Box-Behnken响应面法优化橘红提取液的制备工艺

Optimization of the preparation process of Citrus grandis Tomentosa extract by Box-Behnken response surface methodology

目的:通过设定不同工艺参数制备化橘红醇提液,并测定其柚皮苷含量,明确最佳制备工艺。方法:使用回流法制备化橘红醇提液,考察参数为提取时间、乙醇含量和提取次数,使用Design Expert软件,Box-Behnken响应面法设计各实验组及相关参数,所得样品使用高效液相色谱法测定柚皮苷含量,作为考察指标;色谱条件为,Waters XBridge C18色谱柱(规格250mm*4.6mm,填料粒径5μm),流动相为乙腈(A)-0.2%甲酸水溶液(B),梯度洗脱,程序为,0-15min,20%-20%A;15-20min,20%-90%A;20-25min,90%-20%A;流速1.0ml/min,检测波长283nm,柱温25℃;共设计18组实验,得18组样品并测定其柚皮苷含量。结果:18批样品中,以提取时间50min,乙醇含量60%,提取次数2次条件所制备样品为最优,提取液中化橘红含量为0.2684-0.2710mg/ml。结论:本文所用实验设计方法能够高效、准确地得出化橘红提取液的最优制备工艺。

Objective:To prepare Citrus grandis Tomentosa alcohol extract by setting different technological parameters,and determine the content of naringin,so as to find the best preparation technology.Methods:The reflux method was used to prepare the alcohol extract of Citrus grandis Tomentosa,and the parameters were extraction time,ethanol content and extraction times.The experimental groups and related parameters were designed using the Design Expert software and Box-Behnken response surface method,and the content of naringin in the obtained samples was determined by high performance liquid chromatography as the investigation index.The chromatographic conditions were as follows:Waters XBridge C18 column(250 mm*4.6 mm,packing particle size 5μm),mobile phase consisted of acetonitrile(A)-0.2% formic acid aqueous solution(B),gradient elution,0-15 min,20%-20%A;15 to 20 min,A 20%-90%;20 to 25 min,A 90%-20%;The flow rate was 1.0 mL/min,the detection wavelength was 283 nm,and the column temperature was 25℃.A total of 18 groups of experiments were designed,18 groups of samples were obtained and the content of naringin was determined.Results:Among 18 batches of samples,the optimal conditions were extraction time 50 min,ethanol content 60% and extraction cycle 2 times.The content of orange in the extract was 0.2684-0.2710 mg/mL.Conclusion:The experimental design method used in this paper can efficiently and accurately obtain the optimal preparation process of the extract of Citrus chinensis.