目的: 采用液相色谱-三重四极杆质谱（LC-MS/MS）技术对三叶苷在Sprague-Dawley（SD）大鼠体内药动学参数、口服生物利用度以及体内代谢物进行系统分析。方法: 采用ACQUITY UPLC BEH C18色谱柱（50 mm×2.1 mm,1.7 µm）,以0.1%甲酸（A）-乙腈（B）为流动相,梯度洗脱,流速0.3 mL · min-1,柱温40 ℃,进样量2 μL。采用电喷雾离子源,负离子化方式,离子源温度150 ℃,毛细管电压-3.0 kV,去溶剂气温度500 ℃,去溶剂气流量750 L · h-1, 锥孔气体积流量150 L · h-1, 多反应监测模式（MRM）检测。三叶苷分别灌胃和静脉注射给予大鼠,收集血浆、尿液、粪便样品,甲醇沉淀蛋白后测定药物浓度,应用药动学软件和代谢物分析鉴定软件对药动学参数、代谢物进行系统分析。结果: 分别灌胃和静脉注射给予SD大鼠三叶苷(100 mg · kg-1)后,测得AUC0-t分别为（423.98±295.42）ng · h · mL-1和（90 894.75±25 472.44）ng · h · mL-1;口服生物利用度为0.46%;Cmax分别为（203.83 ± 25.88）ng · mL-1和（181 814.90±113 461.60）ng · mL-1;口服半衰期1.65 h,静脉注射半衰期3.82 h;三叶苷在肠道代谢为根皮素,并在体内发生脱糖苷化、甲基化、脱氧、水解等生物转化。结论: 对三叶苷大鼠体内药动学和体内代谢进行了初步研究,可为药效研究及后续制剂研发提供参考。
Objective: To systematically analyze the pharmacokinetic parameters, oral bioavailability and in vivo metabolites of trilobatin in Sprague-Dawley (SD) rats using liquid chromatography - triple quadrupole mass spectrometry (LC-MS/MS). Methods: The chromatographic conditions were performed on an ACQUITY UPLC BEH C18 column (50 mm×2.1 mm, 1.7 µm) with 0.1% formic acid (mobile phase A) and acetonitrile (mobile phase B) as the mobile phase. A gradient elution program was carried out with an accompanying flow rate of 0.3 mL · min-1,a column temperature of 40 ℃, and an injection volume of 2 μL. The mass spectrometry conditions comprised an electrospray ion source in conjunction with negative ionization mode, with an ionogenic temperature of 150 ℃, acapillary voltage of -3.0 kV, and a desolvation-gas flow temperature of 500 ℃. The desolvation-gas flow rate was set at 750 L · h-1, and the conical pore gas volumetric flow rate was fixed at 150 L · h-1. The analysis was conducted in multiple reaction monitoring mode. Trilobatin was given to rats via gavage and intravenous injection, respectively. Plasma, urine and fecal samples were collected, and the drug concentration was determined after methanol precipitation of proteins. Pharmacokinetic parameters and metabolites were analyzed by pharmacokinetic software and metabolite analysis and identification software. Results: Following the administration of trilobatin to SD rats at a dose of 100 mg · kg-1 via gavage and intravenous injection, respectively. The area under the curve (AUC0-t) was found to be (423.98 ± 295.42) ng · h · mL-1 and (90 894.75 ± 25 472.44) ng · h · mL-1, respectively. The oral bioavailability was determined to be 0.46%; Cmax was (203.83±25.88) ng · mL-1 and (181 814.90±113 461.60) ng · mL-1, respectively. The oral half-life was 1.65 h, while the intravenous half-life was 3.82 h. Trilobatin was metabolized to phloretin in the intestine and underwent further biotransformation in vivo through deglycosylation, methylation, deoxygenation and hydrolysis. Conclusion: The pilot study represents a preliminary investigation into the in vivo pharmacokinetics and metabolism of trilobatin in rats, providing a foundation for further pharmacodynamics research and subsequent formulation development.
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