Analysis of thiols by HPLC after fluorescent prelabelling with 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid |
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Authors: | V. Cavrini R. Gatti P. Roveri A. Di Pietra |
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Affiliation: | (1) Department of Pharmaceutical Sciences, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy |
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Abstract: | Summary The use of 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid as a fluorogenic reagent in pre-column derivatization for the high-performance liquid chromatography (HPLC) of biologically important thiols (L-cysteine, glutathione, N-acetylcysteine, homocysteine and mercaptopropionylglycine) was investigated. The aroylacrylic acid reacts selectively and rapidly (15 min. at room temperature) with the thiol compounds to give stable fluorescent adducts which can be separated by reversedphase HPLC and detected fluorometrically (ex 300nm); em 445nm). The experimental conditions for the thiol derivatization and chromatographic separation are discussed. Applications to the determination of N-acetylcysteine, mercaptopropionylglycine and cysteine are described.Presented in part at Bononiachem, XVI Congresso Nazionale di Chimica, Bologna, October 9–14, 1988. |
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Keywords: | Column liquid chromatography Thiol analysis Fluorimetric detection Derivatization with 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid |
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