Houze P, Gamra S, Madelaine I, Bousquet B, Gourmel B
Laboratoire de Biochimie A, Hôpital Saint-Louis, Paris, France.
J Clin Lab Anal. 2001;15(3):144-53. doi: 10.1002/jcla.1018.
We here describe an ion-exchange high-performance liquid chromatography technique with electrochemical detection for rapid quantification of glutathione, homocysteine, cysteinylglycine, and methionine. The analytical validation of the technique showed within-assay and between-assay coefficients of variation between 3.1 and 4.3%, and 3.7 and 8.6%, respectively. Percentages of recovery for overload and dilution tests were between 87 and 120%. Detection limits were 1 micromol/L for methionine and 0.5 micromol/L for other compounds. There was no interference with any physiological and pharmacological substances possessing a thiol function. Aminothiol concentrations determined in 100 control subjects (50 women and 50 men) showed no age- or sex-rated differences for except for homocysteine which was increased (+ 28%) in oldest subjects of both sexes. In 60 patients at risk (30 with chronic renal failure, 30 with diabetes), homocysteine concentration was significantly increased. No variation in other aminothiols was observed in diabetic subjects. Methionine was decreased and cysteinylglycine was increased in patients with chronic renal failure. The present technique-rapid, easy to use, and reliable-appears suitable for routine application in the exploration of aminothiol metabolic pathways including mechanisms of hyperhomocysteinemia.
我们在此描述一种采用电化学检测的离子交换高效液相色谱技术，用于快速定量谷胱甘肽、同型半胱氨酸、半胱氨酰甘氨酸和蛋氨酸。该技术的分析验证显示批内变异系数和批间变异系数分别在3.1%至4.3%以及3.7%至8.6%之间。过载和稀释试验的回收率在87%至120%之间。蛋氨酸的检测限为1微摩尔/升，其他化合物的检测限为0.5微摩尔/升。对任何具有硫醇功能的生理和药理物质均无干扰。在100名对照受试者（50名女性和50名男性）中测定的氨基硫醇浓度显示，除同型半胱氨酸外，无年龄或性别差异，同型半胱氨酸在两性年龄最大的受试者中升高（+28%）。在60名有风险的患者（30名慢性肾衰竭患者和30名糖尿病患者）中，同型半胱氨酸浓度显著升高。在糖尿病患者中未观察到其他氨基硫醇的变化。慢性肾衰竭患者的蛋氨酸降低，半胱氨酰甘氨酸升高。目前这种快速、易于使用且可靠的技术似乎适用于探索氨基硫醇代谢途径（包括高同型半胱氨酸血症机制）的常规应用。
