青白通痹方药的半仿生-生物酶法提取工艺优选

SBEE Optimization of Qingbai Tongbi Prescriptions

目的:优选青白通痹方药用"半仿生-生物酶法"提取(semi-bionic enzyme extraction,SBEE)的工艺条件。方法:先选择适宜的生物酶对青白通痹方药预处理,再按照"半仿生法"(semi-bionic extraction,SBE)的操作,于第一次提取和第三次提取的溶剂中加入适宜的酶,在相应的温度下进行提取,采用L18(37)正交表,布点正交实验,以青藤碱、芍药苷、甘草次酸、分子量≤1000Da提取物、总生物碱的量、干浸膏量进行多指标综合评判,优选出三次提取溶剂的pH值,酶的种类和酶的用量,提取的温度,预处理及提取时间。结果:青白通痹方药SBEE最佳提取的工艺条件:浸提溶剂的pH值为2.0、7.5、8.5,酶Ⅰ+酶Ⅱ组合为2.0+0%,酶Ⅲ+酶Ⅳ组合为0.5%+1.0%,浸提温度为(61±1)℃,预处理和浸提时间(组序)为组2。结论:从大生产实际出发考虑,确定将青白通痹方药先用2%纤维素酶(以方药重量计)预处理24 h;提取3次,浸提用水的pH值依次为:2.0、7.5、8.5,并在第一次提取中加入0.5%胃蛋白酶,第三次加入1%胰酶;提取温度为:(61±1)℃;提取时间依次为:1.5、1、1 h。

Objective:To optimize SBEE of Qingbai Tongbi prescription.Methods:Suitable enzyme was selected to pretreat Qingbai Tongbi prescription,according to the operation of semi-bionic extraction(SBE),adding the suitable enzyme to the solvent in the first extraction and the third extraction,and they were extracted under the corresponding temperature,by using L18(37)orthogonal table,the layout of orthogonal design,sinomenine,paeoni-florin,glycyrrhetinic acid,the extract of molecular weight≤1000Da,total alkaloids amounts and dry extract were used to perform multi-index comprehensive evaluation,to optimize pH value of the solvent in the extraction of three times,the types and doses of enzyme,the extraction temperature,precondition and extraction time.Results:The best extraction conditions of SBEE for Qingbai Tongbi prescription were:pH values of extraction solvents were 2.0,7.5 and 8.5,the combination of enzymeⅠ+enzymeⅡwere 2.0%+0%,the combination of enzymeⅢ+enzymeⅣwere 0.5%+1.0%,extraction temperature was(61±1)℃,preconditioning and extraction time was group No.2.Conclusion:Considering the reality of mass production,it is confirmed that preconditioning the prescription with 2%cellulase(calculated with the weight of the prescription)24 hours;extracting three times,pH values of extracting water are 2.0,7.5 and 8.5,adding 0.5%pepsin in the first extraction,1%pancreatin in the third extraction;extraction temperature(61±1)℃,extraction time are 1.5,1 and 1 hour.