毛冬青质量标准的改进研究

目的：完善毛冬青药材质量标准，提高其质量控制水平。方法：采用《中华人民共和国药典》2015年版四部相关方法，对毛冬青药材的水分、灰分、浸出物（水溶性和醇溶性）及有害物质含量进行测定。采用显微鉴别法对毛冬青药材的根、茎及粉末进行鉴别，并建立毛冬青药材薄层色谱及指纹图谱鉴别方法。同时以冬青素A为指标成分，建立毛冬青中冬青素A的HPLC含量测定方法。结果：毛冬青横切面和粉末的显微特征明显，薄层色谱斑点清晰，冬青素A分离较好；以冬青素A为参照峰，通过比较10批毛冬青药材的HPLC图谱中共有峰，确定了5个共有特征峰，建立了毛冬青药材的HPLC指纹图谱；冬青素A质量浓度在25~400 mg·L^-1范围内线性关系良好，线性回归方程为Y=5882X+19706（R²=0.9999，n=5），平均回收率为95.5%（RSD=3.1%，n=5）。10批毛冬青的水分为5.79%~7.43%，总灰分为1.00%~2.36%，酸不溶性灰分为0.20%~0.57%，水溶性浸出物为3.41%~10.18%，醇溶性浸出物为5.81%~15.33%，冬青素A含量为0.11%~1.56%。建议规定毛冬青的水分限度 ≤ 8.00%，总灰分限度 ≤ 2.50%，酸不溶性灰分 ≤ 0.50%，水溶性浸出物限度 ≥ 4.00%，醇溶性浸出物限度 ≥ 7.00%，以干燥品计，冬青素A的含量 ≥ 0.26%。结论：本研究完善和改进了毛冬青药材的质量控制方法。 

Objective: To improve the quality standard of Ilicis Pubescentis Radix et Caulis and improve its quality control level. Methods: Water content, total ash and extractives (water soluble and ethanol soluble)of Ilicis Pubescentis Radix et Caulis were determined according to the Pharmacopoeia of the Peoples's Republic of China (2015 edition, volume 4). Microscopic characteristics were studied based on transverse section and powder. TLC method and HPLC fingerprint chromatographic were established. The HPLC method for the determination of Ilexgenin A in Ilicis Pubescentis Radix et Caulis was developed and validated with ilexgenin A as the reference substance. Results: Microscopic characteristics of transverse section and powder of Ilicis Pubescentis Radix et Caulis were characteristic, the TLC spots were clear, and the separation of ilexgenin A was good. Five common peaks were selected as the fingerprint peaks by comparing the HPLC chromatographic of 10 batches of Ilicis Pubescentis Radix et Caulis with Ilexgenin A as the reference substances. The linear range of ilexgenin A was 25-400 mg·L^-1, the linear regression equation was Y=5 882X+19 706 (R²=0.999 9, n=5)and the average recovery was 95.5% (RSD=3.1%, n=5). The range for the water, total ash, acid insoluble ash, ethanol soluble extractives, water extractives, and ilexgenin A contents of 10 batches were 5.79%-7.43%, 1.00%-2.36%, 0.20%-0.57%, 5.81%-15.33%, 3.41%-10.18% and 0.11%-1.56% respectively. Thus, the limit of each test items for Ilicis Pubescentis Radix et Caulis were proposed as follows:water ≤ 8.00%, total ash ≤ 2.50%, acid insoluble ash ≤ 0.50%, water extractives ≥ 4.00%, ethanol soluble extractive ≥ 7.00%, the content of ilexgenin A calculated on dried sample ≥ 0.26%. Conclusion: The established method improves the quality control of Ilicis Pubescentis Radix et Caulis. 

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