水蛭素的HPLC色谱分析条件优化研究  

Optimization of HPLC Conditions for Analysis of Hirudin

作　　者：黄伟 邓斌[2] 王议娴[3] 陈康 饶荣通[3] 杨秀娟 HUANG Wei;DENG Bin;WANG Yixian;CHEN Kang;RAO Rongtong;YANG Xiujuan(College of Agronomy and Life Sciences,Kunming University,Kunming 650214;Yunnan Rural Science and Technology Service Center,Kunming 650224;Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201;Yunnan Key Laboratory of Animal Nutrition and Feed Science,Kunming 650201)

机构地区：[1]昆明学院农学与生命科学学院,昆明650214 [2]云南省农村科技服务中心,昆明650224 [3]云南农业大学动物科学技术学院,昆明650201 [4]云南省动物营养与饲料重点实验室,昆明650201

出　　处：《中国农学通报》2024年第11期142-147,共6页Chinese Agricultural Science Bulletin

基　　金：云南省重大科技专项计划“药用动物源性中药材质量控制体系建设”(2018ZF012)。

摘　　要：为研究水蛭素的HPLC最佳色谱分析条件,通过对比重组水蛭素的HPLC分析方法的洗脱条件、检测波长、色谱柱的分离效果,拟合水蛭素浓度与峰面积的线性方程,优选最佳水蛭素色谱分析条件。结果显示,ODSC18柱分析最佳条件为:采用梯度洗脱,流动相A和B洗脱程序:0~60 min,B:0%~100%,进样量20μL,柱温35℃,流速为1 mL/min,波长205 nm;TSKgelG2000SW柱最佳分析条件为:流动相磷酸缓冲液(0.02 mol/L,pH 7.0),流速0.5 mL/min,检测波长205 nm,进样量20μL。研究表明,水蛭素浓度与峰面积具有良好的直线线性相关关系,采用TSKgelG2000SW色谱柱的分析效果优于ODSC18柱,可为动物活性多肽的分析提供依据。In order to study the optimal chromatographic conditions for the HPLC analysis of hirudin,we compared the elution conditions,detection wavelengths and separation effects of the HPLC methods for recombinant hirudin,fitted the linear equations of hirudin concentration and peak area,and optimized the hirudin chromatographic analysis conditions.The results showed that the optimal conditions for the analysis on the ODSC18 column were as follows:gradient elution,mobile phases A and B elution program:0~60 min,B:0%~100%,injection volume of 20μL,column temperature of 35℃,flow rate of 1 mL/min,and wavelength of 205 nm.And the optimal conditions for the analysis on the TSKgelG2000SW column were as follows:mobile phase’s phosphate buffer(0.02 mol/L,pH 7.0),flow rate of 0.5 mL/min,and wavelength of 205 nm,injection volume of 20μL.The study showed that the concentration of hirudin had a good linear correlation with the peak area,and the analytical effect of the TSKgelG2000SW column was better than that of the ODSC18 column,which could be used for the analysis of the active peptides in animals.

关 键 词：水蛭素 液相色谱 色谱柱 色谱条件 分离效果 

分 类 号：S185[农业科学—农业基础科学]