基于指纹图谱、含量测定和化学模式识别的五子衍宗丸质量评价研究

Study on quality evaluation of Wuziyanzong pills based on HPLC fingerprint, assay and chemical pattern recognition

目的:建立五子衍宗丸指纹图谱并进行化学模式识别分析,测定五子衍宗丸中金丝桃苷、毛蕊花糖苷、山柰酚、五味子醇甲、五味子甲素和五味子乙素的含量,为该制剂的质量控制提供参考。方法:采用高效液相色谱法,使用Agilent SB-C_(18)色谱柱(250 mm×4.6 mm, 5μm),以乙腈-0.05%磷酸水溶液为流动相,梯度洗脱,流速1.0 mL·min^(-1),检测波长254 nm,柱温30℃,进样量20μL。以五味子醇甲为参照,使用《中药色谱指纹图谱相似度评价系统软件(2012A版)》建立30批五子衍宗丸的指纹图谱,确定共有峰并进行相似度评价,通过与对照品溶液比对指认色谱峰。采用Simca-P 14.1软件对共有峰进行评价。采用上述HPLC方法测定30批样品中指认化学成分的含量。结果:建立五子衍宗丸指纹图谱,共标定17个共有峰,相似度不低于0.90;共指认化学成分6个,分别为金丝桃苷、毛蕊花糖苷、山柰酚、五味子醇甲、五味子甲素、五味子乙素。经化学模式分析,30批样品聚成3类,筛选出8个差异性标志物。30批样品中上述6个成分的含量分别为1.158~3.695、0.089~1.965、1.158~3.695、0.089~1.965、1.158~3.695、0.089~1.965 mg·g^(-1)。结论:所建立的五子衍宗丸HPLC分析方法简便、稳定、可行,同时指纹图谱结合化学模式识别分析可为五子衍宗丸质量控制标准的制定提供参考。

Objective:To establish fingerprint of Wuziyanzong pills, analyze its chemical pattern recogniton, and determine the contents of hyperoside, verbascoside, kaempferol, schisandrin, deoxyschizandrin and γ-schisandrin in Wuziyanzong pills, so as to provide reference for the quality control of the preparation. Methods: HPLC method was adopted. The determination was performed on an Agilent SB-C_(18)column(250 mm×4.6 mm, 5 μm) with acetonitrile-0.05% phosphoric acid water as the mobile phase(gradient elution) at a flow rate of 1.0 mL·min^(-1). The detection wavelength was set at 254 nm, and the column temperature was 30 ℃. The sample volume was 20 μL. With schisandrin as the reference, the HPLC fingerprints of 30 batches of Wuziyanzong pills were established with Similarty Evalutation System for TCM Chromatographic Fingerprint(2012A edition);common peaks were determined and the similarity was evaluated. The chromatographic peaks were identified by comparing with the reference. Simca-P 14.1 software was used to evaluate the common peaks. The contents of the identified components in 30 batches of samples was determined by the abvoe HPLC method. Results: The fingerprint of Wuziyanzong pills was established, and 17 common peaks were calibrated, and the similarity was no less than 0.90. There were 6 chemical components identified, which were hyperoside, verbascoside, kaempferol, schisandrin, deoxyschizandrin and γ-schisandrin. 30 batches of samples were classified into three groups and eight difference markers were selected by chemical pattern recognition. The contents of above 6 components in 30 samples were 1.158-3.695 mg·g^(-1), 0.089^(-1).965 mg·g^(-1), 1.158-3.695 mg·g^(-1), 0.089^(-1).965 mg·g^(-1), 1.158-3.695 mg·g^(-1), 0.089^(-1).965 mg·g^(-1), respectively. Coclusion: The established HPLC analysis method of Wuziyanzong pills is sample, stable and feasible, and the fingerprint combined with chemical pattern recognition can provide reference for the establishment of the quality control standard of the Wuziyanzong pills.