目的 基于响应曲面技术，应用HPLC法定量测定，从多指标、综合评价的角度建立酒炖熟地黄的最佳炮制工艺。方法 采用HPLC定量分析，以地黄苷D、益母草苷、5-羟甲基糠醛（5-HMF）和毛蕊花糖苷为评价指标，采用响应曲面设计法考察地黄浸润时间、加黄酒量、炖制时间、干燥温度、干燥时间对酒炖熟地黄炮制工艺的影响，寻找出最佳炮制工艺。结果 色谱柱为Tnature-C₁₈柱（250 mm×4.6 mm，5 μm），体积流量为1 mL/min，柱温为25℃。地黄苷D和益母草苷定量测定：流动相为乙腈-0.1%磷酸水溶液（4：96），检测波长为203 nm；5-HMF和毛蕊花糖苷定量测定：流动相为乙腈-0.1%磷酸水溶液，梯度洗脱：0~15 min，11%乙腈；15~15.1 min，11%~16%乙腈；15.1~50 min，16%乙腈；检测波长为334 nm。酒炖熟地黄的最佳工艺为100 kg生地加入黄酒60 kg，浸润12 h，密闭炖制38 h，在76℃干燥33 h。结论 所建立的HPLC法稳定可行，适合所选成分的测定。采用响应面法建立的模型相对准确，可以较好地预测4种成分含量；优选的酒炖熟地黄炮制工艺方法可行，为制定熟地黄的质量标准和现代研究提供了科学依据。

Objective Based on response surface methodology, the best processing technique for Rehmanniae Radix Praeparata was established in the aspects of multiple evaluations and comprehensive evaluations by the quantitative determination of HPLC. Methods The content of rehmannioside D, ajugol, 5-hydroxymethyl-2-furaldehyde (5-HMF) and verbascoside was used as the evaluation index in the quantitative analysis of HPLC. The response surface methodology was used to investigate the ingestion time of Rehmanniae Radix Praeparata, the adding amount of yellow wine, the stewing time, the drying temperature and the drying time with aim to find the best processing technology. Results The column was a Tnature-C₁₈ column (250 mm×4.6 mm, 5 μm) with volume flow of 1 mL/min and column temperature of 25℃. The elution conditions for the quantitative determination of rehmannioside D and ajugol were as follow:The mobile phase was acetonitrile-0.1% phosphoric acid water (4:96), and the detection wavelength was 203 nm; That for 5-HMF and verbascoside was as follow:The mobile phase was acetonitrile-0.1% phosphoric acid water (0-15 min, acetonitrile content was 11%; 15-15.1 min, acetonitrile content was 11%-16%; 15.1-50 min, acetonitrile content was 16%) and the detection wavelength was 334 nm. The best technology for Rehmanniae Radix Praeparata with yellow wine stewing was:adding 100 kg of yellow wine 60 kg for infiltrating 12 h, sealing and stewing for 38 h, drying at 76℃ for 33 h. Conclusion The established HPLC method was stable and feasible, suitable for the determination of the selected components. The model established by the response surface method was relatively accurate and can be used to predict the content of the four components well. The optimized process with yellow wine stewing method was feasible, which provided a scientific basis for the formulation of quality standards and modern research of Rehmanniae Radix Praeparata.

R283.1

国家自然科学基金面上项目（81673601）