Determination of folate vitamers in human serum by stable-isotope-dilution tandem mass spectrometry and comparison with radioassay and microbiologic assay
Determination of folate vitamers in human serum by stable-isotope-dilution tandem mass spectrometry and comparison with radioassay and microbiologic assay
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DOI：
10.1373/clinchem.2003.026955
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发表时间：
2004-02-01
影响因子：
9.3
通讯作者：
Gunter, EW
中科院分区：
文献类型：
--
作者：
Pfeiffer, CM;Fazili, Z;Gunter, EW
背景：目前的临床方法对叶酸的测定结果不同，不能测量各种形式的叶酸。我们建立了一种液相色谱（LC/MS/MS）耦合的同位素稀释串联质谱方法，作为测定人血清中5-甲基四氢叶酸（5MeTHF）、5-甲酰四氢叶酸（5FoTHF）和叶酸（FA）的候选参比方法。方法：采用苯基固相萃取试剂盒从275 muL血清中定量分离叶酸，然后在稳定的血清提取物中采用正离子电喷雾液相色谱/质谱联用技术进行检测和定量。我们在C-8分析柱上采用有机溶剂醋酸等密度流动相。用c -13标记的叶酸作为内标。结果：血清的检出限分别为0.13 （5MeTHF）、0.05 （5FoTHF）、0.07 (FA) nmol/L。运行内和运行间不精确（CV）
Background: Current clinical methods for folate give different results and cannot measure the various forms of folate. We developed an isotope-dilution tandem mass spectrometric method coupled to liquid chromatography (LC/MS/MS) as a candidate reference method for 5-methyltetrahydrofolic acid (5MeTHF), 5-formyltetrahydrofolic acid (5FoTHF), and folic acid (FA) in human serum.Methods: We quantitatively isolated folates from 275 muL of serum with a phenyl solid-phase extraction cartridge, then detected and quantified them in stabilized serum extracts by positive-ion electrospray ionization LC/MS/MS. We used an isocratic mobile phase of acetic acid in organic solvent on a C-8 analytical column. C-13-labeled folates were used as internal standards.Results: Limits of detection in serum were 0.13 (5MeTHF), 0.05 (5FoTHF), and 0.07 (FA) nmol/L. Within- and between-run imprecision (CV) was