ASE-HPLC法同时测定壮骨伸筋胶囊中4个成分的含量

目的：建立快速溶剂萃取（ASE）-高效液相色谱法（HPLC）同时测定壮骨伸筋胶囊中葛根素、松果菊苷、柚皮苷和淫羊藿苷的含量。方法：利用正交试验优化快速溶剂萃取仪参数，得到最优提取条件，再采用HPLC法同时测定壮骨伸筋胶囊中葛根素、松果菊苷、柚皮苷和淫羊藿苷的含量。采用Phenomenex Kintex C₁₈（4.6 mm×100 mm，2.6 μm）色谱柱，流动相为乙腈-0.1%磷酸水溶液梯度洗脱，流速0.5 mL·min^-1；检测波长为290 nm；柱温为30℃。结果：采用甲醇为萃取溶剂，萃取温度为100℃、静态萃取时间为3 min以及循环次数2次的方法最优，葛根素、松果菊苷、柚皮苷和淫羊藿苷进样量分别在4.4~109.7 μg（r=0.999 8）、7.5~187.8 μg（r=0.999 7）、2.4~58.9 μg（r=0.999 8）和4.2~104.8 μg（r=0.999 9）范围内呈良好的线性关系，平均回收率在96.3%~101.3%之间，均符合含量测定的要求，测定样品10批，葛根素、松果菊苷、柚皮苷、淫羊藿苷含量范围分别在1.392~1.520、1.892~2.146、0.335~0.381、0.887~0.925 mg·g^-1之间，ASE与药典方法结果无显著差异。结论：本文建立的测定方法具有准确、快速、简便等特点，提升了壮骨伸筋胶囊的质量标准。

Objective: To establish a method of accelerated solvent extraction (ASE)-high performance liquid chromatography (HPLC)for simultaneous determination of puerarin, echinacoside, naringin and icariin in Zhuanggu Shenjin capsules. Methods: Based on orthogonal test for the optimal extraction conditions in the solvent extraction instrument for parameter extraction analysis, the HPLC method was used for determination of puerarin, echinacoside, naringin, and icariin in Zhuanggu Shenjin capsules. The chromatographic column was Phenomenex Kintex C₁₈ column (4.6 mm×100 mm, 2.6 μm), the mobile phase was acetonitrile-0.1% phosphoric acid solution with gradient elution, the flow rate was 0.5 mL·min^-1;Detection wavelength was 290 nm;column temperature was 30℃. Results: Methanol was used as extraction solvent, the extraction temperature was 100℃, the static extraction time was 3 min and the number of cycles was 2 times. It showed a good linear relationship of puerarin, echinacoside, icariin and naringin was in the range of 4.4-109.7 μg (r=0.999 8), 7.5-187.8 μg (r=0.999 7), 2.4-58.9 μg (r=0.999 8)and 4.2-104.8 μg (r=0.999 9), respectively. The measured average recovery rate was between 96.3% and 101.3%. The contents of puerarin, echinacoside, naringin, and icariin in 10 batches of samples were in the range of 1.392-1.520 mg·g^-1, 1.892-2.146 mg·g^-1, 0.335-0.381 mg·g^-1, and 0.887-0.925 mg·g^-1, respectively. The results obtained from ASE-HPLC had no significant difference with that from the method of Chinese Pharmacopoeia. Conclusion: The established method in this paper is accurate, fast and convenient, and provide the basis for improving the quality control of Zhuanggu Shenjin capsules.