Indirect chiral separation of tryptophan enantiomers by high performance liquid chromatography with indirect chemiluminiscence detection
【作者】 Jie Zhou, Shanshan Chen, Fang Sun, Pei Luo, Qiuzheng Du, Suzhen Zhao
【期刊名】 Journal of Chromatography B
【影响因子】 2015年: 2.687
【作者单位】 School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, PR China
【年，卷(期)：页码】2015, 1006 (2015) 65–70
【关键词】chiral separation; high performance liquid chromatography
【摘要】In recent years, the study of chiral compounds in vivo has received much attention. In this study, a novel method based on high performance liquid chromatography (HPLC) coupled with chemiluminescence (CL) detection was developed for the separation of tryptophan (Trp) enantiomers. o-Phthalaldehyde and N-acetyl-l-cysteine were used as chiral derivatization reagents for Trp before it can be detected by HPLC-CL method. The separation was carried out on an ODS column using a mobile phase com-posed of methanol—0.01 mol/L phosphate buffer (40/60, v/v). Under the optimum conditions, satisfactory results were obtained, including complete separation, good relative standard deviations and low detec-tion limits. The applicability of the proposed method has been validated by determining Trp in biological samples. Linear responses (r > 0.9990) were observed over the range of 2.5 × 10−7 to 1.2 × 10−5 g/mL of Trp enantiomers, with quantitation limit of 2.5 × 10−7 g/mL. The assay method shows good specificity to Trp enantiomers, and thus it will have great potential application in clinical diagnosis. The mean extrac-tion efficiency of Trp enantiomers in mice plasma samples were 98.48% and 97.40%, respectively. The mean relative standard deviation (RSD) of Trp enantiomers were <3%.
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