玳玳果黄酮降脂提取物体内组织分布规律及特征研究

目的:建立UPLC-MS/MS分析方法同时测定玳玳果黄酮降脂提取物效应组分新橙皮苷和柚皮苷在大鼠10种脏器组织中含量,分布规律及特征。方法:采用UPLC-MS/MS技术建立提取物效应组分新橙皮苷及柚皮苷在大鼠心、肝、脾、肺、肾、脑、胃、小肠、脂质、肌肉组织中的定量分析方法;大鼠给药后分别于0.33,0.67,1,4,8 h的5个时间点,分别摘取以上10种脏器组织,测定脏器组织及血液中效应组分的质量浓度,采用DAS(V 2.0)药动学软件对各样本的药物浓度-时间数据进行房室拟合,并计算不同组织效应组分的药-时曲线下面积(AUC)及平均滞留时间(MRT)。结果:所建立的UPLC-MS/MS定量分析方法具备良好的专属性、标准曲线及线性范围良好、方法准确度与精密度、定量下限均符合有关规定;玳玳果黄酮降脂提取物效应组分在血液中的分布符合一室模型,除肾脏及脑组织外,其余脏器中提取物效应组分的房室特征多为静脉注射的二室模型,柚皮苷在肾脏中的拟合结果为非静脉注射的二室模型,新橙皮苷在脑组织拟合结果为静脉注射的三室模型,给药后8 h各组织中效应组分新橙皮苷及柚皮苷AUC值大小顺序均为小肠>胃>肾>脂质≈脾脏>肺>肌肉>肝>心>脑,效应组分在各脏器中均无明显蓄积;效应组分在血液、肾脏、肝脏中的滞留时间较长,MRT均大于2 h,脂质最短,MRT不足1 h;各脏器中新橙皮苷的药-时曲线下面积约是柚皮苷的3倍,而心、肝、肾中则是3.5,2.1和3.4倍。结论:玳玳果黄酮降脂提取物效应组分在大鼠组织中分布迅速,达峰时间早于血液;效应组分在肠道内消除缓慢,给药8 h后在各脏器中的含量均显著下降且无特异的蓄积部位。研究结果揭示玳玳果黄酮降脂提取物效应组分在大鼠体内的分布特征及规律,为进一步理解玳玳果黄酮降脂提取物在体内的作用靶点及机制奠定了基础。

Study on the patterns and characteristics of the in vivo tissue distribution of Daidai lipid-lowering flavonoid extract

OBJECTIVE To establish a UPLC-MS/MS method for the simultaneous determination of the contents of the active ingredients (neohesperidin and narngin) in 10 kinds of rat organs and tissues and to study the characteristics and patterns of the in vivo tissue distribution of active ingredients. METHODS UPLC-MS/MS was used to establish a quantitative analysis method for the active ingredients (neohesperidin and narngin) in rat heart, liver, spleen, lung, kidney, brain, stomach, small intestine, lipid, and muscle tissue. At 0.33, 0.67, 1, 4, and 8 hours after administration, the 10 organs and tissues were respectively extracted to determine the mass concentration of active components in organ tissues and blood. DAS (V 2.0) pharmacokinetic software was used to perform compartment fitting for drug concentration-time data of each sample, and the area under the concentration-time curve (AUC) and mean retention time (MRT) of different tissue effect components were calculated. RESULTS The established UPLC-MS/MS quantitative analysis method had good specificity, standard curve and linear range. Accuracy, precision and lower limit of quantification were consistent with appropriate standards;. The distribution of the active ingredients of Daidai lipid-lowering flavonoid extract in blood was fitted to a one-compartment model. Apart from kidney and brain tissue, the atrioventricular characteristics of the active ingredients in other organs and tissues could be described as a intravenous two-compartment model. The fitting result of narngin in kidney was a non-intravenous two-compartment model, and the fitting result of neohesperidin in brain tissue was a intravenous three-compartment model. Eight hours after the administration, the order of the AUC values of neohesperidin and narngin in organs and tissues were small intestine > stomach > kidney > lipid ≈ spleen > lung > muscle > liver > heart > brain. No significant accumulation of the active ingredients was observed in all organs and tissues. The retention time of the active ingredients was relatively longer in blood, kidney, and liver with MRT longer than 2 hours. Lipid had the smallest MRT, which was shorter than 1 hour. The area under concentration-time curve values of neohesperidin in organs and tissues were about 3 times of those of narngin. The multiples in heart, liver, and kidney were 3.5, 2.1, and 3.4 respectively. CONCLUSION The whole fate of Daidai lipid-lowering extract of hawthorn fruit is characterized by the rapid distribution of active components in tissues and the earlier peak time than that in blood; the effect components are slowly eliminated in the intestinal tract, and the contents in all organs are significantly decreased after 8 h of administration and there is no specific accumulation site. The results revealed the distribution characteristics and regularity of the effective components of the Daidai lipid-lowering extract in rats, which laid the foundation for further understanding of the in vivo target and mechanism of the Daidai lipid-lowering extract.