Pastore A, Massoud R, Motti C, Lo Russo A, Fucci G, Cortese C, Federici G
Laboratory of Biochemistry, Children Hospital Bambino Gesù IRCCS, Rome, Italy.
Clin Chem. 1998 Apr;44(4):825-32.
We describe a 6-min HPLC method to measure the total concentrations of the most important thiols in plasma and urine--cysteine, homocysteine, cysteinylglycine, and glutathione--as well as the concentrations in plasma and urine, respectively, of cysteamine and 2-mercaptopropionylglycine, two compounds used to treat disorders of cysteine metabolism. Precolumn derivatization with bromobimane and reversed-phase HPLC were performed automatically by a sample processor. Throughput was up to 100 samples in 24 h. The within-run CV ranged from 0.9% to 3.4% and the between-run CV ranged from 1.5% to 6.1%. Analytical recovery was 97-107%, with little difference between plasma and urine samples. The detection limit was approximately 50 nmol/L for all the analytes studied. Thiol concentrations were determined in the plasma of 206 healthy donors and in the urine of 318 healthy donors distributed for age and sex. Mean values of plasma cysteine and homocysteine were significantly lower in infants (ages, <1 y) compared with other age groups (P <0.005). In adults, mean plasma homocysteine values were higher in males than in females (9.2 vs 6.7 micromol/L, P <0.0001) and in the 6- to 10-year-old group (P <0.05). Mean values for glutathione and cysteinylglycine were not sex- and age-dependent. In urine, both cysteine and homocysteine showed a wide range of variation.
我们描述了一种6分钟的高效液相色谱法，用于测量血浆和尿液中最重要的硫醇（半胱氨酸、同型半胱氨酸、半胱氨酰甘氨酸和谷胱甘肽）的总浓度，以及分别用于治疗半胱氨酸代谢紊乱的两种化合物——半胱胺和2-巯基丙酰甘氨酸在血浆和尿液中的浓度。通过样品处理器自动进行溴代双马来酰亚胺柱前衍生和反相高效液相色谱分析。24小时内通量高达100个样品。批内变异系数在0.9%至3.4%之间，批间变异系数在1.5%至6.1%之间。分析回收率为97%至107%，血浆和尿液样品之间差异不大。所有研究分析物的检测限约为50 nmol/L。测定了206名健康供体血浆和318名按年龄和性别分布的健康供体尿液中的硫醇浓度。与其他年龄组相比，婴儿（年龄<1岁）血浆中半胱氨酸和同型半胱氨酸的平均值显著较低（P<0.0０5）。在成年人中，男性血浆同型半胱氨酸的平均值高于女性（9.2对6.7 μmol/L，P<0.0001），在6至10岁组中也是如此（P<0.05）。谷胱甘肽和半胱氨酰甘氨酸的平均值与性别和年龄无关。在尿液中，半胱氨酸和同型半胱氨酸的变化范围都很大。
