5-----GCMS analysis of the rat urine for metabonomic research

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1、JournalofChromatographyB,854(2007)2025GC/MSanalysisoftheraturineformetabonomicresearchQiZhanga,b,GuangjiWanga,∗,YuDub,LinglingZhub,JiyeAc,∗∗aCenterofDrugMetabolismandPharmacokinetics,ChinaPharmaceuticalUniversity,Nanjing,ChinabCollegeofLifeandPharmacy,NanjingUniversityofTechnology,Nanjin

2、g,ChinacDepartmentofMedicalBiosciences,ClinicalChemistry,Ume˚aUniversity,SE-90185Ume˚a,SwedenReceived24September2006;accepted26March2007Availableonline8April2007AbstractInthispaper,anoptimizedprotocolwasestablishedandvalidatedforthemetabonomicprofilinginraturineusingGC/MS.Theurinesam-ples

3、wereextractedbymethanolaftertreatmentwithureasetoremoveexcessiveurea,thentheresultedsupernatantwasdried,methoximated,trimethylsilylated,andanalyzedbyGC/MS.Forty-nineendogenousmetaboliteswereseparatedandidentifiedinGC/MSchromatogram,ofwhich26identifiedcompoundswereselectedforquantitativeana

4、lysistoevaluatethelinearity,precision,andsensitivityofthemethod.Itshowedgoodlinearitybetweenmassspectrometryresponsesandrelativeconcentrationsofthe26endogenouscompoundsovertherangefrom0.063to1.000(v/v,urine/urine+water)andsatisfactoryreproducibilitywithintra-dayandinter-daysprecisionvalu

5、esallbelow15%.ThemetabonomicprofilingmethodbasedonGC/MSwassuccessfullyappliedtourinesamplesfromhyperlipidemiamodelrats.Obviously,separatedclusteringofmodelratsandthecontrolratswereshownbyprincipalcomponentsanalysis(PCA);time-dependentmetabonomicmodificationwasdetectedaswell.Itwassuggestedt

6、hatmetabonomicprofilingbasedonGC/MSbearobustmethodforurinesamples.©2007ElsevierB.V.Allrightsreserved.Keywords:Metabonomics;Urine;GC/MS1.Introductionisthoughttheidealtoolformetabonomicprofilingsincemostmetabolitesinurineareofpolargroups,whichwillfacilitateMetabonomicsisasystematicapproachto

7、studyinvivoderivatizationandanalysisinGC/MS.metabonomicprofile,whichisdefinedasthequantitativemea-GC/MShasbeenappliedtometabonomicprofilingofurinesurementofthetime-relatedmultiparametricmetabonomicsamplesfromhydrocortisone-inducedanimalmodel[10],whereresponseoflivingsystemst