HPLC法检测蝉拟青霉中N6-（2-羟乙基）-腺苷含量

doi:10.14088/j.cnki.issn0439-8114.2022.19.022

湖北农业科学 ›› 2022, Vol. 61 ›› Issue (19): 114-118.doi: 10.14088/j.cnki.issn0439-8114.2022.19.022

HPLC法检测蝉拟青霉中N6-（2-羟乙基）-腺苷含量

李慧雯¹, 艾仁丽¹, 谭艾娟²

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

收稿日期:2022-08-01 出版日期:2022-10-10 发布日期:2022-11-04
通讯作者: 艾仁丽（1997-）,女,贵州铜仁人,在读硕士研究生,研究方向为生物化学与天然活性产物研究,（电话）18685695771（电子信箱）1565111084@qq.com。
作者简介:李慧雯（1992-）,女,贵州贵阳人,实验师, 主要从事农产品检测检验研究,（电子信箱）646869262@qq.com。
基金资助:
贵州省科技计划项目（黔科合平台人才【2017】5788号）

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

LI Hui-wen¹, AI Ren-li¹, TAN Ai-juan²

1. Brand Building Service Station, Rural Revitalization Service Center, Guiyang Bureau of Agriculture and Rural Affairs, Guiyang 550009,China;
2. College of Life Science, Guizhou University, Guiyang 550025, China

Received:2022-08-01 Online:2022-10-10 Published:2022-11-04

摘要/Abstract

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-C₁₈色谱柱（250 mm×4.6 mm,5 μm）,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10 μL。结果显示,HEA浓度为0.25~100.00 μg/mL时,峰面积和HEA含量之间线性关系良好,R²=0.999 9,仪器的检出限为0.25 μg/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。

关键词: 蝉拟青霉（Paecilomyces cicadae）, N6-（2-羟乙基）-腺苷（HEA）, HPLC, 检测

Abstract: 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-C₁₈ column （250 mm × 4.6 mm, 5 μm）; 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 10 μL. The results showed that when the concentration of HEA was 0.25~100.00 μg/mL, the linear relationship between peak area and HEA content was good, R²= 0.999 9, the detection limit of the instrument was 0.25 μg/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.

Key words: Paecilomyces cicadae, N6-（2-hydroxyethyl）-adenosine（HEA）, HPLC, detection

中图分类号:

R284.2

李慧雯, 艾仁丽, 谭艾娟. HPLC法检测蝉拟青霉中N6-（2-羟乙基）-腺苷含量[J]. 湖北农业科学, 2022, 61(19): 114-118.

LI Hui-wen, AI Ren-li, TAN Ai-juan. Determination of N6-（2-hydroxyethyl）-adenosine in Paecilomyces cicadae by HPLC[J]. HUBEI AGRICULTURAL SCIENCES, 2022, 61(19): 114-118.

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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

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