增强剂在硫醇的高效薄层色谱荧光分析中的应用。
Use of enhancers in the HPTLC fluorescence analysis of thiols.
作者信息
Ling B L, Baeyens W R, Del Castillo B, Stragier K, Marysael H, De Moerloose P
机构信息
Laboratory of Pharmaceutical Chemistry and Drug Analysis, Faculty of Pharmaceutical Sciences, State University of Ghent, Belgium.
出版信息
J Pharm Biomed Anal. 1989;7(12):1671-8. doi: 10.1016/0731-7085(89)80181-5.
Several thiols of biological and pharmacological interest, including glutathione, coenzyme A, acetylcysteine and captopril were derivatized with the fluorogenic reagents SBD-F and ABD-F and analysed by high-performance thin-layer chromatography (HPTLC)-fluorodensitometry on silica gel 60 plates, using isopropyl ether-methanol-water-acetic acid (9:8:2:1, v/v/v/v) as the developing solvent. The luminescence was considerably increased when several types of enhancers were applied as dipping reagents: Triton X-100, liquid paraffin and cyclodextrins; thus the detectability of the thiol fluorophores was improved. The influence of enhancer concentration, method of application, sample concentration, drying conditions and measuring time after plate dipping were investigated. The greatest enhancement was achieved using a 40% (v/v) solution of Triton X-100 in toluene as a dipping reagent for the determination of SBD-acetylcysteine; more than a 10-fold increase of the fluorescence signal was obtained, allowing low picogram detection limits.
几种具有生物学和药理学意义的硫醇，包括谷胱甘肽、辅酶A、乙酰半胱氨酸和卡托普利，用荧光试剂SBD - F和ABD - F进行衍生化，并在硅胶60板上采用高效薄层色谱（HPTLC）-荧光密度法进行分析，以异丙醚 - 甲醇 - 水 - 乙酸（9:8:2:1，v/v/v/v）作为展开溶剂。当几种类型的增强剂用作浸渍试剂时，发光显著增强：Triton X - 100、液体石蜡和环糊精；因此提高了硫醇荧光团的可检测性。研究了增强剂浓度、应用方法、样品浓度、干燥条件和板浸渍后的测量时间的影响。使用40%（v/v）的Triton X - 100甲苯溶液作为浸渍试剂测定SBD - 乙酰半胱氨酸时，实现了最大程度的增强；获得了超过10倍的荧光信号增强，从而实现了低皮克检测限。
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