小鼠肠道细菌对大承气汤中番泻苷A的高效液相色谱测定及代谢研究

High-performance liquid chromatographic determination and metabolic study of sennoside a in daiokanzoto by mouse intestinal bacteria.

作者信息

Takayama Kento, Matsui Emi, Kobayashi Takeshi, Inoue Hirofumi, Tsuruta Yasuto, Okamura Nobuyuki

机构信息

Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, Japan.

出版信息

Chem Pharm Bull (Tokyo). 2011;59(9):1106-9. doi: 10.1248/cpb.59.1106.

DOI:10.1248/cpb.59.1106

PMID:21881253

Abstract

Daiokanzoto (DKT, combination of rhubarb and glycyrrhiza), a Kampo medicine, is clinically effective for constipation. Sennoside A is well known to induce diarrhea. Sennoside A is a prodrug that is transformed into an active metabolite, rheinanthrone, by intestinal bacteria. In this study, we investigated the effects of glycyrrhiza on the activity of sennoside A metabolism in intestinal bacteria using mouse feces. A high-performance liquid chromatography (HPLC) method for the determination of sennoside A in incubation mixture of DKT with mouse feces was established. The retention time of sennoside A was 9.26±0.02 min with a TSKgel ODS-80TsQA column by linear gradient elution using a mobile phase containing aqueous phosphoric acid and acetonitrile and detection at 265 nm. We found that the activity of sennoside A metabolism in intestinal bacteria was significantly accelerated when glycyrrhiza, liquiritin or liquiritin apioside coexisted with sennoside A, whereas that of glycyrrhizin was not altered. This method is applicable for determination of the activity of sennoside A metabolism by anaerobic incubation of DKT with mouse feces.

摘要

大承气汤（DKT，大黄与甘草的组合），一种汉方药物，临床上对便秘有效。番泻苷A以引起腹泻而闻名。番泻苷A是一种前体药物，通过肠道细菌转化为活性代谢物大黄蒽酮。在本研究中，我们使用小鼠粪便研究了甘草对肠道细菌中番泻苷A代谢活性的影响。建立了一种高效液相色谱（HPLC）方法，用于测定DKT与小鼠粪便孵育混合物中的番泻苷A。使用含磷酸水溶液和乙腈的流动相通过线性梯度洗脱，在265nm处检测，番泻苷A在TSKgel ODS-80TsQA柱上的保留时间为9.26±0.02分钟。我们发现，当甘草、甘草苷或甘草苷芹菜糖苷与番泻苷A共存时，肠道细菌中番泻苷A的代谢活性显著加快，但甘草酸的代谢活性没有改变。该方法适用于通过DKT与小鼠粪便厌氧孵育来测定番泻苷A的代谢活性。