Box-Behnken设计-响应面法优化醋艾叶炮制工艺及其对镇痛活性的影响

Optimization of Processing Technology of Vinegar-Processed Artemisiae Argyi Folium by Box-Behnken Design-Response Surface Method and Its Effect on Analgesic Activity

    摘要
  • 摘要:
    目的 优选醋艾叶最佳炮制工艺，并分析炮制前后指标成分含量及镇痛活性差异。
    方法 分别以异泽兰黄素和Santamarine为对照品，建立艾叶中总黄酮和总萜含量测定方法；采用HPLC-VWD建立同时测定艾叶中6-Methoxytricin、Santamarine、异泽兰黄素及Artemisolide 4种活性成分含量的方法。以总黄酮、总萜和4种活性成分含量为考察指标，以米醋用量、闷润时间、炒制温度和炒制时间为考察因素，采用Box-Behnken设计-响应面法优化醋艾叶的最佳炮制工艺。同时采用定量分析结合聚类分析(Hierarchical cluster analysis，HCA)、主成分分析(Principal component analysis，PCA)和正交偏最小二乘判别分析(Orthogonal partial least squares discriminant analysis，OPLS-DA)开展优化工艺的验证性实验，评价炮制前后指标成分的含量差异，采用福尔马林致痛小鼠模型考察醋艾叶炮制前后镇痛活性差异。
    结果 醋艾叶最佳炮制工艺参数为米醋用量16%、闷润时间50 min、炒制温度160 ℃、炒制时间12 min。验证性实验结果证实炮制前后指标成分含量存在显著差异，且醋艾叶镇痛活性显著优于艾叶。
    结论 优选出的醋艾叶炮制工艺稳定可行，可为规范醋艾叶的标准化生产提供科学依据。

     

    Abstract:
    OBJECTIVE To optimize the optimum processing technology of vinegar-processed Artemisiae Argyi Folium (AAF), and analyze the difference of index components and analgesic activity before and after processing.
    METHODS The method for the determination of total flavonoids and total terpenes in AAF was established with eupatilin and Santamarine as control substances, respectively. The HPLC-VWD method was established for the simultaneous determination of 6-Methoxytricin, Santamarine, eupatilin and Artemisolide in AAF. Box-Behnken design-response surface method was used to optimize the processing technology of vinegar-processed AAF, with the content of total flavonoids, total terpenes and four active ingredients as the index, and the amount of rice vinegar, moistening time, processing temperature and processing time as the factors. The quantitative analysis was combined with hierarchical cluster analysis (HCA), principal component analysis(PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) to evaluate the difference of index components before and after processing. The analgesic activity before and after processing was investigated using pain mice induced by formalin.
    RESULTS The optimum processing parameters of vinegar-processed AAF were 16% rice vinegar dosage, moistening time 50 min, processing temperature 160 ℃ and processing time 12 min. There were significant differences in the content of index components before and after processing. The analgesic activity of vinegar-processed AAF was significantly better than that of AAF.
    CONCLUSION The optimized processing technology is stable and feasible, which can provide scientific basis for standardizing production of vinegar-processed AAF.

     

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