白芷醇溶蛋白质提取工艺优化及其指纹图谱研究  

Study on Optimal Extraction Conditions and Fingerprint of Angelica dahurica Alcohol-soluble Proteins

作　　者：申金田 常雪君 全心怡 林奥 李海毅 闻崇炜[1] 汤建 Shen Jintian;Chang Xuejun;Quan Xinyi;Lin Ao;Li Haiyi;Wen Chongwei;Tang Jian(School of Pharmacy,Jiangsu University,Zhenjiang 212013,China;School of Traditional Chinese Medicine,Bozhou University,Bozhou 236800,China)

机构地区：[1]江苏大学药学院,江苏镇江212013 [2]亳州学院中药学院,安徽亳州236800

出　　处：《中国野生植物资源》2023年第8期23-28,共6页Chinese Wild Plant Resources

基　　金：安徽省高等学校自然科学研究项目(KJ2019ZD80);安徽省高校学科(专业)拔尖人才学术资助项目(GXBJZD50);江苏大学第18批大学生科研立项(Y18A161);江苏省大学生实践创新训练计划项目(202010299223Y)。

摘　　要：目的:基于响应面设计优化白芷醇溶蛋白质提取工艺,并研究其指纹图谱特征。方法:首先考察液料比、提取时间、乙醇浓度、提取温度四个单因素对白芷醇溶蛋白质提取率的影响,再通过Box-Behnken方法对影响提取率的工艺因素进行响应面优化,最后对白芷及易混淆药材的醇溶蛋白质指纹图谱进行对比分析。结果:最佳提取工艺条件为:液料比13∶1(mL/g),提取时间8.75 h,乙醇浓度42%,提取温度45℃,在此条件下提取率为0.294%。SDS-PAGE分析表明白芷醇溶蛋白质样品分子量范围介于15~95 kDa,91.4 kDa与65.6 kDa分子为高丰度蛋白质。此外,白芷与天花粉、粉葛、山药的醇溶蛋白质指纹图谱有显著差异。结论:利用响应面法优化了白芷醇溶蛋白质的提取工艺,并建立其专属性SDS-PAGE指纹图谱,为其分子鉴定及资源开发提供实验依据。Objective:To optimize the extraction technology of Angelica dahurica alcohol-soluble proteins(ADAP)and study its fingerprint.Methods:Firstly,the influence of four single factors of liquid-tomaterial ratio,extraction time,ethanol concentration and extraction temperature on the extraction yield of ADAP was investigated.Secondly,the technological factors affecting the extraction yield of ADAP was optimized by Box-Behnken method.Finally,the fingerprint of ADAP was compared and analyzed with that of other easily confused traditional Chinese medicines.Results:The optimum extraction conditions were as follow:liquid to material ratio of 13∶1(mL/g),extraction time of 8.75 h,ethanol concentration of 42%,and extraction temperature of 45℃.Under the above conditions,the extraction yield of ADAP was 0.249%.SDS-PAGE analysis showed that the molecular weights of ADAP were within 15~95 kDa.Both the 91.4 kDa and 65.6 kDa molecules were high abundance proteins.In addition,there were significant differences in the fingerprint of alcohol-soluble proteins between Angelica dahurica and Trichosan⁃thes kirilowii Maxim,Dioscorea opposita Thunb,Puueraria thonsonii Benth.Conclusion:The optimal extraction process and exclusive fingerprint of ADAP were established by means of response surface methodology and SDS-PAGE,respectively.These results will be helpful for further molecular identification and resource development of Angelica dahurica.

关 键 词：白芷药材 醇溶蛋白质 响应面法 提取工艺 指纹图谱 

分 类 号：R932[医药卫生—生药学] R282.5[医药卫生—药学]