Kågedal B, Carlsson M, Denneberg T
J Chromatogr. 1986 Aug 2;380(2):301-11. doi: 10.1016/s0378-4347(00)83658-0.
Methods for quantitative analysis of total and non-protein-bound 2-mercaptopropionylglycine (2-MPG) in plasma, and total 2-MPG in urine, have been developed. By reduction of urine, plasma or deproteinized plasma samples with tributylphosphine, 2-MPG is liberated from its disulphides, and after clean-up of the sample, 2-MPG is derivatized with N-(7-dimethylamino-4-methyl-3-coumarinyl)maleimide (DACM). The 2-MPG-DACM derivative is then quantified by high-performance liquid chromatography (HPLC) with fluorimetric detection. Both ion-suppression and ion-pair HPLC gave satisfactory chromatograms. The precision of the methods was satisfactory (coefficient of variation 3.1-5.8%), analytical recovery was quantitative (85-99%) and the two HPLC techniques were well correlated (r = 0.99). Five healthy subjects receiving 500 mg of 2-MPG showed maximal total plasma concentration of 13.8-26.9 mumol/l at 3-5 h after intake, and their non-protein-bound 2-MPG was, at the same time, 62-77% of the total 2-MPG. The urinary excretion was 27.8 +/- 3.8% (mean +/- S.D.) of the given dose, most of it excreted within 12 h after intake.
已开发出用于定量分析血浆中总2-巯基丙酰甘氨酸（2-MPG）和非蛋白结合2-MPG以及尿液中总2-MPG的方法。通过用三丁基膦还原尿液、血浆或脱蛋白血浆样品，2-MPG从其二硫化物中释放出来，样品净化后，2-MPG用N-（7-二甲基氨基-4-甲基-3-香豆素基）马来酰亚胺（DACM）进行衍生化。然后用荧光检测的高效液相色谱（HPLC）对2-MPG-DACM衍生物进行定量。离子抑制HPLC和离子对HPLC均给出了令人满意的色谱图。这些方法的精密度令人满意（变异系数为3.1-5.8%），分析回收率为定量（85-99%），两种HPLC技术相关性良好（r = 0.99）。五名接受500 mg 2-MPG的健康受试者在摄入后3-5小时血浆总浓度最高达到13.8-26.9 μmol/l，同时其非蛋白结合2-MPG占总2-MPG的62-77%。尿排泄量为给药剂量的27.8 +/- 3.8%（平均值 +/- 标准差），大部分在摄入后12小时内排出。
