甘草对照提取物的制备及质量研究

目的：验证甘草对照提取物制备工艺的可行性及稳定性，研究用于其质量控制的方法。方法：采用水煎提取，大孔树脂富集，喷雾干燥工艺制备甘草对照提取物；采用正反相薄层色谱及特征图谱对提取物进行定性研究；采用HPLC法对提取物中芹糖甘草苷、甘草苷、芹糖异甘草苷和甘草酸进行含量测定：使用Luna C₁₈色谱柱（250 mm×4.6 mm，5 μm），流动相为乙腈（A）-0.05%磷酸溶液（B），梯度洗脱（0~8 min，0 → 19% A； 8~18 min，19% A → 35% A； 18~33 min，35% A； 33~38 min，35% A → 55% A），流速1.0mL·min^-1，检测波长为360 nm和237 nm。结果：制备得到3批各约500 g甘草对照提取物，建立的方法薄层色谱斑点清晰，特征图谱有芹糖甘草苷、甘草苷、芹糖异甘草苷、甘草素和甘草酸5个共有峰，进行了含量测定方法验证，并测定3批对照提取物中芹糖甘草苷、甘草苷、芹糖异甘草苷和甘草酸含量分别为3.347%~7.324%、4.930%~8.587%、0.942%~2.039%和14.62%~19.93%。结论：研究制订的甘草对照提取物工艺稳定可行，建立的方法可用于甘草对照提取物的质量控制，为后续应用研究奠定了良好的基础。

Objective: To verify the feasibility and stability of the preparation process of licorice reference extract, and to establish the method for its quality control. Methods: The reference extract was obtained by being extraded with water, enriched with macroporous resin and sprayed to dry. The extract was identified by normal and reversed phase TLC method as well as HPLC characteristic chromatogram. Besides, content of liquiritin apioside, liquiritin, isoliquiritin apioside and glycyrrhizic acid was determined by HPLC method. The analysis was performed on a Luna C₁₈ column(250 mm×4.6 mm, 5 μm) using acetonitrile(A) and 0.05% phosphate acid(B) as the mobile phase with gradient elution(0-8 min, 0 → 19%A; 8-18 min, 19%A → 35%A; 18-33 min, 35%A; 33-38 min, 35%A → 55%A). The flow rate was 1.0 mL·min^-1 and the detection wavelengths were 360 nm and 237 nm. Results: Three batches of licorice reference extract(about 500 g per batch) were obtained. The TLC method gave clear spots and the HPLC characteristic chromatogram showed 5 common peaks. The quantitative method was validated and the content ranges were 3.347%-7.324% for liquiritin apioside, 4.930%-8.587% for liquiritin, 0.942%-2.039% for isoliquiritin apioside and 14.62%-19.93% for glycyrrhizic acid, respectively. Conclusion: The preparation process of licorice reference extract is stable and feasible. The proposed methods were suitable for quality control. This paper provided a good basis for the following application of the reference extract.

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