食品科学 ›› 2011, Vol. 32 ›› Issue (5 ): 239-243.doi: 10.7506/spkx1002-6630-201105051

• 生物工程 • 上一篇    下一篇

微生物发酵产大豆异黄酮苷元的连续超声波提取工艺及其定量分析研究

张晓玲,惠芸华,杨 桥*   

  1. 中国水产科学研究院东海水产研究所,农业部水产品质量监督检验测试中心
  • 收稿日期:2010-09-25 修回日期:2011-02-14 出版日期:2011-03-15 发布日期:2011-03-03
  • 通讯作者: 杨桥 E-mail:qyang@mail.whu.edu.cn
  • 基金资助:
    中央公益性科研院所基本科研业务费专项资金资助项目(2008M17);上海市自然科学基金项目(11ZR1450000)

Continuous Ultrasonic Extraction and Quantitative Analysis of Isoflavone Aglycones Produced by  Microbial Fermentation

ZHANG Xiao-ling,HUI Yun-hua,YANG Qiao*   

  1. Supervision and Test Center for Aquatic Products of Ministry of Agriculture, East China Sea Fisheries Research Institute,
    Chinese Academy of Fishery Sciences, Shanghai 200090, China
  • Received:2010-09-25 Revised:2011-02-14 Online:2011-03-15 Published:2011-03-03
  • Contact: YANG Qiao E-mail:qyang@mail.whu.edu.cn

摘要: 选取浅玫瑰链霉菌、红色糖多孢菌及黑曲霉为发酵菌种,采用Plackett-Burman设计及正交试验,对影响微生物发酵法产大豆异黄酮苷元(染料木黄酮、黄豆苷元、黄豆黄素)的连续超声波提取工艺的主要试验因素进行系统优化。以浅玫瑰链霉菌为发酵菌种的实验结果表明:各主要试验因素对3种异黄酮苷元提取率影响程度大小为:抽提溶剂组分、超声波时间、淋洗液体积。而温度、溶剂流速两个因素与提取效率负相关。连续超声波作用时间及抽提溶剂组分为影响异黄酮苷元提取工艺的两个主要影响因素。当两因素的最优值分别为7min及70%时,总异黄酮苷元提取率为(98.7±1.23)%。通过固相萃取(SPE)可有效去除发酵提取液中杂质对HPLC定量分析的干扰,并使3种异黄酮苷元含量浓缩10倍以上,从而使HPLC方法的检测限达到10-3μg/mL,样品总分析时间小于10min,样品回收率为96.2%~103.4%,RSD值小于2.34%。上述结果表明,该方法可满足微生物发酵法产大豆异黄酮苷元的高效提取纯化及快速定量分析。

关键词: 大豆异黄酮苷元, 连续超声波提取工艺, 固相萃取, 微生物发酵

Abstract: The aim of the present study was to optimize the continuous ultrasonic extraction process for isoflavone aglycones including genistein, daidzein and glycitein produced by Streptomyces roseolus (ATCC 31047) based on Plackett-Burman design and orthogonal array design. Results indicated that the order of three positive factors affecting extraction efficiency from strong to weak was solvent composition (ethanol aqueous solution), ultrasonic treatment time and eluent volume. However, extraction temperature and eluent flow rate exhibited a negative impact on extraction efficiency. Ultrasonic treatment time of 7 min and solvent composition of 70% ethanol concentration were found optimal, and the extraction rate of total isoflavone aglycones reached up to (98.7 ± 1.23)% under the optimal conditions. The application of solid-phase extraction (SPE) could effectively reduce the interference of impurity substances during the quantitative analysis of the above three isoflavone aglycones so that the detection limit of the developed SPE-HPLC method could reach 10-3μg/mL. The detection procedure was less than 10 min and the recovery rate was between 96.2% and 103.4% with a RSD of less than 2.34%. Therefore, this method can meet the requirements for high-efficiency extraction and rapid quantitative analysis of isoflavone aglycones produced by microbial fermentation.  

Key words: isoflavone aglycones, continuous ultrasonic extraction, solid-phase extraction, microbial fermentation

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