Myung S W, Kim M, Min H K, Yoo E A, Kim K R
Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology, Cheongryang, Seoul, South Korea.
J Chromatogr B Biomed Sci Appl. 1999 Apr 30;727(1-2):1-8. doi: 10.1016/s0378-4347(99)00071-7.
The purpose of this study was to develop a simple and accurate analytical method to determine homocysteine (Hcy), cysteine (Cys), and methionine (Met) in aqueous samples. Until now, the most frequently used method for the assay of Hcy, Cys, and Met has been high-performance liquid chromatography with fluorescence detection after fluorescent tagging. The newly developed method involves the employment of the SPME (solid-phase microextraction) technique together with GC-MS. For application to a gas chromatographic system, alkyl formate derivatives were prepared in the form of N(O,S)-alkoxycarbonyl alkyl ester with the analytes in the aqueous samples. The optimum derivatizing regent for N(O,S)-alkoxycarbonyl alkyl ester was chosen by comparing the efficiency of the derivatized analytes in a GC through the SPME method and liquid-liquid extraction. The optimum conditions of the SPME system for the analytes derivatized with N(O,S)-ethoxycarbonyl propyl ester in the aqueous matrix were pH 3.0 and no salt, and 30 min equilibration time using an 85 microm PA (polyacrylate) fiber. The developed method is inexpensive, easy and rapid.
