Simultaneous Quantification of Homocysteine and Folate in Human Serum or Plasma Using Liquid Chromatography/Tandem Mass Spectrometry
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- 作者：Bryant C. Nelson ; Mary B. Satterfield ; Lorna T. Sniegoski ; and Michael J. Welch
- 刊名：Analytical Chemistry
- 出版年：2005
- 出版时间：June 1, 2005
- 年：2005
- 卷：77
- 期：11
- 页码：3586 - 3593
- 全文大小：104K
- 年卷期：v.77,no.11(June 1, 2005)
- ISSN：1520-6882
文摘
A unified extraction and quantification procedure basedon stable isotope-dilution liquid chromatography/tandemmass spectrometry (LC/MS/MS) has been developed forthe simultaneous determination of total homocysteine andfolate (5-methyltetrahydrofolic acid and folic acid) levelsin human serum and plasma. This is the first reportdocumenting the simultaneous extraction and quantification of these structurally dissimilar analytes. Analytes arequantitatively isolated from samples (500 
tities/mgr.gif">L) prior to LC/MS/MS analysis using a two-step stabilization processcombined with C18 solid-phase extraction. The methodexhibits excellent linearity over 4 orders of magnitude foreach analyte. Measurement repeatability (RSD, N = 2)ranged from 0.3% to 3% for all analytes over 1 day ofanalysis. Total method variability (RSD, N = 6) rangedfrom 0.7% to 10% for all analytes over three independentdays of analysis. The accuracy and practical applicabilityof the method were demonstrated by applying the methodto the quantitative determination of each analyte in a newNIST serum Standard Reference Material (NIST SRM1955 Homocysteine and Folate in Frozen Human Serum)and in a small subset of normal donor plasma samples.
tities/mgr.gif">L) prior to LC/MS/MS analysis using a two-step stabilization processcombined with C18 solid-phase extraction. The methodexhibits excellent linearity over 4 orders of magnitude foreach analyte. Measurement repeatability (RSD, N = 2)ranged from 0.3% to 3% for all analytes over 1 day ofanalysis. Total method variability (RSD, N = 6) rangedfrom 0.7% to 10% for all analytes over three independentdays of analysis. The accuracy and practical applicabilityof the method were demonstrated by applying the methodto the quantitative determination of each analyte in a newNIST serum Standard Reference Material (NIST SRM1955 Homocysteine and Folate in Frozen Human Serum)and in a small subset of normal donor plasma samples.