摘要
目的 建立人血浆和全血中谷胱甘肽浓度的HPLC-MS测定法。方法 血浆或全血样品用6％高氯酸沉淀蛋白后直接进样测定,以SinoChrom ODS-BP (200mm×4.6mm,5μm)分离,质谱检测器选择m/z 308(谷胱甘肽)进行检测,外标法以峰面积定量。结果 本方法以质谱检测器选择离子检测,内源性杂质不干扰谷胱甘肽的分离检出,血浆或全血农度在0.25～250μg·mL^(-1)范围内,峰面积与浓度线性关系良好,平均回收率为99.6％～101.0％,日内、日间RSD分别≤4.3％和≤4.6％。10名健康志愿者的血浆还原型谷胱甘肽的浓度为(0.50+0.10)μg·mL^(-1),全血中浓度为(171.5±35.2)μg·mL^(-1)。结论 HPLC-MS用于测定生物样品中谷胱甘肽浓度,样品处理简单,方法准确可靠,可用于谷胱甘肽在人血浆和全血中的代谢和转运情况的研究。
OBJECTIVE To establish a HPLC-MS method to determine the reduced glutathione (GSH) in human plasma and blood. METHODS Plasma and blood samples were separated in a SinoChrom ODS-BP (200 mm×4.6 mm, 5μm) after protein precipitated by 6% Operchloric, and m/z 308 was monitored by a micromass ZQ detector. RESULTS Retention time of GSH was about 4 min, and other endogenous in the plasma or blood did not interfere the test results. The calibration curve of GSH was linear within the range of 0.25～250μg·mL^(-1)(r=0.9995), the recovery of this method was 99.6%～101.0%, and the within day and between day RSD were no more than 4.3% and 4.6%, respectively. The concentrations of GSH were (0.50±0.10)μg·mL^(-1) in plasma and (171.5±35.2) μg·mL^(-1) in blood for 10 Chinese male volunteers. CONCLUSIONS HPLC-MS is a simple, reliable method for the determination of GSH in plasma or blood, and is fit for determining the disposition of GSH in human body.
出处
《中南药学》
CAS
2004年第3期154-156,共3页
Central South Pharmacy
