基于Box-Behnken设计-响应面法与质量综合评价优化经典名方桃红四物汤煎煮工艺

Optimization of decocting process of Taohong Siwu Decoction based on BoxBehnken response surface methodology and comprehensive quality evaluation

目的遵古优化和科学评价桃红四物汤(Taohong Siwu Decoction,TSD)的煎煮工艺,为其经典名方中药复方制剂研究奠定基础。方法以指纹图谱概貌结合多指标定量为综合评价,采用Box-Behnken设计-响应面设计对加水量、浸泡时间、煎煮时间、煎煮次数进行优化。指纹图谱研究采用SunFire-C18色谱柱(150 mm×3.0 mm,3.5μm),流动相为乙腈-0.5%磷酸水溶液,体积流量为0.2 mL/min,柱温为25℃,检测波长为230 nm;建立HPLC指纹图谱,对色谱信息进行主成分分析,进一步建立UPLC多波长检测方法测定6个关键质控成分含量,计算各样品综合得分,通过响应面法进行工艺优化。结果建立了29个试验号TSD UPLC指纹图谱,选择16个共有峰,指认出地黄苷D、苦杏仁苷、羟基红花黄色素A、芍药苷、阿魏酸、藁本内酯共6个成分。主成分分析提取出4个主成分,综合得分结果显示15号工艺为最佳工艺。含量测定显示,地黄苷D、苦杏仁苷、芍药苷、阿魏酸、藁本内酯、羟基红花黄色素A 6种成分线性关系良好,综合得分结果显示,15号工艺为最佳工艺。进一步根据回归模型拟合,得到最优工艺为加水量9.90倍、浸泡时间0.58 h、煎煮时间1.19 h、煎煮次数1.60次,最终确定TSD煎煮工艺为加10倍量水,浸泡0.5 h,煎煮2次,每次1 h。结论指纹图谱质量概貌结合关键质控成分可作为TSD制备工艺的质量评控方法,得出的最佳煎煮工艺稳定可行,可用于经典名方TSD复方制剂进一步研究开发。

Objective To optimize and scientifically evaluate of the decoction process of Taohong Siwu Decoction(桃红四物汤,TSD)according to ancient law,so as to lay a foundation for the study of traditional Chinese medicine compound preparation of classical prescription.Methods Using fingerprint profiles combined with multi-index quantitative determination as a comprehensive evaluation,Box-Behnken response surface design was used to optimize water consumption,soaking time,time of decoction,and frequency of decoction.Fingerprinting was performed on the SunFire-C18 column(150 mm×3.0 mm,3.5μm)with acetonitrile-0.5%phosphoric acid aqueous solution as mobile phase,flow rate was 0.2 mL/min,column temperature was 25℃,and detection wavelength was 230 nm.The HPLC fingerprint was established,and the comprehensive score of each test sample was calculated by principal component analysis of the chromatographic peaks.Furthermore,the content of the six quality control components was detected by UPLC multiwavelength detection,each sample comprehensive score was calculated,and decoction process was optimized by response surface methodology.Results The HPLC fingerprint of 29 test samples of TSD was established.A total of 16 chromatographic peaks were selected as the common peaks of the fingerprint and six ingredients were identified including rehmannioside D,amygdalin,paeoniflorin,ferulic acid,hydroxysafflor yellow A,ligustilide with good linear relation.The result of comprehensive score showed that process number 15 was the best process.According to the regression model fitting,the optimal process was obtained as follows:adding 9.90 times of water,soaking time 0.58 h,decocting time 1.19 h and decocting times 1.60 times.Finally,the decocting process of TSD was determined as follows:adding 10 times of water,soaking for 0.5 h,decocting twice,1 h for each time.Conclusion Fingerprint quality profile combined with key quality control components can be used to evaluate the quality of the preparation process of TSD.The optimal decoction process is stable and feasible,which can be used for further research and development of the classical prescription TSD.