Box-Behnken设计-响应面法优化经典名方四妙勇安汤煎煮工艺研究

Optimization of decocting process of Simiao Yong’an Decoction by Box-Behnken design-response surface methodology

目的遵循中医药传承创新发展的基本准则——“遵古原则”,考察经典名方四妙勇安汤(Simiao Yong’an Decoction,SYD)的煎煮工艺,确保原汤剂原汁原味的传统功效,并进行质量综合评价,为经典名方SYD进一步制剂开发提供参考。方法采用Box-Behnken设计-响应面法对加水量、浸泡时间、煎煮时间进行考察,通过指纹图谱结合多元统计分析进行定性半定量评价,通过含量测定分析和干膏率进行定量评价,综合以上评价方法优选煎煮工艺并验证。结果通过Box-Behnken设计-响应面法制备17批煎煮样品,建立了17批样品HPLC指纹图谱,标定了14个共有峰,利用多元统计分析对指纹图谱共有峰数据进行主成分分析(principal component analysis,PCA)与正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)并计算得分,结果显示9号工艺为最佳煎煮工艺,即煎煮2次,第1次加16倍量水,浸泡30 min,煎煮30 min;第2次加16倍量水,煎煮30 min。同时采用HPLC对17批样品进行定量分析,对绿原酸、阿魏酸等6个指标成分进行含量测定,结合干膏率结果进行综合评分,结果9号煎煮工艺评分最高。对实验结果的2种评价方法所得结果一致,通过工艺验证试验最终确定SYD最佳煎煮工艺。结论利用Box-Behnken设计-响应面法,结合“指纹图谱-多元统计分析-指标成分”多维评价的评控方法,得出的SYD传统最佳煎煮工艺稳定可行,为煎煮工艺不详的一类经典名方制剂研发提供一种研究模式。

Objective Following the basic principle of traditional Chinese medicine inheritance,innovation and development–“the principle of following the ancient”,the decocting process of the classic famous recipe Simiao Yong’an Decoction(四妙勇安汤,SYD)was investigated to ensure the original decoction’s original traditional efficacy,and the quality was comprehensively evaluated to provide reference for the further preparation development of the classic famous recipe SYD.Methods Box-Behnken design-response surface method was used to investigate the amount of water added,soaking time and cooking time.Qualitative and semi-quantitative evaluation was carried out through fingerprint analysis combined with multivariate statistical analysis,and quantitative evaluation was carried out through content determination analysis and dry paste rate,and the above evaluation methods were integrated to optimize the decocting process and verify.Results A total of 17 batches of decocting samples were prepared by Box-Behnken design-response surface method,and HPLC fingerprints of 17 batches of samples were established,and 14 common peaks were calibrated.Multivariate statistical analysis was used to perform principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)on the common peak data of fingerprint,and the scores were calculated.The results showed that No.9 was the best decocting process,that is,decocting for two times,adding 16 times of water to the first time,soaking for 30 min,decocting for 30 min;Add 16 times amount of water for the second time and cook for 30 min.At the same time,17 batches of samples were quantitatively analyzed by HPLC,and six index components such as chlorogenic acid and ferulic acid were determined.Combined with the results of dry paste rate,the score of No.9 decocting process was the highest.The results obtained by the two evaluation methods were consistent,and the best decocting technology of SYD was finally determined through the process verification test.Conclusion By using Box-Behnken design-response surface method,combined with the multidimensional evaluation and control method of“fingerprint-multivariate statistical analysis-index components”,the optimal traditional decocting process of SYD was stable and feasible,which provided a research model for the development of a class of classic famous recipes with unknown decocting process.