基于熵权法结合Box-Behnken响应面法优化桂枝芍药知母颗粒复方提取工艺

Optimization of extraction process of Compound Guizhi Shaoyao Zhimu Granules by combination of response surface methodology with entropy weight method

目的优选桂枝芍药知母颗粒的提取参数。方法采用HPLC法和Elisa法,以桂皮醛、芍药苷、芒果苷、甘草酸提取率,出膏率及炎症因子白细胞介素-6(IL-6)水平为评价指标,采用熵权法确定各指标权重系数,结合Box-Behnken响应面法结果优化桂枝芍药知母颗粒提取工艺参数。结果桂皮醛、芍药苷、芒果苷、甘草酸分别在3.27~104.64、8.16~261.03、1.39~89.27、1.00~33.00μg/mL内呈良好的线性关系,r分别为0.9999、1.0000、0.9999、0.9999;桂枝芍药知母提取液具有抑制类风湿关节炎滑膜成纤维细胞MH7A细胞(由TNF-α造模)增殖的作用,半数抑制浓度(IC50)为1.02 mg/mL,且具有抗炎活性,采用信息熵赋权法确定各指标的客观权重,按照其确定的权重系数进行综合评价,且考虑实际情况,确定桂枝芍药知母颗粒的最佳提取工艺条件为加16倍量水,提取3次,每次1 h。验证试验结果显示预测值与实测值相比,RSD小于3%。结论优选后的工艺稳定可行,重复性好,可用于大批量生产。

Objective To optimize the extraction process parameters of Guizhi Shaoyao Zhimu Granules(GSZG).Methods Using high performance liquid chromatography and Elisa,the extraction rate of trans-cinnamaldehyde,paeoniflorin,mangiferin,glycyrrhizic acid,the dry extract yield of extracted herbs and inflammatory factor IL-6 were comprehensively evaluated.The information entropy weighting method was used to determine the objective weight of each index,and response surface methodology was adopted to optimize the extraction process parameters of GSZG.Results The method had good linear relationship within the range of 3.27—104.64μg/mL for trans-cinnamaldehyde(r=0.9999),8.16—261.03μg/mL for paeoniflorin(r=1.0000),1.39—89.27μg/mL for mangiferin(r=0.9999),and 1.00—33.00μg/mL for glycyrrhizic acid ammonium salt(r=0.9999).Guizhi-Shaoyao-Zhimu extraction had the effect of inhibiting the proliferation of MH7 A cells(molded by TNF-α)with IC50 of 1.02 mg/mL,and had anti-inflammatory activity.Entropy method was more scientific,reasonable and stable.According to the comprehensive scoring results and practical situation,it was determined that the best extraction process of the preparation was to add 16 times the amount of water,and decocted three times for 1 h each time.The RSD of the predicted value was less than 3%compared with the measured value.Conclusion The preferred process has high extraction rate,with good stability and repeatability,which is suitable for the mass production of GSZG.