Quantification of intracellular homocysteine by stable isotope dilution liquid chromatography/tandem mass spectrometry
作者： Yuehua Huang , Zhi-Yong Lu , Karen S. Brown , Alexander S. Whitehead , Ian A. Blair
DOI: 10.1002/BMC.735
关键词:
摘要: A precise and accurate stable isotope dilution liquid chromatography/tandem mass spectrometry method for the analysis of intracellular homocysteine has been developed. An internal standard, [(2)H(8)]-homocystine, was added to cell pellets from EA.hy 926 cells grown in culture under low high folate concentrations. D,L-dithiothreitol used reduce cellular homocystine homocysteine. Cellular proteins were precipitated by addition formic acid acetonitrile. After centrifugation, a portion supernatant analyzed spectrometry. Using Supelcosil cyano column with an Applied Biosystems API 4000 triple quadrupole spectrometer, SRM transitions (m/z 136 m/z 90) [(2)H(4)]-homocysteine 140 94) monitored. The validated conducting five replicate analyses on three different days at four concentrations (concentrations lower limit quantitation expected quartile, mid-range upper quartile). detection 2 ng/10(6) cells. this method, concentration ranged 10 36
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