首页|基于凝血活性和星点设计-效应面法的黄槐复方提取工艺研究

基于凝血活性和星点设计-效应面法的黄槐复方提取工艺研究

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运用纤维蛋白原平板法,以凝血酶为参照,建立体外凝血活性测定的方法,以体外凝血活性为指标,筛选黄槐复方提取路线,并在单因素试验基础上,以黄芩苷、芦丁提取转移率和浸膏得率的综合评分值为因变量,乙醇浓度、溶剂用量、提取时间为影响因素,利用星点设计3因素5水平试验,进行二次多项式拟合,通过效应面法优选黄槐复方提取工艺,并测定其凝血活性.结果表明凝血酶浓度在0.4~16 U·mL-1与沉淀圈面积呈良好线性关系,r=0.997 5;平均加样回收率为103.8%,RSD4.7%;黄槐合提浸膏的沉淀圈面积38.81 mm2,比分提浸膏沉淀圈面积大;黄槐浸膏的日用量的活性84.28 U.黄槐复方的最佳提取工艺为加6倍量40%乙醇,提取3次,每次2h,综合评分值预测值为94.26,验证值为88.34,与预测值偏差为6.28%,最佳提取工艺制得的中间体凝血活性较好.所建立的方法可以简便、快速、准确地测定黄槐复方的凝血活性,可用于后续工艺的筛选以及质量控制.
Optimization of extraction technology from compound Huanghuai based on coagulation activity and central composite design-response surface methodology
To establish a method for the determination of coagulation activity in vitro by using fibrinogen plate method.The extraction route of compound Huanghuai was optimized by selecting thrombin as the reference substance.The comprehensive score of extract yield and the extraction transfer rate of baicalin and rutin was set as the dependent variable,with alcohol concentration,solvent volume and extraction time as the influence factors in central composite design for quadratic fitting,and the extraction process of compound Huanghuai was optimized by using response surface methodology.The results of thrombin concentration and precipitation zone area showed a good linear relationship in 0.4-16 U · mL-1,r =0.997 5.The average recovery rate was 103.8% and the RSD was 4.7 %.The circle of precipitation area of compound Huanghuai combined extract was 38.81 mm2,which was bigger than that of fractionated extract.The activity on the daily amount of compound Huanghuai extract was 84.28 U;and the optimun extraction technology was as follows:alcohol concentration 40%,extracted 3 times,the liquid-solid ratio 6:1,and extraction time 2 h.The predicted value of comprehensive score was 94.26 and the measured value was 88.34,respectively,with a relative deviation of 6.28%.The coagulation activity of the intermediate obtained by optimal extraction process was better.So the method established in this paper was simple,fast and accurate for determination of coagulation activity of conpound Huanhuai,which can be also used for the screening of follow-up process and quality control.

compound Huanghuaicoagulation activityfibrinogen plate methodcentral composite design-response surface methodology

李鹏程、刘涛、伍月、张倩、付春梅、刘翠

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成都大学四川抗菌素工业研究所,四川成都610052

成都大学药学与生物工程学院,四川成都610106

黄槐复方 凝血活性 纤维蛋白原平板法 星点设计-效应面法

四川省千人计划项目

2013332

2017

中国中药杂志
中国药学会

中国中药杂志

CSTPCDCSCD北大核心
影响因子:1.718
ISSN:1001-5302
年,卷(期):2017.42(2)
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