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1、StudyoftheApplicationoftheGC-MSquantitativemethodindiagnosisofInbornErrorofMetabolismMDCandidateMinYanJiangSupervisorLiLiuAbstractObjective1.Toestablishaquantitativemethodformeasurementofurinaryabnormalmetabolitesbygaschromatography-massspectrometry(GC-MS);2.Toexamineid
2、entificationandquantitationofurinarymetabolitesbydifferentpre-treatmentmethods.Methods1.Toquantify12urinaryorganicacids,whichiskeydiagnosticmetabolitesbyGC-MS2.Toestablishcalibrationcurves.Quantificationisbycomparisontocalibrationofpurestandardcompoundsinratiotoanintern
3、alstandard・3.Validationdata,includingdetectionlimit,accuracy,precision,recoveryhavebeenstudiedandlinearityhasbeenevaluatedfor12organicacids.4.Measurementof12urinaryorganicacidsinnormalchildren.5.49positiveurinespecimensincludingorganicaciduria，aminoacidmetabolismdisorde
4、r,NICCDpatientand9positivecontrolsfromERNDIMwerepreparedpriortoGC/MSanalysis.6.Toanalyseallurinebyorganicsolventextractionandurease-pretreatmentpriortoGC-MS.7.ToidentifyeachcompoundintheurinebycomparingwiththestandardspectrumintheNISTlibrary.1.Toquantify12urinaryorganic
5、acidsbydifferentpre-treatmentmethodandtoanalysedetectionlimit,accuracy,precision,recovery,etc.2.Tocompareureasepretreatmentandorganicsolventextractionforanalyzingindentifiedmetabolitesqualitativelyandquantitativelybyt-test,ifP<0.05,thereisstatisticalsignificance.Results
6、1.Afterureasepre-treatment,derivatization,theretentiontimeof3・hydroxypropionicacid,methylmalonicacid,2-hydroxyisocaproicacid,uracil,glutaricacid,adipicacid,2-hydroxyphenylaceticacid,3-hydroxy-3-methylglutaricacid,subericacid,phenypyruvicacid,oroticacid,sebacicacidis8.23
7、min,9.75min,10」8min,12.07min,13.15min,14.79min,15.78min,16.35min,17.60min,17.79min,18.38min,20.16min.Exceptphenypyruvicacid,correlationcoefficientsofalltheotherorganicacidsare0.981-0.999,intra-andinter-assayimprecisionsarelessthan15%,recoveriesrangefrom85%-111%-Afterthe
8、extractionandderivatization,correlationcoefficientofpheypyruvicacidis0.998,intra・andinter-assayimprecisionisle
