Gcell

Beijing Strong Biotechnologies, Inc.Add:No. 15, North Second Street, Yanqi Economic Development Area, Huairou District, Beijing,101400,P.R.ChinaTel: +86 10 6166 7168 Fax: +86 10 6166 7810

Web: www.bsbe.com.cn Email: tech@bsbe.com.cn Revised 01FEB11 version 1102

1,5-Anhydroglucitol1,5-Anhydroglucitol1,5-Anhydroglucitol1,5-Anhydroglucitol AssayAssayAssayAssay KitKitKitKit((((1,5-AG1,5-AG1,5-AG1,5-AG))))

Method:Method:Method:Method: IFCC Method Without pyridoxal-phosphateCaCaCaCatttt .... No.No.No.No. SSSSizeizeizeize InstrumentInstrumentInstrumentInstrument

GB8122T R1:3×20 mlR2:1×20 mlFor Hitachi 717&ShimadzuCL7200/8000

GS8123T R1:3×20 mlR2:1×20 ml

For Hitachi917&OlympusAU640/400/60

0

INTENDEDINTENDEDINTENDEDINTENDED USEUSEUSEUSEThe 1,5-Anhydroglucitol (1,5-AG) assay is usedfor the quantitation of 1,5-Anhydroglucitol inhuman serum or plasma.

CLINICALCLINICALCLINICALCLINICAL SIGNIFICANCESIGNIFICANCESIGNIFICANCESIGNIFICANCE1,5-Anhydroglucitol (AG; popularly called 1-deoxyglucose) has a pyranoid structure,resulting from the deletion of an oxygen fromglucose at the anomeric hydroxyl group. Thiscompound is one of the main sugar alcohols inhuman cerebrospinal fluid and serum. In plasma,the concentration of AG is reduced specificallyin diabetes mellitus, thus ninking it useful as adiagnostic marker for the disease.Because 1,5-AG is similar structures withglucose, due to high blood glucose brings theglucose excrete (diabetes), competition withglucose 1,5 - AG, make urine reuptake 1,5-AGemissions increase of serum concentrations,reduced. At this time, the body 1,5-AG storagepool also reduces, reduce blood glucose afterimprovement, with normally accepted from foodsupply returned to normal. 1,5-AG can sensitivereaction, blood sugar control as a highlysensitive state-of-the-art of glucose in bloodsugar, comprehensive index forecast changes,is determination can also mean days interval ofmeaningful change. Especially to grasp milddiabetics glucose change is very effective.ASSAYASSAYASSAYASSAY PRINCIPLESPRINCIPLESPRINCIPLESPRINCIPLESPyranonse oxidase, Hexokinase and ATPregenerative system that can detect the serum1,5 -AG concentration. The assay utilizesHexokinase and ATP regenerative systemconverts glucose to Glucose-6-phosphate, acompound that does not react with Pyranoseoxidase. The hydrogen peroxide produced inthe oxidation of 1,5-AG by Pyranose oxidase isdetected with a standard enzymatic color-developing system.

HexokinaseHexokinaseHexokinaseHexokinaseGGGGlucoslucoslucoslucos eeee ++++ ATPATPATPATP Glucose-6-phosphateGlucose-6-phosphateGlucose-6-phosphateGlucose-6-phosphate ++++ ADPADPADPADP

PyruvastePyruvastePyruvastePyruvaste KinaseKinaseKinaseKinasePEPPEPPEPPEP ++++ ADPADPADPADP PyruvatePyruvatePyruvatePyruvate ++++ ATPATPATPATP

PyranosePyranosePyranosePyranose OxidaseOxidaseOxidaseOxidase1,5-Anhydroglucitol1,5-Anhydroglucitol1,5-Anhydroglucitol1,5-Anhydroglucitol ++++ OOOO2222 1,5-anhydro-fructose1,5-anhydro-fructose1,5-anhydro-fructose1,5-anhydro-fructose ++++ HHHH2222OOOO2222

PODPODPODPODHHHH2222OOOO2222++++ 4-aminoantipyrine/HTIB4-aminoantipyrine/HTIB4-aminoantipyrine/HTIB4-aminoantipyrine/HTIB HHHH2222OOOO +Dye+Dye+Dye+Dye

SAMPLESAMPLESAMPLESAMPLE COLLECTIONCOLLECTIONCOLLECTIONCOLLECTION ANDANDANDAND PREPARATIONPREPARATIONPREPARATIONPREPARATIONSerum or plasma samples.Use fresh patient serum or plasma samples.

REAGENTREAGENTREAGENTREAGENT COMPOSITCOMPOSITCOMPOSITCOMPOSITIIIIONONONON

ContentsContentsContentsContents ConcentrationConcentrationConcentrationConcentration ofofofofSolutionsSolutionsSolutionsSolutionsReagentReagentReagentReagent 1111 (R1)(R1)(R1)(R1)

Mes Buffer(pH=6.3) 50mmol/LHexokinase 4KU/LATP 1mmol/LPEP 4mmol/LPyruvaste Kinase 3KU/L4-aminoantipyrine 1.5mmol/LAscorbrate oxidase 5KU/Lpreservative 0.1%

ReagentReagentReagentReagent 2222 (R2)(R2)(R2)(R2)Hepes Buffer (pH=8.0) 200mmol/LPyranose Oxidase 80KU/LPeroxidase 10KU/LHTIB 4.5mmol/Lpreservative 0.1%CalibrationControl

STABILITYSTABILITYSTABILITYSTABILITY ANDANDANDAND PREPARATIONPREPARATIONPREPARATIONPREPARATION OFOFOFOFREAGENTSREAGENTSREAGENTSREAGENTSAll reagents are ready to use.Stable up to the expiry date when stored at 2-8℃.Once opened the reagent is stable for 1 monthOn-board the analyser at approximately 10°C.

ASSAYASSAYASSAYASSAY PROCEDUREPROCEDUREPROCEDUREPROCEDURETestTestTestTest ProcedureProcedureProcedureProcedure forforforfor AnalyzersAnalyzersAnalyzersAnalyzers (HITACHI 917)Assay Mode: Rate A 16-31Wave Length (main/sub): 546 nm/ 700 nm

Sample: 6 μlR1: 180 μl R2: 60 μl

Measure Measure

37℃0 5 9 (min)

1. Mix 6μl sample with 180μl R1 and incubateat 37℃ for 5 minutes.

http://www.bsbe.com.cn/mailto:tech@bsbe.com.cn

Gcell

Beijing Strong Biotechnologies, Inc.Add:No. 15, North Second Street, Yanqi Economic Development Area, Huairou District, Beijing,101400,P.R.ChinaTel: +86 10 6166 7168 Fax: +86 10 6166 7810

Web: www.bsbe.com.cn Email: tech@bsbe.com.cn Revised 01FEB11 version 1102

2. Read initial absorbance A1.3. Add 60μl R2 into cuvette, mix and incubate

at 37℃ for 5 minute.4. Read initial absorbance A2.5. Calculate absorbance change (∆A=A2-A1).

CALCULATIONCALCULATIONCALCULATIONCALCULATION∆A sample

Concentration= × calibrator value∆Acalibrator

CALIBRATIONCALIBRATIONCALIBRATIONCALIBRATIONRecommend that this assay should becalibrated using Gcell Calibration.

QUALITYQUALITYQUALITYQUALITY CONTROLCONTROLCONTROLCONTROLGcell quality control, Level 1 and Level 2 arerecommended for daily quality control. Twolevels of controls should be assayed at leastonce a day. Values obtained should fall within aspecified range. If these values fall outside therange and repetition excludes error, thefollowing steps should be taken:1. Check instrument settings and light source.2. Check reaction temperature.3. Check expiration date of kit and contents.

NORMALNORMALNORMALNORMAL VALUEVALUEVALUEVALUE [1][1][1][1]Serum or plasma: ＞14 μg/mL（85.26 μmol/L）It is recommended that each laboratoryestablish its own reference range to reflect theage, sex, diet and geographical location of thepopulation.

LINEARITYLINEARITYLINEARITYLINEARITYThe method is linear between AGconcentrations of 10- 300 μmol/L. If the Sampleabove this concentration should be diluted itwith 0.9% NaCl and repeat assay.

SPECIFICSPECIFICSPECIFICSPECIFIC PERFORMANCEPERFORMANCEPERFORMANCEPERFORMANCECHARACTERISTICSCHARACTERISTICSCHARACTERISTICSCHARACTERISTICSINTERFERENCEINTERFERENCEINTERFERENCEINTERFERENCEThe following analytes were tested up to thelevels indicated and found not to interfere:Hb 200 mg/dLTG 500 mg/dLAscorbic Acid: 50 mg/dLGlucose 20 mmol/L

SENSITIVITYSENSITIVITYSENSITIVITYSENSITIVITYThe minimum detectable concentration of AGwith an acceptable level of precision wasdetermined as 7.8 umol/L.With serum sample 150umol/L test, as its everyminute changes spectrophotometry between in0.005 ~ 0.030.

PRECISIONPRECISIONPRECISIONPRECISIONThe CV of the test should be CV

Gcell

Beijing Strong Biotechnologies, Inc.Add:No. 15, North Second Street, Yanqi Economic Development Area, Huairou District, Beijing,101400,P.R.ChinaTel: +86 10 6166 7168 Fax: +86 10 6166 7810

Web: www.bsbe.com.cn Email: tech@bsbe.com.cn Revised 01FEB11 version 1102

http://www.bsbe.com.cn/mailto:tech@bsbe.com.cn

Gcell

Beijing Strong Biotechnologies, Inc.Add:No. 15, North Second Street, Yanqi Economic Development Area, Huairou District, Beijing,101400,P.R.ChinaTel: +86 10 6166 7168 Fax: +86 10 6166 7810

Web: www.bsbe.com.cn Email: tech@bsbe.com.cn Revised 01FEB11 version 1102

INSTRUMENTINSTRUMENTINSTRUMENTINSTRUMENT SETTINGSSETTINGSSETTINGSSETTINGS FORFORFORFOR HITACHIHITACHIHITACHIHITACHI 917917917917 INSTRUMENTINSTRUMENTINSTRUMENTINSTRUMENT SETTINGSSETTINGSSETTINGSSETTINGS FORFORFORFOR HITACHIHITACHIHITACHIHITACHI 902902902902

INSTRUMENTINSTRUMENTINSTRUMENTINSTRUMENT SETTINGSSETTINGSSETTINGSSETTINGS FORFORFORFOR OLYMPUSOLYMPUSOLYMPUSOLYMPUS 400/640/2700400/640/2700400/640/2700400/640/2700 INSTRUMENTINSTRUMENTINSTRUMENTINSTRUMENT SETTINGSSETTINGSSETTINGSSETTINGS FORFORFORFOR BECKMANBECKMANBECKMANBECKMAN CX4/5/7/9CX4/5/7/9CX4/5/7/9CX4/5/7/9

http://www.bsbe.com.cn/mailto:tech@bsbe.com.cn