UPLC法测定熟大黄鞣质和蒽醌类成分在不同介质中的溶出差异  

Determination on tannins and anthraquinones of Prepared Rhubarb in different dissolution mediums by Ultra Performance Liquid Chromatography

作　　者：周灿平[1,2] 王伽伯[1] 肖小河[1] 夏新华[2] 张学儒[1,2] 赵海平[1] 

机构地区：[1]解放军中药研究所解放军第三0二医院,北京100039 [2]湖南中医药大学药学院,长沙410208

出　　处：《中华中医药杂志》2010年第11期1788-1792,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家科技部“重大新药创制”科技重大专项(No.2009ZX09502-022);国家科技部中医药行业科研专项(No.200807020)~~

摘　　要：目的:通过模拟人体胃肠环境,比较不同溶出介质中熟大黄鞣质及蒽醌类成分的溶出量差异,从体外溶出的角度探讨影响中药量-效关系的可能因素。方法:取同一批生大黄炮制成熟大黄,选取4种不同溶出介质,采用浆法进行熟大黄体外溶出试验,以UPLC法测定大黄直接溶出和水解后鞣质及蒽醌类成分的含量,色谱柱为Waters Acquity BEHC18柱(50mm×2.1mm,1.7μm),流动相为0.1%磷酸-水(A)-甲醇(B)线性梯度洗脱:0-1.5min,100%A;1.5-2.0min,100%-95%A;2.0-3.5min,95%-86%A;3.5-5.0min,86%-72%A;5.0-8.0min,72%-64%A;8.0-10.0min,64%-52%A;10.0-13.0min,52%-30%A;13-16.0min,30%-16%A。检测波长254nm,流速0.75mL/min,柱温35℃。结果:①生大黄炮制成熟大黄后,已检测不到儿茶素和表儿茶素。②鞣质和蒽醌类成分分别在30、60min后基本达到最大溶出。③水解前后均能检测到没食子酸,且水解后测得含量增加。④水解前仅大黄酸在人工肠液和磷酸盐缓冲液中能检测到,且其在磷酸盐缓冲液中的溶出量大于在人工肠液中的溶出量。⑤同一蒽醌衍生物在人工胃液中含量变化较慢,在水、人工肠液和磷酸盐缓冲液中含量变化相对较快,同时,比较1h内的5种蒽醌含量变化可知,大黄素甲醚在各介质中变化最慢,含量最低,其次是大黄素。结论:不同溶出介质对鞣质和蒽醌类成分的溶出量有重要影响,没食子酸的溶出量大于蒽醌类成分(结合蒽醌),蒽醌类成分在人工肠液和磷酸盐缓冲液中溶出较快,说明小肠和大肠可能是蒽醌溶出的主要部位。Objective： The diversity of dissolution on tannins and anthraquinones of rhubarb in different mediums was compared via simulating human gastrointestinal environment, in order to investigate the potential influential factors of the doseeffect relationship of Chinese Materia. Methods： Prepared rhubarb was processed from the same batch, then the dissolution tests were carried out using sirup method in four dissolution mediums, ultra performance liquid chromatography （UPLC） was chosed to determine the content of tannins and anthraquinones before and after hydrolyses. The compounds were separated on a reversed phase column （Acquity BEH UPLC C 18 100mm×2.1mm, 1.7μm） with a linear gradient elution of 0.1% formic acid in water （A） and methanol （v/v, B）. The gradient elution procedure of the UPLC method was： 0-1.5min, 100%A; 1.5-2.0min, 100-95%A; 2.0-3.5min, 95-86%A; 3.5-5.0min, 86-72%A; 5.0-8.0min, 72-64%A; 8.0-10.0min, 64-52%A; 10.0-13.0min, 52-30%A; 13-16.0min, 30-16%A. The mobile phase flow rate was 0.75 ml/min and monitored at 254nm with the column temperature at 35℃. Results： ①Compared with the crude rhubarb, catechin and epicatechin couldn＇t be determined in prepared rhubarb. ② Tannins and anthraquinones reached their max dissolution at 30 minutes and 60 minutes respectively. ③Gallic acid could be determined before and after hydrolyses, more over, the content of gallic acid was increased after hydrolyses. ④Only rhein could be determined in both artificial intestinal juice and phosphate buffer, and the content in phosphate buffer was higher than in artificial intestinal juice at the same time. ⑤The changes of content of the same anthraquinones was relatively faster in other three mediums than in artificial gastric juice, meanwhile, physcin showed the lowest changing speed within one hour in each mediums followed by emodin. Conclusions： The dissolution of tannins and anthraquinones of rhubarb were greatly affected by the dissolution mediums. It was showed that the dissolution

关 键 词：熟大黄 体外溶出 蒽醌 鞣质 UPLC 

分 类 号：R284.1[医药卫生—中药学]