification and quantification of potent analytes is made easier by UHPLC-
MS [1].
PRINCIPLE
The basic principle of this chromatographic technique is governed by an formula known as Van
Dee meter equation which shows the relationship between plate height (HETP or column
efficiency) and linear velocity (flow rate). According to this equation, the increase in efficiency of
UHPLC technique is not possible without reducing particle size than those used in conventional
HPLC technique.
The Van Deemeter equation:
H = A + B/v + Cv,
Where, and A, B and C are constants and v is linear velocity.
Here, A represents the Eddy mixing which is independent of velocity. The value of A is lowest
when column particles are uniformly small.
B is the natural diffusion tendency of molecules or axial diffusion which is affected by flow rate.
This effect is diminished at high flow rates, so this term is divided by v.
C is the kinetic resistance to equilibrium in the separation process [2-6].
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
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Fig. 1: diagram showing importance of HETP in column efficiency
INSTRUMENTATION
There has been great interest in approaches to rapid up and/or increase the resolving power of the
analytical separation process, particularly with the making of columns packed with porous sub-
2μm particles which is to be used in very high pressure conditions. Use of small particle size
results in higher plate numbers, as well as rapid separations.
These effects are due to;
i) The chromatographic efficiency, N, is directly proportional to the particle diameterand
ratio of column length, L/dp..
ii) The mobile phase linear velocity, u, is inversely proportional to the (dp) particle diameter.
Generally, the particle size reduction generates a high backpressure (more than 400 bar) which is
very difficult with conventional instrumentation. Therefore, to benefit from the full potential of
columns packed with small particles, it is smarter to work with a chromatographic system that
withstands pressures up to 1000 bar [7].
REQUIREMENTS FOR UHPLC EXPERIMENTS
PUMPING SYSTEM
For achieving high peak capacity separations requires a higher pressure range than that obtainable
in HPLC instrumentation. Across a 15 cm long column packed with 1.7 um particles the
calculated pressure drop at the optimum flow rate for maximum efficiency is about 15,000 psi.
Therefore a pump must be capable of delivering solvent reproducibly and smoothly at these
pressures, which can operate in both the gradient and isocratic separation modes. Up to 15,000-psi
pressure limit (about 1000 bar) can be achieved to take full advantage of the sub 2 um particles
[8,9].
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
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SAMPLE INJECTION
Sample introduction in UHPLC is very important. Injection valves which are used conventionally
either manual or automated are not designed to work at extreme pressure. Protection of the
column from extreme pressure fluctuations, the process of injection must be pulse-free and the
swept volume of the device required being very low to reduce band spreading. Rapid injection
cycle time is required to fully capitalize on the speed afforded by UHPLC, which in turn need a
high sample capacity. Least carryover with low volume injections are also needed to increase
sensitivity. Direct injection approaches are also there for biological samples.10-11
SAMPLE MANAGER
With the help of high pressure of sample introduction low dispersion is maintained through the
injection process and diagnostics as well as self-monitoring can be facilitated by pressures
transducers in series. Variety of micro titer plate formats (mid height, deep well or vials) can also
be used in a thermostatically controlled environment. Using the particular sample organizer, the
sample manager can inject up to 22 micro titer plates. The column heater is also controlled by
sample manager. Column temperatures in a range up to 65°C can be maintained. A “pivot out”
design allows the column outlet to be placed in nearer proximity to the source inlet of an MS
detector to reduce sample dispersion.12
QUALITY OF MOBILE PHASE AND BUFFERS
Ultra high purity solvents signify directly to the sensitivity of the mass spectrometer that
reduces chromatographic interferences. It is important to verify the absence of insoluble
particles in the solvents and, for this purpose; many important rules for the mobile phase
preparation have to be followed in UHPLC.
Use only high grade organic solvents: methanol or acetonitrile (ideally filtered through a
0.22 μm membrane) – it is possible to obtain acetonitrile of UHPLC grade from several
suppliers (BIOSOLVE®, FISHER SCIENTIFIC®, JT BAKER® etc…)
Use of high quality salts for preparation of buffered mobile phases
The water must be ultra-pure and filtered through a 0.22 μm membrane (Milli-Q® system
of similar high quality water is recommended)
Benefit: Submicron filtration ensures minimal clogging of instrument, columns and check valves
Minimizes the leaching of metal cations (Na+ and K+).
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 548
Benefit: Low metal content in mobile phase solvents reduces formation of metal ion adducts. Be
careful from the microbiological growth (mainly when using phosphate buffer): Use only freshly
prepared mobile phases. Be vigilant with the cleaning of glassware and do not top-off the bottles.
The chromatographic system and columns should ideally be stored with pure organic solvents
(methanol or acetonitrile) to limit the microbiological growth [7].
UHPLC COLUMNS
Resolution can be increased in small 1.7 μm particle packed column because efficiency is better
with use of smaller particles. Sample requires a bonded phase for the separation of the
components that provides both retention and selectivity. Column length should be selected
according to the needed efficiency (in isocratic mode) or peak capacity (in gradient mode) [7].
List of different columns used in UHPLC
ACE EXCEL 2 C18
ZORBAX ECLIPSE 1.8 XDB C18
WATERS ACQUITY 1.7 BEH C18
PHENOMENEX KINETEX 1.7 C18
ACE EXCEL 2 C18-ARACE EXCEL 2 C18-PFP
High Efficiency UHPLC columns are available in 2μm particle size. Optimized UHPLC columns
utilize incredibly developed low dispersion hardware which enables high efficiency UHPLC
separations up to a maximum pressure limit up to 1,000 bar (15,000psi). UHPLC columns are
also available with C4, CN, AQ, C18, C8, Phenyl and Silica phases. C18 bonded phases are the
most popular phases for UHPLC because they provide high selectivity and good retention for a
wide variety of sample types. In addition, they are more rugged and reliable than other bonded
phases. C18 bonded phases mainly depends on hydrophobic interactions (and sometimes shape
selectivity) to achieve satisfactory separations. But, C18 phases offer a limited number of
separation mechanisms which may lead to less than optimum separations in some cases and
complete lack of resolution of important peak pairs. ACE columns have offered C18 bonded
phases with the advantage of „extra selectivity‟. C18-PFP and C18-AR bonded phases have
proved to be extremely powerful tools to use in leveraging this „extra selectivity‟ to obtain
separations that may not be possible with „standard‟ C18 bonded phases. The introduction of
different UHPLC columns means that chromatographers now have more options within their
reach to achieve great results with their UPLC and UHPLC instruments [13,14].
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 549
Fig. 2: diagram of column used in UHPLC
Advantages of UHPLC
• Increases sensitivity.
• Selectivity, sensitivity, and dynamic range of LC analysis drastically increased..
• Resolution performance is maintained.
• Enhance scope of Multi residue Methods.
• UHPLC‟s rapid resolving power quickly quantifies related and unrelated compounds.
• Quick analysis by using of a new separation material of very fine particle size.
• Operation cost is can be reduced.
• Solvent consumption in reduced.
• Real-time analysis in step with manufacturing processes can be delivered.
• Assures end-product quality and final release testing [5,6].
Disadvantages of UHPLC
Requires more maintenance and reduces the life of the columns due to increased pressure column
of this type. So far performance similar or even higher has been reported by using stationary
phases of size up to 2 μm without the adverse effects of high pressure [5,6].
MASS SPECTROMETRY INSTRUMENTATION
Operation of mass spectrometer is based on converting the analyte molecules to a charged
(ionized) state, with frequently analysis of the ions and any fragment ions that are obtained during
the ionization process, on the basis of their mass to charge ratio (m/z). Many different
technologies are available for both ionization and ion analysis, resulting in many different types
of mass spectrometers with different combinations of these two processes.
Fig. 3: various components of mass spectrometer
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 550
ION SOURCES
ELECTROSPRAY IONIZATION SOURCE
ESI was developed by Fenn into a robust ion source which is capable of interfacing with highly
optimized LC and demonstrated its application to a number of important classes of biological
molecules. ESI works significant with moderately polar moieties and is thus well suited to the
analysis of many metabolites, drugs and peptides. Pumping of a liquid sample through a metal
capillary maintained at 3 to 5 kv and nebulized at the tip of the capillary to form a charged
droplets which are very fine.
The capillary is usually off-axis from, or orthogonal to, the entrance to the mass spectrometer in
order to reduce contamination. The droplets are quickly evaporated by the help of dry nitrogen
and heat, and the transfer of residual charge to the analytes from the charged droplets takes place
[15].
Later the ionized analytes are transferred from the high vacuum to the mass spectrometer by
series of tiny apertures and focusing voltages. The subsequent ion optics and ion source can be
functioned to detect positive or negative ions, and switching between these two modes within an
analytical run can be performed. Under general conditions, ESI is known as “soft” ionization
source, meaning that relatively low energy is imparted to the analyte, and hence little
fragmentation occurs.
This has been used in LC-MS analysis to identify components with common structural features
e.g. Glycopeptides from glycan‟s can be fragmented in-source to give 204 m/z reporter ions. This
feature has been used to identify glycopeptides in tryptic digests of proteins in order to
characterize the structure of the glycan‟s.
While ESI is the most widely used ion source for biological molecules, neutral and low polarity
molecules like lipids may not be efficiently ionized by this method.
ATMOSPHERIC PRESSURE CHEMICAL IONIZATION SOURCE
As with ESI, in atmospheric pressure chemical ionization (APCI), liquid is pumped from a
capillary and nebulized at the tip. A corona discharge taking place near the tip of the capillary,
initially solvent molecules and ionizing gas present in the ion source.
Ions then react with the analyte and ionize it via charge transfer. The technique is useful for
thermally stable and small molecules which are not well ionized by ESI. The technique has also
been applied to fat soluble vitamins and lipids [16,17].
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 551
Fig. 4: schematic diagram of atmospheric pressure chemical ionization source
MASS ANALYSERS
QUADRUPOLE ANALYSERS
A set of four parallel metal rods are present in quadrupole analyser. A combination of varying
(radio frequency) and constant voltages allows the transmission of a narrow band of m/z values
along the axis of the rods. It is possible to scan across a range of m/z values, resulting in a mass
spectrum by varying the voltages with time. Most quadrupole analysers scan speeds up to 1000
m/z per sec or more are common and operate at <4000 m/z. They usually operate at unit mass
resolution meaning that the mass accuracy is better than 0.1 m/z. Alternative to scanning, the
quadrupole can be set to monitor with a specific m/z value, then set to monitor another m/z value,
and soon. This is achieved by stepping the voltages. This technique is important for improving the
detection limits of specific targeted analytes because more detector time can be reduced for
detecting specific ions in place of scanning across ions that are not produced by the analyte.
Stepping can be carried out within few milliseconds and a panel of m/z values can be stepped for
the detection of many analytes. Ions can be induced to undergo fragmentation by collisions with
an inert gas such as argon or nitrogen, a process known as collision induced dissociation.
Quadrupole is one type of collision cell that has been designed to maintain the low pressure of the
collision gas needed for dissociation and transmit most of the fragment ions which are produced.
Mass spectrometer configurations which are particularly useful are obtained by placing a collision
cell between two quadrupole mass analysers. This combination is known as triple quadrupole
mass spectrometer which is an example of tandem MS in which independently application of two
or more stages of mass analysis are achieved. The advantage of tandem MS is greatly increased
specifically over the analysis carried out by single stage mass analysis. For example, 25-hydroxy
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 552
vitamin D3 gives 401 m/z as major M+H+ ion during ESI that loses water at the time of collision
induced dissociation to give a major 383 m/z product ion.12 Highly specific detection of 25
hydroxy vitamin D3 is possible if the first and third quadrupoles are set to transmit ions of only
401 and 383 m/z respectively. This method is termed as single reaction monitoring and the
fragmentation is denoted 401>383 [19].
Fig. 5: schematic diagram of quadrapole mass analyser
TIME-OF-FLIGHT ANALYSERS
The Time-of-Flight (TOF) analyser works by accelerating ions through a high voltage. The time
needed to travel down a flight tube to reach the detector and velocity of ions, depends on their m/z
values. The output of the detector as in term of time can be converted into a mass spectrum if the
initial accelerating voltage is pulsed.
The TOF analyser can gain spectra extremely rapid with high sensitivity. It also has high mass
accuracy, through which molecular formulas of small molecules can be determined [19].
ION TRAP ANALYSERS
Ion trap analysers apply three hyperbolic electrodes to trap ions in a 3-D space using radio
frequency and static voltages. Ejection of ions sequentially from the trap on behalf of their m/z
values to create a mass spectrum takes place.
Alternatively, by the application of an exciting voltage, a specific ion can be isolated in the trap
while other ions are ejected. An inert gas can also be entered into the trap to induce
fragmentation. An important feature of these ion trap analysers is the ability to isolate and
fragment ions many times in succession before the final mass spectrum is obtained, resulting in
so-called msn capabilities [19,20].
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
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VARIOUS APPLICATIONS OF UHPLC-MS:
TESTING AND QUANTIFICATION OF ILLICIT DRUGS IN URINE USING UHPLC
COUPLED TO ACCURATE MASS AXION 2 TOF MASS SPECTROMETER.
LC CONDITIONS:
LC System:
Pump: PerkinElmer Flexar™ FX-10 pump
Flow: 0.4 ml/min
Mobile phase A: Water containing with 0.1% formic acid
Mobile phase B: Acetonitrile containing with 0.1% formic acid
Injection volume: 4 μl in partial fill mode
Column used: PerkinElmer Brownlee™ SPP C-18, 2x50 mm, 2.7 μm at 25 °C
MS Conditions:
Mass spectrometer: PerkinElmer axion 2 TOF MS
Ionization source: PerkinElmer Ultra spray™ 2 (Dual ESI source)
Ionization mode: Positive
Spectral acquisition rate: 3 spectra/sec
Capillary exit voltage: 100 V
Trap pulse™ mode: 100-1000 m/z
Table 1: Testing and quantification of illicit drugs in urine
Sl.
No
.
Analyte Formula Monoisotopic
Mass [M+H]+
Observed
Mass
Mass
Error
(ppm) 1 Methamphetamine C10H15N 150.277 150.1273 -2.66
2 Amiodarone C25H29I2NO3 646.0310 646.0312 0.39 3 Amphetamine C19H13N 136.1121 1361113 -5.88
4 Codeine C18H21NO3 300.1594 300.1594 -0.07 5 Diazepam C16H13Cln2o 285.0789 285.0792 0.98
6 Doxepine C19H21NO 280.1696 280.1695 -0.36 7 Haloperidol C21H23clfno2 376.1474 376.1474 0.00
8 Morphine C17H19NO3 286.1438 286.1434 -1.40 9 Flurazepam C21H23clfn3o 388.1586 388.1593 1.80
10 Alprolazem C17H13cln4 309.092 309.0900 -0.65 11 Cocaine C17H21NO4 304.1543 304.1550 2.30
12 EDDP perchlorate
(fragment, [M-O4Cl]+) C20H24Clno4 278.1903 278.1906 1.08
13 Benzoyl ecognine C16H19NO4 290.1387 290.1394 2.41
14 Methadone C21H27NO 310.2165 310.2171 1.93 15 3,4 MDA C10H13NO2 180.1019 180.1016 -1.67
16 MDEA C12H17NO2 208.1332 208.1326 -2.88
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
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17 MDMA C11H15NO2 194.1176 194.1169 -3.61 18 Phenyl propyl amine C9H13NO 152.1069 152.1060 -5.92
19 Oxycodone C18H21NO4 316.1543 316.1545 0.54
UHPLC-ESI-MS METHOD FOR THE QUANTIFICATION OF STEROID HORMONES.
EXPERIMENTAL PREPARATION OF STANDARD SOLUTION:
Steroid solution (single standard) of 1 mg/ml was prepared in 50:50 (v/v) water acetonitrile.
Method Parameters
Column: blue orchid 175-1.8 C18, 50 x 2 mm ID
Eluent A: Water+ 0.1 % Formic Acid
Eluent B:Acetonitrile + 0.1 % Formic Acid
Flow rate 0.4 ml/min
Injection volume 5 μl
Column temperature 40 °C
Run time3.0 min
MS Conditions:
Mass spectrometer : MSQ Plus single quadrapole mass spectrometer
Ionization mode : positive mode, ESI
Needle Voltage : 3.5 kv
Cone Voltage : 20 V
Probe temperature : 350 °C
MS Detection parameters
Mode Full scan mode, m/z 270 – 370
RESULT
For MS detection, the mass spectra for every compound obtained. The highest intensity fragment
is chosen for quantification. Fragment ions having lower intensities can be used as qualifiers in
real samples.
Table 2: Quantification of steroid hormones
Steroid Ionization Mass
[g/mol]
M/z Expected
[M+H]+
M/z
Found
Corticosterone ESI + 346.4 347 347
Cortisone ESI + 360.4 361 361
Deoxycorticosterone ESI + 330.5 331 331
Norgestrel ESI + 312.3 313 313
Progesterone ESI + 314.5 315 315
Testosterone ESI + 288.4 289 289
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
Department of Quality Assurance ISSN (online) 2347-2154
Available online : www.ijipsr.com May Issue 555
IDENTIFICATION OF CANNABINOIDS IN BAKED GOODS BY UHPLC/MS.
EXPERIMENTAL CONDITIONS:
Standard and Sample Preparation were mixed and diluted to up to 10 ppm to prepare a stock
solution with methanol.
Chromatographic Conditions:
Chromatographic analyses were performed using the Accela UHPLC system (Thermo Fisher
Scientific)
Column: Hypersil GOLD PFP (per fluorinated phenyl) 1.9 μm, 100 x 2.1 mm
Flow Rate: 1 ml/min
Mobile Phase: A: Water with 0.06 % acetic acid
B: Acetonitrile with 0.06% acetic acid
C: Methanol with 0.06% acetic acid
Column Temperature: 45 °C
Injection:
Syringe Speed: 8 μl/sec
Flush Speed: 100 μl/sec
Flush Volume: 400 μl
Wash Volume: 100 μl
Flush/Wash Source: Bottle with methanol
Mass Spectrometer Conditions
MS analysis was carried out on MSQ Plus single quadrapole LC/MS detector with Xcalibur 2.05
(Thermo Fisher Scientific).
The MS conditions were as given:
Ionization: ESI
Polarity: Positive
Probe Temperature: 500 °C
Cone Voltage: 90 V
Scan Mode: mass range of 50-500 m/z with full scan
ESI Voltage: 3.5 kv
Scan Time: 0.2 s H
The cannabinoids were detected by using full scans (50-500 m/z) of the single quadrupole mass
spectrometer, and the extracted ion chromatograms from m/z310.5-311.5 + 314.5-315.5
REVIEW ARTICLE Vishal et.al / IJIPSR / 4 (5), 2016, 544-558
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Table 3: Identification of cannabinoids in baked goods
COMPOUND
RT min
(STANDARD
COMPOUND)
RT
(BROWNIE)
Min
RT
(COOKIE)
Min
Product
ion (m/z)
Product
ion (m/z)
Brownie
Product
ion (m/z)
Cookie
Cannabidol 4.1 -- -- 315.31 -- --
THC 5.1 5.1 5.1 315.33 315.33 315.27 Cannabinol 5.4 -- -- 311.31 -- --
Using UHPLC/MS Cannabinoids in baked goods can be identified with minimal sample
preparation. The preparation time up to 10 min and run time 8 min make this a very efficient
analytical method.
QUALITATIVE AND QUANTITATIVE ANALYSIS OF UHPLC-MS
Determination of polyphenols
Determination of pesticides
Determination of primary aromatic amines
Determination of benzodiazepines
Determinations of aflatoxins
Determination of coumarin and chlorogenic acid.
Mycotoxin analysis.
CONCLUSION
UHPLC-ms give increased resolution, speed and sensitivity for liquid Chromatography-mass
spectrometry. The main advantage of UHPLC-ms is a reduction of analysis time, along with
reduced solvent consumption, high throughput analysis and reduction in cost of analysis. From
the literature survey it can be concluded that all categories of pharmaceutical drugs can be
analyzed by UHPLC-ms method within a very short period of time and with less solvent
consumption. UHPLC-ms technology is transforming lives and laboratories, creating new
opportunities for business profitability, and bringing new meaning to quality. The literature
survey shows that research on UHPLC-ms analysis, both, at national and international level have
been successfully done on all categories of drugs.
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