柱前衍生化-高效液相色谱法跟踪检测酶法制备γ-D-谷氨酰-L-色氨酸反应过程  

Analysis of γ-D-Glutamyl-L-tryptophan in Enzymatic Synthesis Procedure by High Performance Liquid Chromatography

作　　者：屠春燕[1] 吴燕娇[1] 袁艳娟[1] 姚忠[1] 唐美华[1] 韦萍[1] 

机构地区：[1]南京工业大学生物与制药工程学院,南京210009

出　　处：《分析化学》2009年第10期1528-1530,共3页Chinese Journal of Analytical Chemistry

摘　　要：γ-D-谷氨酰-L-色氨酸(SCV-07)是一种新型广谱免疫调节类化合物。针对酶法合成SCV-07反应体系的特点,以邻硝基苯磺酰氯为柱前衍生试剂,RP-HPLC为分离模式,选用Lichrospher C18柱,20mmol/L磷酸盐溶液(pH3.0)和乙腈为流动相,梯度洗脱,检测波长230nm,建立了同时分析测定多种氨基酸和小肽的方法,所有组分于15min内洗脱完毕。氨基酸及二肽在0.96～260μmol/L范围内线性良好,相关系数大于0.9991;加标回收率在97.8%～101.6%之间;相对标准偏差为1.2%～2.9%。本方法适用于SCV-07产品的纯度分析,以及酶法转肽制备SCV-07反应过程跟踪检测,具有分析速度快,准确度高、操作简单等优点。γ-D-Glutamyl-L-tryptophan（SCV-07） is a prospective medicine for the treatment of tuberculosis.The RP-HPLC method was used to analyze the practical system of SCV-07 by enzymatic synthesis from D-glutamine and L-tryptophan.Under the optimal conditions,the chromatographic conditions were as follows：Lichrospher C18 column（250 mm×4.6 mm,5 μm） was used,the detection wavelength was 230 nm.Flow rate was 1.0 mL/min;mobile phase A was 20 mmol/L sodium dihydrogen phosphate（pH 3.0）,mobile phase B was ACN;the gradient protocol was applied.All compounds were completely eluted in 15 min.The linear relationship was obtained between peak area and concentration for D-Gln,L-Glu,SCV-07 and L-Trp with correlation coefficients higher than 0.9991.The recoveries of the analyses from spiking experiments were 97.8%-101.6%,with relative standard deviations being 1.2%-2.9%.This method is fast,accurate,sensitive,and easy to run.

关 键 词：色氨酸 邻硝基苯磺酰氯 衍生化 反相高效液相色谱 

分 类 号：TQ922.1[轻工技术与工程—发酵工程]