Speciation and quantitation of underivatized and Ellman's derivatized biological thiols and disulfides by capillary electrophoresis
Speciation and quantitation of underivatized and Ellman's derivatized biological thiols and disulfides by capillary electrophoresis
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DOI：
10.1006/abio.1996.0439
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发表时间：
1996-11-01
影响因子：
2.9
通讯作者：
Rabenstein, DL
中科院分区：
文献类型：
--
作者：
Russell, J;Rabenstein, DL
已经开发了毛细管电泳（CE）方法用于生物学感兴趣的硫醇和二硫化物的形态和定量，包括内源性化合物谷胱甘肽、谷胱甘肽二硫化物、半胱氨酸、胱氨酸、高半胱氨酸和高胱氨酸以及治疗剂青霉胺、青霉胺二硫化物、N-乙酰半胱氨酸和卡托普利，使用200 nm的检测波长，未衍生化的硫醇和二硫化物实现了良好的形态和定量;使用50 μ m直径的毛细管，检测限在20-90 μ M（1-4 pmol）的范围内。为了达到较低的检测限，硫醇衍生与硫醇特异性探针分子，5，5 '-二硫代-双-（2-硝基苯甲酸）（埃尔曼试剂），良好的形态和定量埃尔曼的衍生硫醇实现使用357 nm的检测波长;检测限范围为5-50 μ M（0.03-0.3 pmol）使用25 μ m直径的毛细管。将未衍生化和衍生化方法应用于人红细胞中谷胱甘肽的测定。对于分析的红细胞样品，获得了2-3 nM的谷氨酸浓度，通过两种方法获得的结果之间具有良好的一致性。(C)出版社：Academic Press，Inc.
Capillary electrophoresis (CE) methods have been developed for the speciation and quantitation of thiols and disulfides of biological interest, including the endogenous compounds glutathione, glutathione disulfide, cysteine, cystine, homocysteine, and homocystine and the therapeutic agents penicillamine, penicillamine disulfide, N-acetylcysteine, and captopril, Good speciation and quantitation were achieved for the underivatized thiols and disulfides using a detection wavelength of 200 nm; detection limits were in the range 20-90 mu M (1-4 pmol) using a 50-mu m-diameter capillary. To achieve lower detection limits, thiols were derivatized with the thiol-specific probe molecule, 5,5'-dithio-bis-(2-nitrobenzoic acid) (Ellman's reagent), Good speciation and quantitation were achieved for the Ellman's derivatized thiols using a detection wavelength of 357 nm; detection Limits were in the range 5-50 mu M (0.03-0.3 pmol) using a 25-mu m-diameter capillary. Both the underivatized and derivatized methods were applied to the determination of glutathione in human erythrocytes. Glutathione concentrations of 2-3 nM were obtained for the erythrocyte samples analyzed, with good agreement between results obtained by the two methods. (C) 1996 Academic Press, Inc.