多指标一测多评-响应曲面法优选蜜炙甘草的最佳炮制工艺

Optimization of best processing technology of Glycyrrhizae Radix et Rhizoma Praeparata cum Melle by multi-index QAMS-response surface methodology

目的基于响应曲面法,应用HPLC法定量测定,从多指标一测多评、综合评价的角度建立蜜炙甘草的最佳炮制工艺。方法采用HPLC定量分析,以甘草苷、甘草素、甘草查耳酮A和甘草次酸的含量作为考察指标;采用响应曲面设计法考察甘草加蜜量、浸泡焖润时间、炒制温度、炒制时间对蜜炙甘草炮制工艺的影响,优选出蜜炙甘草最佳炮制工艺。结果色谱柱为Diamonsil C18(2)(200 mm×4.6 mm,5μm);流动相为乙腈-0.1%磷酸水溶液,梯度洗脱:0~20 min,12%~32%乙腈;20~45 min,32%~70%乙腈;45~75 min,70%~97%乙腈;检测波长260 nm,柱温20℃,体积流量1 mL/min;以甘草苷为内标物,测定并计算得到甘草素、甘草查耳酮A、甘草次酸与其的相对校正因子分别为0.56、0.64、1.42;优选的蜜炙甘草最佳炮制工艺为加蜜量1/4,浸泡焖润时间15 min,炒制锅底温度为160℃,炒制时间13 min。结论该实验含量测定方法学考察结果符合要求,采用响应曲面法优选的蜜炙甘草的最佳炮制方案是可行的,为制定蜜炙甘草的质量标准和现代研究提供了科学依据。

Objective Based on response surface methodology, HPLC was applied to quantitatively determine the optimal processing technology of Glycyrrhizae Radix et Rhizoma Praeparata cum Melle(GRRPM) from the perspective of multi-index and comprehensive evaluation. Methods HPLC was used for quantitative analysis, and the content of liquiritin, liquiritigenin, licochalcone A and glycyrrhetinic acid was used as inspection indexes. Response surface methodology was used to investigate the effects of the adding amount of honey, steaming and soaking time, frying temperature and frying time on the processing technology of GRRPM, and to optimize the optimal processing technology of GRRPM. Results The chromatographic column was Diamonsil C18(2)(4.6 mm × 200 mm, 5 μm);mobile phase was acetonitrile-0.1% phosphate aqueous solution, gradient eluting: 0—20 min, 12%—32% acetonitrile;20—45 min, 32%—70% acetonitrile;45—75 min, 70%—97% acetonitrile, with detection wavelength of 260 nm, column temperature of 20 ℃, and flow rate of 1 mL/min;Using liquiritin as internal standard, the relative correction factors of glycyrrhizin, licochalcone A, glycyrrhizinic acid and their relative correction factors were determined and calculated to be 0.56, 0.64 and 1.42, respectively. The optimum processing process of GRRPM was as follows: the amount of honey was 1/4, the soaking time was 15 min, frying pan bottom temperature was 160 ℃, and frying time was 13 min. Conclusion The results of systematic adaptability investigation of the experimental content determination method meet the requirements. The best processing scheme of GRRPM optimized by response surface methodology is feasible and provides scientific basis for formulating quality standards and modern research of GRRPM.