基于HPLC特征图谱及化学计量学的苗药云实感冒合剂质量评价研究

Quality Evaluation of Yunshi Ganmao Mixture Based on HPLC Characteristic Chromatogram and Chemometrics

目的:建立特征性图谱和多指标定量与化学计量学相结合的云实感冒合剂质量评价方法。方法:采用WondasilTM C18-superb色谱柱(250 mm×4.6 mm,5μm),以甲醇-0.1%磷酸水溶液为流动相,梯度洗脱,检测波长为254 nm,柱温为30℃,进样量为10μl,流速为1.0 ml·min^(-1),对没食子酸、原苏木素B、鞣花酸3种成分进行含量测定和特征图谱研究,并结合药材的图谱进行化学成分归属,结合化学计量学方法对云实感冒合剂的质量进行评价研究。结果:含量测定表明没食子酸0.3270~1.3640 mg·ml^(-1)、原苏木素B 0.1640~0.6200 mg·ml^(-1)、鞣花酸0.0160~0.0810 mg·ml^(-1)。17批云实感冒合剂的HPLC特征图谱共有模式确定了25个共有峰,其中没食子酸和鞣花酸来自于云实皮和蓝布正,原苏木素B来自于云实皮,供试品分别与云实皮、蓝布正、马鞭草、生姜药材呈现9,7,11,3个相对应的色谱峰,并且17批云实感冒合剂的特征图谱相似度大于0.97,聚类分析及主成分分析均将17批云实感冒合剂聚为3类。结论:本文建立了云实感冒合剂HPLC特征性图谱,灵敏度高,重复性好,简单方便,易操作,通过对比云实感冒合剂待测样品与云实感冒合剂HPLC特征图谱共有峰的相似度,并结合化学计量学方法,可对云实感冒合剂的质量进行整体描述和评价,并确保产品质量的均一性。

Objective:To establish a quality evaluation method for Yunshi Ganmao mixture based on HPLC characteristic chromatogram and chemometrics.Methods:The separation was performed on a WondasilTM C18-superb column with mobile phase consisting of methanol and 0.5%phosphate solution with gradient elution.The UV detection wavelength was set at 254 nm,the column temperature was 30℃,the injection volume was 10μl and the flow rate was 1.0 ml·min^(-1).The content determination and characterization of gallic acid,hematoxylin B and ellagic acid were studied and the quality of Yunshi Ganmao mixture was evaluated by stoichiometry.Results:The content ranges of the three components were as follows:0.3270-1.3640 mg·ml^(-1) for gallic acid,0.1640-0.6200 mg·ml^(-1) for hematoxylin B and 0.0160-0.0810 mg·ml^(-1) for ellagic acid.The HPLC characteristic chromatogram of Yunshi Ganmao mixture was established.Twenty-five common peaks were identified.Gallic acid and ellagic acid were derived from Caesalpinia decapetala(Roth)Alston and Geum japonicum Thunb.var.chinense Bolle.Hematoxylin B was derived from Caesalpinia decapetala(Roth)Alston.The samples showed nine,seven,eleven and three chromatographic peaks corresponding to Caesalpinia decapetala(Roth)Alston,Geum japonicum Thunb.var.chinense Bolle,Verbena officinalis L.and Zingiber officinale Rose.The similarity of characteristic chromatograms of 17 batches of Yunshi Ganmao mixture was more than 0.97.Cluster analysis and principal component analysis all classified the 17 batches of Yunshi Ganmao mixture into 3 categories.Conclusion:The HPLC characteristic chromatogram of Yunshi Ganmao mixture is with high sensitivity,good reproducibility,simple and convenient operation.By comparing the similarity between the tested samples and the HPLC characteristic chromatogram of Yunshi Ganmao mixture,and combining with stoichiometry,the quality of Yunshi Ganmao mixture can be described and evaluated as a whole to ensure the uniformity of product quality.