DU Huan, Xu Xin-Mei, Xu Tong, Li Qi, Zhao Cheng-Cheng, Yi Huan, Fan Gang
College of Pharmacy, Chengdu University of Traditional Chinese Medicine Chengdu 611137, China.
College of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine Chengdu 611137, China.
Zhongguo Zhong Yao Za Zhi. 2020 Jan;45(2):418-424. doi: 10.19540/j.cnki.cjcmm.20190830.203.
To elucidate the absorption and metabolism of alkaloids in Berberis kansuensis in vivo, a high performance liquid chromatography-triple quadrupole mass spectrometry(HPLC-QqQ-MS) method was developed to qualitatively and quantitatively analyze the absorption components in rat serum in multiple-reaction monitoring mode. The mobile phase consisted of 0.1% formic acid and acetonitrile with a gradient elution mode. In addition, to investigate the effects of gut microbiota on five absorbed components of B. kansuensis in rat serum, diabetic rat and pseudo germ-free diabetic rat models were established, and partial least squares discriminant analysis and One-way ANOVA were used to study the content differences of five components among different groups. In this study, a HPLC-QqQ-MS method for quantitative analysis of five components in rat serum after oral administration of B. kansuensis was established for the first time. It was found that there were differences in the five constituents in rat serum between different groups. By comparing the normal group with the diabetic model group, we found that the absorption and metabolism capacities of berberine and magnoflorine were different under the health and pathological conditions. It was also found that the serum levels of berberine, magnoflorine and jatrorrhizine in pseudo germ-free diabetic rats were significantly lower than those in diabetic rats, indicating that gut microbiota plays an important role in the metabolism of alkaloids of B. kansuensis in vivo. These results provide a good reference for clarifying the active ingredients of B. kansuensis in the treatment of diabetes.
为阐明甘肃小檗中生物碱在体内的吸收和代谢情况，建立了一种高效液相色谱 - 三重四极杆质谱联用（HPLC - QqQ - MS）方法，以多反应监测模式对大鼠血清中的吸收成分进行定性和定量分析。流动相由0.1%甲酸和乙腈组成，采用梯度洗脱模式。此外，为研究肠道微生物群对大鼠血清中甘肃小檗五种吸收成分的影响，建立了糖尿病大鼠和伪无菌糖尿病大鼠模型，并采用偏最小二乘判别分析和单因素方差分析研究不同组间五种成分的含量差异。本研究首次建立了一种HPLC - QqQ - MS方法，用于定量分析甘肃小檗口服给药后大鼠血清中的五种成分。发现不同组大鼠血清中的五种成分存在差异。通过比较正常组和糖尿病模型组，发现健康和病理条件下小檗碱和木兰碱的吸收和代谢能力不同。还发现伪无菌糖尿病大鼠血清中小檗碱、木兰碱和药根碱的水平显著低于糖尿病大鼠，表明肠道微生物群在甘肃小檗生物碱的体内代谢中起重要作用。这些结果为阐明甘肃小檗治疗糖尿病的活性成分提供了良好的参考。
