半仿生酶法提取柿叶中总黄酮的工艺筛选及优化
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摘要
目的:筛选提取柿叶中黄酮的方法及最优提取工艺。方法:以浸膏质量、黄酮含量为指标,比较乙醇回流法、酶法(纤维素酶、β-葡聚糖酶及二者等量混合组成的复合酶)、半仿生法和半仿生酶法(同前3种酶)提取柿叶中总黄酮的效果。以黄酮含量为指标,以料液比、回流温度和回流时间为因素,设计正交试验优化半仿生酶法提取柿叶中黄酮的工艺条件,并进行验证试验。结果:在4种提取方法中,以半仿生酶法得到的浸膏质量和黄酮含量最高,并以复合酶效果最好;优化的半仿生酶法提取工艺条件为料液比1∶14、回流温度50℃、回流时间2.0 h;验证试验中3次黄酮提取率分别为5.9%、5.8%、5.9%(RSD=0.98%,n=3)。结论:优化后的半仿生酶法提取柿叶中黄酮,提取率高且工艺稳定。
OBJECTIVE:To screen the method for extracting total flavonoids in persimmon leaves and optimize extraction technology. METHODS:Using extract quality and flavonoids content as indexes,the effects of extracting total flavonoids in persimmon leaves by ethanol refluxing method,enzyme method(cellulase,β-glucanase and complex enzyme mixed by equal amounts of both),semi-bionic method and semi-bionic-enzyme method(the same enzymes)were compared. Using flavonoids content as index,solid-liquid radio,reflux temperature,reflux time as factors,orthogonal test was designed to optimize the extraction technology conditions of flavonoids in persimmon leaves by semi-bionic-enzyme method,and the verification test was conducted. RESULTS:The extract quantity and flavonoids content by semi-bionic-enzyme method was the highest among the 4 extraction methods,and the complex enzyme was the most suitable;the optimized extracting condition of semi-bionic-enzyme method were as follows as solid-liquid radio of 1∶14,reflux temperature of 50 ℃,reflux time of 2.0 h;extraction rates of flavonoids in 3 verification tests were 5.9%,5.8%,5.9%(RSD=0.98%,n=3). CONCLUSIONS:The optimized semi-bionic-enzyme method is efficient and stable in extracting flavonoids in persimmon leaves.
OBJECTIVE:To screen the method for extracting total flavonoids in persimmon leaves and optimize extraction technology. METHODS:Using extract quality and flavonoids content as indexes,the effects of extracting total flavonoids in persimmon leaves by ethanol refluxing method,enzyme method(cellulase,β-glucanase and complex enzyme mixed by equal amounts of both),semi-bionic method and semi-bionic-enzyme method(the same enzymes)were compared. Using flavonoids content as index,solid-liquid radio,reflux temperature,reflux time as factors,orthogonal test was designed to optimize the extraction technology conditions of flavonoids in persimmon leaves by semi-bionic-enzyme method,and the verification test was conducted. RESULTS:The extract quantity and flavonoids content by semi-bionic-enzyme method was the highest among the 4 extraction methods,and the complex enzyme was the most suitable;the optimized extracting condition of semi-bionic-enzyme method were as follows as solid-liquid radio of 1∶14,reflux temperature of 50 ℃,reflux time of 2.0 h;extraction rates of flavonoids in 3 verification tests were 5.9%,5.8%,5.9%(RSD=0.98%,n=3). CONCLUSIONS:The optimized semi-bionic-enzyme method is efficient and stable in extracting flavonoids in persimmon leaves.
引文
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[3]林娇芬,林河通,谢联辉,等.柿叶的化学成分、药理作用、临床应用及开发利用[J].食品与发酵工业,2005,31(7):90-96.
[4]郭玫,董晓萍,徐文萍.柿叶的研究概况[J].甘肃中医学院学报,2000,17(S1):78-82.
[5]马新博,宫汝飞.柿叶黄酮药理作用研究进展[J].中医药学报,2012,40(4):130-132.
[6]徐浩锋,杨华俊,俞岚.中药有效成分现代提取技术研究进展[J].海峡药学,2012,24(1):13-18.
[7]薛璇玑,郭增军,戴柳江,等.半仿生酶法提取拐枣七总生物碱工艺研究[J].现代中药研究与实践,2014,28(3):48-51.
[8]张卉,赵婷婷,戴柳江,等.半仿生酶法提取甘草中甘草酸的工艺研究[J].中国现代应用药学,2013,30(9):969-972.
[9]孙福东,孙秀梅,张兆旺,等.半仿生酶法提取半夏白术天麻汤中天麻素的含量测定[J].中华中医药学刊,2008,26(8):1823-1825.
[10]赵晓莉.柿叶黄酮类化合物提取分离研究[D].西安:西北工业大学,2005.
[11]贾娜,李晓崴,宋立,等.响应面法优化柿叶总黄酮的超声波辅助提取条件[J].食品与发酵科技,2013,49(6):1-6.
[12]赵文红,赵翾,白卫东,等.柿叶黄酮化合物提取工艺研究[J].陕西科技大学学报,2007,25(5):54-58.
[13]李艳提,赵金凤,张卫明,等.罗布麻茶总黄酮含量测定方法研究[J].食品科技,2010,35(6):274-278.
[14]杨帆,张轩,丁红.柿叶中总黄酮的鉴别与含量测定[J].中国医药指南,2013,11(8):435-436.
[15]彭凯丽,李新民,李元林,等.正交试验优选复方桑菊口服液中药材的提取工艺[J].中国药房,2014,25(3):243-245.
[16]郭增军,薛璇玑,赵雅韵,等.一种半仿生酶法提取柿叶总黄酮的方法和应用:中国,CN105012372A[P].2015-11-04.