一种提高高半胱氨酸液相色谱检测灵敏度的方法

揭新明;王媚

中国药学杂志 ›› 1999, Vol. 34 ›› Issue (05) : 324-326.

中国药学杂志 ›› 1999, Vol. 34 ›› Issue (05) : 324-326.
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一种提高高半胱氨酸液相色谱检测灵敏度的方法

  • 揭新明;王媚
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A sensitive method for the determination of homocysteine by HPLC

  • Jie Xinming
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摘要

目的:建立一种用高效液相色谱测定微量高半胱氨酸的新方法?方法:反应条件研究采用分子荧光光谱法,高半胱氨酸测定采用反相高效液相色谱法?结果:使用邻苯二甲醛作荧光衍生试剂,在温度为50℃,pH8.4,反应4min时,衍生物相对荧光强度最大?在5×10-9~2.7×10-7mol·L-1浓度范围内,荧光强度与浓度之间呈线性关系,检测限为4.2×10-10mol·L-1?结论:经过氧化氢处理后,HCY与OPA的衍生物荧光强度可提高5倍?

Abstract

OBJECTIVE:To establish a sesitive method to determine homocysteine by a reversed-phase high-performance liquid chromatography (RP-HPLC).METHODS:Homocysteine was determined by RP-HPLC with Fluorescence spectrophotometry.RESULTS:Ophthaladehyde was used as fluorescent derivative reagent.When the reaction took place at 50℃ and pH8.4 for 4 min,the fluorescent intensity reached the highest.In the concentraction of 5×10-9~2.7×10-7mol·L-1,F%=6.52×109c-0.91,r=0.998,RSD=2.24%,detection limit=4.2×10-10mol·L-1.CONCLUSION:Treated with H2O2,the fluorescent intensity of ophthalaldehyde derivatives of HCY may increase by 5 times.

关键词

高半胱氨酸 / 高效液相色谱

Key words

homocyteine / HPLC / cardiovascular disease

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导出引用
揭新明;王媚. 一种提高高半胱氨酸液相色谱检测灵敏度的方法[J]. 中国药学杂志, 1999, 34(05): 324-326
Jie Xinming. A sensitive method for the determination of homocysteine by HPLC[J]. Chinese Pharmaceutical Journal, 1999, 34(05): 324-326

参考文献

1 梁效海,韩毅.HCY代谢与动脉粥样硬化的关系.生命的化学,1993,13(5):30. 2 陈国珍,黄贤智,郑朱梓,等.荧光分析法.北京:科学出版社,1990:56. 3 Chen RF.Flucorescence properties of ophthalaldehyde derivatives of amino acid.Biochem Biophys Acta,1979,576(2):440. 4 Alvarez-Coque MCG.Formation and instability of ophtha-laldehyde derivatives of aminoacid.Anal Biochem,1989,178(1):1. 5 Schram E,MooreS,Bigwood EJ.Chromatographic determination of cystineas cystric acid.J.Biochem,1954,57(1):33. 6 Hirs CHW.Performic acid oxidation.Methods Enzymol,1967,11(1):197. 7 Gurd FRN.Carboxy methylation.Methods Enzymol,1972,25(1):424. 8 Barkholt V.Amino acid analysis:determination of cysteine plus halfcystine in proteins after hydrochloric acid hydrolysis with a disulfide compound as additive.Anal Biochem,1989,177(2):318.

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