Characterization of lysophosphatidic acid subspecies produced by autotaxin using a modified HPLC ESI-MS/MS method
[摘要] Lysophosphatidicacid(LPA)isabioactivelipidwithaplethoraofbiologicalfunctionsincludingrolesincellsurvival,proliferation,andmigration.Althoughhigh-performanceliquidchromatographyelectrosprayionizationtandemmassspectrometry(HPLCESI-MS/MS)technologyhasbeenusedtomeasurethelevelsofLPAinhumanblood,serumandplasma,currentmethodscannotreadilydetecttheminutelevelsofLPAfromcellculture.Inthisstudy,amodifiedHPLCESI-MS/MSmethodwithenhancedsensitivitywasdeveloped,whichallowsaccuratemeasurementsofLPAlevelswithalimitofquantitationatapproximately10femtomoles.ThemethodwasvalidatedbyquantitationofLPAlevelsinthemediaofpreviouslycharacterizedcelllinesectopicallyexpressingautotaxin.Specifically,autotaxinoverexpressioninducedanincreaseinthe16:0,18:2,18:1,18:0,and20:4subspeciesofLPA,butnotthe22:6LPAsubspecies.Lastly,thisHPLCESI-MS/MSmethodwascross-validatedviabiologicalassayspreviouslyutilizedtoassayLPAlevels.Hence,thisHPLCESI-MS/MSmethodwillallowresearcherstomeasureinvitroLPAlevelsandalsodistinguishbetweenspecificLPAsubspeciesforthedelineationofindividualbiologicalmechanisms...
[发布日期] [发布机构]
[效力级别] [学科分类] 分析化学
[关键词] [时效性]
