测定血浆总同型半胱氨酸水平的高效液相色谱法

High-performance liquid chromatographic method for measuring total plasma homocysteine levels.

作者信息

Fermo I, Arcelloni C, Mazzola G, D'Angelo A, Paroni R

机构信息

Laboratory of Chromatographic and Separative Techniques, IRCCS H San Raffaele, Milan, Italy.

出版信息

J Chromatogr B Biomed Sci Appl. 1998 Nov 20;719(1-2):31-6. doi: 10.1016/s0378-4347(98)00405-8.

DOI:10.1016/s0378-4347(98)00405-8

PMID:9869361

Abstract

We have modified a high-performance liquid chromatographic (HPLC) procedure based on SBD-F (ammonium-7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate) pre-column derivatization to obtain an assay that is useful for routine clinical total plasma homocysteine (tHcy) analysis. The introduction of easily handled sodium borohydride instead of the traditional tri-n-butylphosphine in dimethylformamide as a reductant and a 14-min run-time using basic isocratic HPLC equipment are the more notable advantages. The addition of mercaptopropionylglycine as an internal standard contributed to improvements in the reproducibility of the assay, yielding within- and between-run precisions of 1.9 and 4% (C.V.), respectively. Reference values for fasting tHcy were 7.65+/-2.3 and 8.9+/-2.4 micromol/l, while post-methionine load gave tHcy levels of 19.9+/-5.5 and 26.8+/-5.5 micromol/l, for women and men, respectively (n=40).

摘要

我们改进了一种基于SBD-F（7-氟苯并-2-恶唑-1,3-二氮杂环丁烷-4-磺酸铵）柱前衍生化的高效液相色谱（HPLC）方法，以获得一种可用于常规临床总血浆同型半胱氨酸（tHcy）分析的检测方法。采用易于操作的硼氢化钠代替传统的三正丁基膦在二甲基甲酰胺中作为还原剂，以及使用碱性等度HPLC设备实现14分钟的运行时间，是较为显著的优点。添加巯基丙酰甘氨酸作为内标有助于提高检测的重现性，批内和批间精密度分别为1.9%和4%（变异系数）。女性和男性空腹tHcy的参考值分别为7.65±2.3和8.9±2.4微摩尔/升，而蛋氨酸负荷后tHcy水平分别为19.9±5.5和26.8±5.5微摩尔/升（n = 40）。