HPLC指纹图谱、Q-TOF/MS定性及多成分定量相结合的大黄饮片质量评价研究

Quality evaluation of rhubarb pieces by HPLC fingerprint, Q-TOF/MS qualitative and multi-component quantitative analysis

目的建立更全面的大黄饮片质量评价方法。方法 HPLC法建立35批大黄饮片指纹图谱,并生成对照指纹图谱,标定共有峰,并进行相似度评价;Q-TOF/MS对共有峰进行鉴定,对通过对照品比对确认的13个蒽醌类成分进行定量测定。结果大黄饮片指纹图谱标定共有峰45个,包括蒽醌类成分17个(其中芦荟大黄素-8-O-葡萄糖苷、大黄酸-8-O-葡萄糖苷、大黄素-1-O-葡萄糖苷、大黄酚-1-O-葡萄糖苷、大黄酚-8-O-葡萄糖苷、芦荟大黄素-3-羟甲基-O-葡萄糖苷、大黄素-8-O-葡萄糖苷、大黄素甲醚-8-O-葡萄糖苷8个结合型蒽醌及芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚5个游离型蒽醌通过对照品比对确认)、蒽酮类成分2个(番泻苷B和番泻苷A,均通过对照品比对确认)、鞣质类成分17个(其中没食子酸、儿茶素、表没食子儿茶素没食子酸酯、表儿茶素、表儿茶素没食子酸酯通过对照品比对确认)、二苯乙烯类成分2个[白藜芦醇4′-O-葡萄糖苷和白藜芦醇4′-O-β-D-(6″-O-没食子酰)-葡萄糖苷,均通过对照品比对确认]、苯丁酮类成分4个[其中4′-羟基苯基-2-丁酮-4′-O-β-D-(2″-O-没食子酰-6″-O-对羟基桂皮酰)-葡萄糖苷通过对照品比对确认]、色原酮类成分2个、萘类成分1个;35批大黄饮片中13个蒽醌类成分含量测定结果差异较大。结论建立的方法可用于大黄饮片质量评价。

Objective To establish a more comprehensive quality evaluation method for rhubarb pieces.Methods Fingerprints of 35 batches of rhubarb pieces were determined by HPLC,the control fingerprint was established,the common peaks were calibrated and the similarity was evaluated.Components of common peaks were identified by Q-TOF/MS and the content of 13 anthraquinones confirmed by reference substance was determined.Results There were 45 common peaks in the fingerprint of rhubarb pieces,including 17 anthraquinones(among them,eight bound anthraquinones of aloe-emodin-8-O-glucoside,rhein-8-O-glucoside,emodin-1-O-glucoside,chrysophanol-1-O-glucoside,chrysophanol-8-O-glucoside,aloe-emodin-3-hydroxymethyl-O-glucoside,emodin-8-Oglucoside,physcion-8-O-glucoside and five free anthraquinones of aloe-emodin,rhein,emodin,chrysophanol,physcion were confirmed by reference substance),two anthrones(sennoside B and sennoside A,confirmed by reference substances),17 tannins(among them,gallic acid,catechin,epigallocatechin gallate,epicatechin and epicatechin gallate were confirmed by reference substance),two stilbenes[resveratrol 4′-O-glucoside and resveratrol 4′-O-β-D-(6″-O-galloyl)-glucoside,confirmed by reference substances],four butyrophenone[among them,4-(4′-hydroxyphenyl)-2-butanone 4′-O-β-D-(2″-O-galloyl-6″-O-p-hydroxy-cinnamyl)glucoside,confirmed by reference substances],two chromones and one naphthalenes.The content of 13 anthraquinones in 35 batches of rhubarb pieces was quite different.Conclusion The established method can be used to evaluate the quality of rhubarb pieces.