Shea M, Howell S
Anal Biochem. 1984 Aug 1;140(2):589-94. doi: 10.1016/0003-2697(84)90211-2.
A simple technique using reverse-phase ion-pair liquid chromatography for measurement of exogenous thiosulfate is described. Accurate measurement of thiosulfate in plasma and urine was permitted by precolumn derivatization with monobromobimane, a substance that readily yields fluorescent compounds upon reaction with a variety of biologically important nucleophiles including glutathione, cysteine, and sulfite. Using an injection volume of 50 microliters, as little as 0.16 nmol of thiosulfate was reliably measured. The interassay precision of the method was reflected by a coefficient of variation of 7.7% while the coefficient of variation for interassay analysis was 2.6%. Recovery of thiosulfate from plasma was 96.9 +/- 3.2% and greater than 98% from urine. The simplicity, sensitivity, and precision of the method make it ideal for the study of thiosulfate and other important nucleophiles in body fluids.
本文描述了一种使用反相离子对液相色谱法测量外源性硫代硫酸盐的简单技术。通过用单溴代双马来酰亚胺进行柱前衍生化，可以准确测量血浆和尿液中的硫代硫酸盐。单溴代双马来酰亚胺是一种与包括谷胱甘肽、半胱氨酸和亚硫酸盐在内的多种重要生物亲核试剂反应后容易生成荧光化合物的物质。进样体积为50微升时，可可靠地测量低至0.16纳摩尔的硫代硫酸盐。该方法的批内精密度以变异系数7.7%体现，而批间分析的变异系数为2.6%。硫代硫酸盐从血浆中的回收率为96.9±3.2%，从尿液中的回收率大于98%。该方法的简单性、灵敏度和精密度使其成为研究体液中硫代硫酸盐和其他重要亲核试剂的理想选择。
