[关键词]
[摘要]
目的 建立HPLC指纹图谱检测方法,探寻青龙白虎汤冻干粉制备过程工艺和量质传递规律,并结合化学计量学构建其质量控制体系。方法 制备15批次青龙白虎汤冻干粉,采用HPLC法建立指纹图谱,色谱结果导入《中药色谱指纹图谱相似度评价软件》(2012版)并计算各部分相似度。测定15批样品量质传递过程中没食子酸、原儿茶酸、绿原酸、表儿茶素和鞣花酸的含量,计算转移率和出膏率。结合化学计量学方法进行分析,以挖掘不同产地样品间对质量控制具有显著贡献的主要成分。结果 15批样品指纹图谱相似度均大于0.9,满足规定要求,并标定冻干粉指纹图谱24个共有峰,对其中6种成分进行了指认,分别为没食子酸(1号峰)、原儿茶酸(4号峰)、绿原酸(8号峰)、表儿茶素(14号峰)、东莨菪内酯(19号峰)、鞣花酸(23号峰)。15批冻干粉表儿茶素、没食子酸、鞣花酸、绿原酸、原儿茶酸的质量分数分别为0.724%~1.301%、2.184%~2.840%、0.607%~0.760%、0.061%~0.141%、0.017%~0.079%,转移率分别为78.60%~89.38%、76.98%~89.88%、76.00%~89.78%、76.90%~90.49%、80.02%~90.25%,出膏率为12.87%~15.11%,均未出现离散数据,表明煎煮、浓缩和冻干过程有效成分转移率较稳定。通过化学计量学分析,找到10个对模型贡献较大的成分,其中包括指认的峰14 (VIP=2.812)、峰1(VIP=2.804)、峰23(VIP=2.715)、峰8(VIP=1.053)和峰4(VIP=0.887),可进一步深化青龙白虎汤质量控制研究。结论 通过HPLC指纹图谱结合多指标成分含量测定,首次建立了药食同源名方青龙白虎汤的质量控制方法,此方法快速简单可行,重复性、稳定性良好,能同时适用于饮片、煎煮液、浓缩液和冻干粉量质传递规律的相关性考察;进一步通过化学计量学为青龙白虎汤的质量控制研究提供了重要参考。
[Key word]
[Abstract]
Objective To explore the preparation process of lyophilized powder of Qinglong Baihu Decoction (青龙白虎汤, QBD) and the law of quantity-quality-correlation by establishing an HPLC fingerprint detection method, and build its quality control system combined with chemometrics. Methods Fifteen batches of QBD lyophilized powder were prepared, the fingerprints were established by HPLC method, then imported the chromatographic results into TCM Chromatographic Fingerprint Similarity Evaluation Software (2012) and calculated the similarity of each part. The content of gallic acid, protocatechuic acid, chlorogenic acid, epicatechin, and ellagic acid were detected during the quantity-quality-correlation of 15 batches of samples, and the transfer rate and extractum rate were calculated. Combined with the chemometrics methods, the main components of samples from different producing areas which had significant contribution to the quality control were analyzed. Results The similarity of the fingerprints of 15 batches of samples was > 0.9, which met the specified requirements, and the 24 common peaks of the fingerprints of the lyophilized powder were calibrated, and six of them were identified, they were gallic acid (peak 1), protocatechuic acid (peak 4), chlorogenic acid (peak 8), epicatechin (peak 14), scopoletin (peak 19), and ellagic acid (peak 23). The content range of these components in 15 batches of lyophilized powder were 0.724%—1.301% epicatechin, 2.184%—2.840% gallic acid, 0.607%—0.760% ellagic acid, 0.061%—0.141% chlorogenic acid, 0.017%—0.079% protocatechuic acid, the transfer rates of epicatechin, gallic acid, ellagic acid, chlorogenic acid and protocatechuic acid were 78.60%—89.38%, 76.98%—89.88%, 76.00%—89.78%, 76.90%—90.49%, 80.02%—90.25% respectively, the extractum rate was 12.87%—15.11%, and without discrete data. It showed that during the process of decoction, concentration and lyophilized powder, the transfer rates of effective components were stable. Through chemometrics analysis, it was found that 10 component scontributed significantly to the model, including the designated peak 14 (VIP = 2.812), peak 1 (VIP = 2.804), peak 23 (VIP = 2.715), peak 8 (VIP = 1.053) and peak 4 (VIP = 0.887), which can further deepen the quality control research of QBD. Conclusion The quality control method of the famous prescription with medicine and food homology of QBD was established for the first time through the HPLC fingerprint combined with the content determination of multiple index components. This method is rapid, simple, feasible, reproducible, stable and it can be applied to the correlation investigation of the quantity-quality-correlation law of decoction pieces, decoction, concentrate and lyophilized powder at the same time; Further through the chemometrics, which provided an important reference for the quality control research of QBD.
[中图分类号]
R283.6
[基金项目]
四川省中医药管理局中医药开发专项(2018KF001);四川省科技厅国际合作项目(2018HH0122);成都市科技局国际科技合作项目(2017-GH02-00054-HZ);成都中医药大学科技转化项目(CGPY1605)