蛹虫草培养残基中虫草素的水热回流提取及含量测定

doi:10.16431/j.cnki.1671-7236.2022.09.007

摘要/Abstract

摘要： 【目的】探究水热回流法提取蛹虫草培养残基中活性成分虫草素的最佳工艺参数。【方法】选取液料比、提取次数、提取温度、提取时间4个因素进行单因素试验和正交试验，并通过高效液相色谱法（HPLC）测定虫草素含量，确定水热回流法提取蛹虫草培养残基的最佳工艺参数。【结果】方法学验证结果表明，蛹虫草培养残基中虫草素含量的HPLC测定方法精密度、重复性、稳定性相对标准偏差（RSD）均<2%，准确度RSD为2.64%，标准曲线线性相关性高。正交试验结果显示，影响水热回流法提取效果的因素依次为提取时间、提取次数、液料比、提取温度，其中提取时间和提取次数对提取物虫草素含量均有显著影响（P<0.05），提取温度和液料比影响不明显（P>0.05）。单因素试验和正交试验结果表明，蛹虫草培养残基的最佳提取工艺参数为：液料比25：1、提取次数5次、提取温度70 ℃、提取时间60 min，该提取条件下测得蛹虫草培养残基中虫草素含量为1.48 mg/g。【结论】采用本研究确定的水热回流法最佳工艺参数能有效提取蛹虫草培养基活性成分，且操作简单实用，提取物活性成分得率稳定，为后续蛹虫草培养残基产品开发提供参考依据。

关键词: 蛹虫草培养残基; 虫草素; 水热回流法; 提取工艺

Abstract: 【Objective】 The purpose of this study was to explore the optimal extraction conditions of cordycepin from Cordyceps militaris residual medium by the hydrothermal reflux method.【Method】 The single factor design and the orthogonal experiment design were conducted according to four factors including the liquid to material ratio, extraction repetitions, extraction temperature and extraction time, and the cordycepin content was measured by high-performance liquid chromatography (HPLC) method, which were used to decide the optimal extraction parameters of cordycepin from Cordyceps militaris residual medium by the hydrothermal reflux method.【Result】 The results of methodology showed that the relative standard deviation (RSD) of precision, repeatability, and stability for HPLC detection of cordycepin content in the extract of Cordyceps militaris residual medium were less than 2%, the RSD of accuracy was 2.64%, and the standard curve had high linear correlation.The results of the orthogonal experiment showed that the order of factors affecting the extraction efficacy of the hydrothermal reflux method were extraction time, extraction repetitions, liquid to material ratio and extraction temperature, and extraction time and extraction repetitions had significant effects on cordycepin content in extract (P<0.05), while extraction temperature and liquid to material ratio had not obvious effects (P>0.05).The results of the single factors and the orthogonal experiment design showed that the optimal extraction process factors of cordycepin from Cordyceps militaris residual medium were the ratio of liquid to material 25:1, repetitions 5 times, extraction temperature 70 ℃, and extraction time 60 min, respectively.Under these extraction conditions, the cordycepin content of Cordyceps militaris residual medium was 1.48 mg/g.【Conclusion】 The optimal extraction parameters of the hydrothermal reflux method in this study could effectively extract the active components from Cordyceps militaris residual medium, the operation process was easy and practical, and the yield of the active components of the extract was stable.The results provided reference for the development of product from Cordyceps militaris residual medium.

Key words: Cordyceps militaris residual medium; cordycepin; hydrothermal reflux method; extraction process

中图分类号:

R284.2

彭志苹, 张景艳, 郭志廷, 王磊, 张凯, 张康, 王贵波, 仇正英, 王学智, 李建喜. 蛹虫草培养残基中虫草素的水热回流提取及含量测定[J]. 中国畜牧兽医, 2022, 49(9): 3333-3342.

PENG Zhiping, ZHANG Jingyan, GUO Zhiting, WANG Lei, ZHANG Kai, ZHANG Kang, WANG Guibo, QIU Zhengying, WANG Xuezhi, LI Jianxi. Extraction and Content Detection of Cordycepin from Cordyceps militaris Residual Medium by Hydrothermal Reflux Method[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3333-3342.

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