Determination of oxalyl thiolesters, N-oxalylcysteine and N-oxalylcysteamine in biological materials.
Determination of oxalyl thiolesters, N-oxalylcysteine and N-oxalylcysteamine in biological materials.
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生物材料中草酰硫酯、N-草酰半胱氨酸和N-草酰半胱胺的测定。
DOI：
10.1016/0003-2697(91)90556-9
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发表时间：
1991
影响因子：
2.9
通讯作者：
Hamilton,GA
中科院分区：
文献类型：
--
作者：
Skorczynski,SS;Yang,CS;Hamilton,GA
描述了一种用于定量分析生物样品中草酰硫醇酯 (OTE) 的便捷方法，草酰硫醇酯是一类新发现的哺乳动物代谢物。通过这种特殊技术，可以确定样品中所有 OTE 的总浓度。该方法首先将生物材料与半胱胺（2-氨基乙硫醇）或半胱氨酸在将 OTE 定量转化为 N-草酰半胱胺（或 N-草酰半胱氨酸）的条件下反应，然后与一溴代二甲胺反应，得到高荧光衍生物，通过反相离子对色谱法进行分析，以四丁基铵离子作为抗衡离子，N-(2-巯基丙酰基)甘氨酸作为内标。该方法能够在单次 HPLC 进样中检测低至 0.6 pmol 的 N-草酰基化合物的双烷衍生物。该方法的应用发现，不仅OTEs而且N-草酰半胱氨酸和N-草酰半胱胺都是哺乳动物的正常代谢产物。在各种大鼠组织中，OTE 浓度范围高达 65 nmol/g（湿重），N-草酰半胱氨酸浓度约为 10 nmol/g，N-草酰半胱胺浓度为 0–3 nmol/g。
A convenient method is described for the quantitative analysis of oxalyl thiolesters (OTEs), a newly discovered class of mammalian metabolites, in biological samples. By this particular technique the total concentration of all OTEs in the sample is determined. The method involves first reacting the biological material with cysteamine (2-aminoethanethiol) or cysteine under conditions that convert OTEs quantitatively to N-oxalylcysteamine (or N-oxalylcysteine), followed by reaction with monobromobimane to give a highly fluorescent derivative that is analyzed by reversed-phase ion-pair chromatography, with tetrabutylammonium ion as the counterion and N-(2-mercaptopropionyl)glycine as an internal standard. The method is capable of detecting as little as 0.6 pmol of the bimane derivative of the N-oxalyl compound in a single HPLC injection. The application of this method has led to the discovery that not only OTEs but also N-oxalylcysteine and N-oxalylcysteamine are normal mammalian metabolites. In various rat tissues the OTE concentration ranges up to 65 nmol/g (wet wt), the N-oxalylcysteine concentration is approximately 10 nmol/g, and the N-oxalylcysteamine concentration is 0–3 nmol/g.