HPLC方法测定生物样品中谷胱甘肽及其构成氨基酸  

Deteminations of reduced glutathion and its component aminoacids in biosample by HPLC

作　　者：石京山[1] 李峰[2] 盛美萍[2] 江文德[2] 

机构地区：[1]遵义医学院药理学教研室 [2]上海医科大学基础医学院药理学教研室

出　　处：《遵义医学院学报》1998年第3期1-4,共4页Journal of Zunyi Medical University

摘　　要：目的建立GSH及其三种构成氨基酸的HPLC分析方法。方法GSH及其构成氨基酸用丹磺酰氯荧光衍化，采用单泵非梯度洗脱，荧光检测器测定，激发波长328um，发射波长541nm。结果各待测成份分离良好，HPLC色谱图清晰，峰整齐对称，血浆中GSH、Glu和Gly的最低检测限O．5nmol／ml，Cys的最低检测限为1．onmol／ml。衍化产物稳定72h以上不衰变，GSH、Glu、Gys及Gly的回收率分别为87.2％、87.4％、82．2％和97．2％，精密度为0.5％－3．8％。经测定Beagle犬血浆GSH、Glu及Gly的正常值分别为29．0nmol/ml、22．0nmol／ml和91．8／nmol/ml，而Cys的生理值在检测限以下。结论本法测定GSH及其构成氨基酸简便、稳定、灵敏。To determined reduced glutahione (GSH ), glumate (Glu), cysteine (Cys ), and glycine(Gly) by high-peifortnance liquid chromatographic(HPLC) method utilizing variable wavelength fluoremetricdetector. Methods A precolum derivatization of etandards and samples was carried out with dansyl chlotide andthe dansyl derivatives wre well separated on C18-ODS column with a methanol-acetate ungradient elutionsolution simultaneously. The fluorescence detector was set to the excitation and emission wavelengths of 328 and541nm, respechvely. ResultS An optimal purity chromatographic peaks were obtained and the lowestdetermined levels of GSH, Glu and Gly were 0 .5nmol/nil(Cys as 1 .0nmol/ml). The recovery rates of GSH,Glu,Cys and Cly in the Plasma of dog were 87. 2%, 87. 4%, 82. 2% and 97. 2% respectively. The overallcoefficient of variation was between 0. 5% ~ 3. 8%. The stability of the derivatives was for more than 72h at4℃. The normal physiological GSH, Glua and Gly concentrations in plasma of beagle dog were 29. 0nmol/ml,22 .0nmol/ml and 91. 8nmol/ml. However, the level of Cys was less than the lowest detector sensitivity.Conlusions present method determined GSH and its Composition andnoacids was sample, stamble andsensitive.

关 键 词：谷胱甘肽 氨基酸 高效液相色谱 血浆 

分 类 号：R446.112[医药卫生—诊断学]