HPLC法检测蝉拟青霉中N6-(2-羟乙基)-腺苷含量  

Determination of N6-(2-hydroxyethyl)-adenosine in Paecilomyces cicadae by HPLC

作　　者：李慧雯 艾仁丽 谭艾娟[2] LI Hui-wen;AI Ren-li;TAN Ai-juan(Brand Building Service Station,Rural Revitalization Service Center,Guiyang Bureau of Agriculture and Rural Affairs,Guiyang 550009,China;College of Life Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵阳市农业农村局乡村振兴服务中心品牌建设服务站,贵阳550009 [2]贵州大学生命科学学院,贵阳550025

出　　处：《湖北农业科学》2022年第19期114-118,共5页Hubei Agricultural Sciences

基　　金：贵州省科技计划项目(黔科合平台人才【2017】5788号)。

摘　　要：为了建立蝉拟青霉(Paecilomyces cicadae)5704s中N6-(2-羟乙基)-腺苷(HEA)含量测定方法,以去离子水为溶剂,采用超声辅助水浴法提取HEA,基于HPLC法测定其含量。经优化确定色谱条件为Agilent ZORBAX Extend-C色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇∶5 mmol/L乙酸铵(含0.1%甲酸),梯度洗脱,检测波长为260 nm,流速为1.0 mL/min,柱温为25℃,进样量为10μL。结果显示,HEA浓度为0.25~100.00μg/mL时,峰面积和HEA含量之间线性关系良好,R^(2)=0.9999,仪器的检出限为0.25μg/mL,平均回收率为97.73%~108.95%,RSD为0.37%~1.29%,测得样品中HEA含量为0.78 mg/g。该方法简便、快捷、准确、稳定,适用于蝉拟青霉5704s菌丝体中HEA的测定。In order to establish a method for the determination of N6-(2-hydroxyethyl)-adenosine(HEA)in Paecilomyces cicadae 5704 s,HEA was extracted by ultrasonic assisted water bath method with deionized water as the solvent,and its content was determined by HPLC.The chromatographic conditions were optimized as follows:Agilent ZORBAX Extend-Ccolumn(250 mm×4.6mm,5μm);mobile phase was methanol∶5 mmol/L ammonium acetate(containing 0.1%formic acid);gradient elution;detection wavelength was 260 nm;flow rate was 1.0 mL/min;column temperature was 25℃;injection volume was 10μL.The results showed that when the concentration of HEA was 0.25~100.00μg/mL,the linear relationship between peak area and HEA content was good,R^(2)=0.9999,the detection limit of the instrument was 0.25μg/mL,the average recovery was 97.73%~108.95%,RSD was 0.37%~1.29%,and the content of HEA in the sample was 0.78 mg/g.The method is simple,rapid,accurate,stable and suitable for the determination of HEA in Paecilomyces cicadae 5704s mycelium.

关 键 词：蝉拟青霉(Paecilomyces cicadae) N6-(2-羟乙基)-腺苷(HEA) HPLC 检测 

分 类 号：R284.2[医药卫生—中药学]