采用5-甲基-(2-(间碘乙酰氨基苯基)苯并恶唑)通过高效液相色谱法和荧光检测法测定硫醇
Determination of thiol by high-performance liquid chromatography and fluorescence detection with 5-methyl-(2-(m-iodoacetylaminophenyl)benzoxazole.
作者信息
Liang Shu-Cai, Wang Hong, Zhang Zhi-Min, Zhang Hua-Shan
机构信息
Department of Chemistry, Wuhan University, Wuhan, 430072, China.
出版信息
Anal Bioanal Chem. 2005 Mar;381(5):1095-100. doi: 10.1007/s00216-004-3006-2. Epub 2005 Mar 3.
A new high-performance liquid chromatographic (HPLC) method for measuring low molecular weight (LMW) thiol-containing compounds, including cysteine (CysH), glutathione (GSH), N-acetylcysteine (Nac), penicillamine (PA), and 2-mercaptoethanol (2-ME), has been developed by using 5-methyl-(2-(m-iodoacetylaminophenyl)benzoxazole (MIPBO) as fluorescence-labeling reagent. The derivatization and separation conditions have been investigated in detail. Detection limits ranging from 3.5 to 15.0 fmol were achieved for the thiols investigated in a 16 min separation with detection wavelengths 310 and 375 nm for the excitation and emission, respectively. The utility of the proposed method has been validated by measuring CysH in human urine samples.
已开发出一种新的高效液相色谱（HPLC）方法，用于测定含低分子量（LMW）硫醇的化合物，包括半胱氨酸（CysH）、谷胱甘肽（GSH）、N-乙酰半胱氨酸（Nac）、青霉胺（PA）和2-巯基乙醇（2-ME），该方法使用5-甲基-（2-（间碘乙酰氨基苯基）苯并恶唑（MIPBO）作为荧光标记试剂。已详细研究了衍生化和分离条件。在所研究的硫醇中，在16分钟的分离时间内实现了3.5至15.0飞摩尔的检测限，激发和发射的检测波长分别为310和375纳米。通过测定人尿样中的CysH，验证了所提出方法的实用性。
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