Analysis of plasma thiols by high-performance liquid chromatography with ultraviolet detection
Analysis of plasma thiols by high-performance liquid chromatography with ultraviolet detection
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DOI：
10.1016/j.chroma.2003.11.030
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发表时间：
2004-04-02
影响因子：
4.1
通讯作者：
Kusmierek, K
中科院分区：
文献类型：
--
作者：
Bald, E;Chwatko, G;Kusmierek, K
本文提出了测定血浆中四种不同形式巯基半胱氨酸、半胱氨酰甘氨酸、谷胱甘肽和同型半胱氨酸的方法。用硫醇特异性紫外线标记试剂2-氯-1-甲基喹啉四氟硼酸盐对分析物进行衍生化，并通过反相高效液相色谱法在355 nm处检测，将分析物彼此、试剂过量和血浆基质成分分离。在衍生化步骤之前，通过用硼氢化钠还原裂解将氧化形式转化为其硫醇对应物。为了避免样品制备过程中由于氧化而导致的硫醇还原部分的损失，在采集后和血浆与红细胞分离前立即向全血中加入衍生试剂。该方法测量总硫醇、总游离硫醇、蛋白结合硫醇和还原硫醇。该方法在硫醇的生理和病理范围内呈线性，并适用于表面健康志愿者捐赠的血浆样本。(C)2004年爱思唯尔B. V.保留所有战斗。
The procedure for measurement of different forms of four plasma thiols cysteine, cysteinylglycine, glutathione and homocysteine is proposed. The analytes are derivatized with thiol-specific ultraviolet labeling reagent, 2-chloro-1-methylquinolinium tetrafluoroborate, and separated from each other, reagent excess and plasma matrix constituents by reversed-phase high performance liquid chromatography with detection at 355 nm. Oxidized forms are converted to their thiol counterparts by reductive cleavage with sodium borohydride prior to derivatization step. In order to circumvent the loss of reduced fraction of thiols due to oxidation during sample preparation, the derivatization reagent is added to whole blood immediately after collection and before separation of plasma from erythrocytes. The method measures total thiols, total free thiols, protein-bound thiols and reduced thiols. The method is linear within the physiological and pathological ranges of thiols and is applied for plasma samples donated by apparently healthy volunteers. (C) 2004 Elsevier B.V. All fights reserved.