High performance liquid chromatography with fluorimetric detection for the determination of total homocysteine in human plasma: method and clinical applications

被引:36
作者
Chou, ST
Ko, LE
Yang, CS
机构
[1] Providence Univ, Dept Food & Nutr, Taichung 43301, Taiwan
[2] Taichung Vet Gen Hosp, Dept Med Res, Taichung, Taiwan
关键词
HPLC; homocysteine; fluorimetric detection; monobromobimane;
D O I
10.1016/S0003-2670(00)01296-4
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Homocysteine (Hcy), an amino acid derived from methionine is an independent risk factor for coronary artery disease. We have developed a convenient high performance liquid chromatography assay with fluorimetric detection for the accurate determination of Hey in human plasma. This assay involves direct addition of human plasma to methanolic monobromobimane for simultaneous protein precipitation and thiol derivatization. The assay was validated by adding authentic homocysteine and homocystine to plasma samples with recovery rates between 95.5 and 102.4%. The relative standard deviations for intraday (n = 7) and interday (n = 6, 10 consecutive days) analysis were 0.7-3.9 and 0.7-4.0%, respectively. Reference values of plasma total homocysteine (tHcy) have been determined for men and women, showing a significant difference (P < 0.05) between genders. Preliminary results for occluded coronary artery disease and for maintenance hemodialysis are presented. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:331 / 336
页数:6
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