用于黄芩提取物质量评价及制法影响因素考察的HPLC特征图谱的构建

Study on HPLC-characteristic chromatogram for evaluation of quality of Scutellaria Extract and influencing factors of preparations

建立并验证黄芩提取物特征图谱方法,对市售黄芩提取物质量及影响质量的制法因素进行评价。采用HPLC特征图谱方法对不同厂家、不同规格、不同批次的黄芩提取物及含黄芩提取物的制剂进行检测,以特征峰为评价指标考察制法工艺对质量的影响,并采用化学识别模式对特征峰数据进行分析。研究结果表明,建立的特征图谱方法具有良好的精密度、重复性和稳定性;31份黄芩提取物样品和4份制剂中有10个特征峰,提取物制法工艺影响黄芩提取物特征峰的峰面积差异;相似度分析结果显示:除去黄芩苷后,其中9份样品特征图谱的相似度<0.90,说明市售黄芩提取物质量具有差异性;非参数检验结果显示:5个厂家黄芩苷规格为85%的样品中4号、6号、7号特征峰相对峰面积分别具有显著性差异(P<0.05),说明黄芩提取物质量差异性主要体现在4号、6号、7号特征峰(分别为去甲汉黄芩素-7-O-葡萄糖醛酸苷、千层纸素A苷、汉黄芩苷);聚类分析根据10个特征峰将31份黄芩提取物样品和4份制剂归为5类,而主成分分析对提取物特征峰综合评价并对其质量进行排序,结果表明黄芩苷含量越高的样品,其得分值(F)越低,从统计学角度评价黄芩提取物质量应以多成分为指标,而非单一黄芩苷成分。该研究所建立的HPLC特征图谱方法准确、稳定,结合化学模式识别研究有利于黄芩提取物及含黄芩提取物的制剂的整体质量控制。

This work was carried out to establish and validate the specific characteristic chromatography of Scutellaria Extract and evaluate the quality of commercial Scutellaria Extract and optimize production process parameters. Scutellaria Extract and formulation samples collected from different manufacturers were analyzed using HPLC-characteristic chromatogram. The HPLC characteristic peaks were considered to be the index for the parameters optimization in preparations of Scutellaria Extract. The chemical pattern recognization was applied for the analysis of characteristic peak data. The established characteristic fingerprint method possessed a good precision, repeatability, and stability. Ten common characteristic peaks were determined and identified in the specific chromatogram of 31 batches of Scutellaria Extract and 4 batches of formulation. The process parameters influenced the characteristic peaks area significantly. The similarity analysis results showed that after removing baicalin peak, the similarity of the characteristic chromatography of 9 samples were less than 0.90, indicating that the quality of commercial Scutellaria Extract was inconsistent. Non-parametric test result showed that the relative peaks area of the number 4, 6, 7 characteristic peaks had significant differences(P<0.05), indicating the differences of the Scutellaria Extract quality were mainly reflected in the characteristic peaks 4, 6 and 7(norwogonin-7-O-glucuronide, oroxylin A-7-O-glucuronide, wogonoside, respectively). The cluster analysis accurately classified 31 extract samples and 4 formulation samples into five categories based on 10 characteristic peaks. The quality of Scutellaria Extracts was comprehensively evaluated and ranked by PCA. The result showed that the higher content of baicalin, the score value of quality(F value) was lower, and the quality of the Scutellaria Extract should be evaluated by multi-components instead of a single baicalin component. The optimized characteristic peaks method is stable and reliable. It is advantageous for the extract and formulations integrative quality control by combining the chemical pattern recognization.