白芍中芍药苷和芍药内酯苷在大鼠血浆中含量测定及药动学研究

摘要

Objective To establish a method for the content determination of paeoniflorin and paeonolide in rat plasma by HPLC-MS/MS; To study its characteristics of pharmacokinetics.Methods HPLC-MS/MS method was adopted. The column was Ultimate RXB-C18 (2.1 mm×100 mm, 3 μm) with mobile phase consisted of acetonitrile- 0.1％ formic acid water solution. Flow rate was 0.3 mL/min; column temperature was 30℃; the volume was 5 μL; detection wavelength was 230 nm. Quantitative ions were m/z 525→120.9 (paeoniflorin and paeonolide), m/z 433→224.8 (geniposide). 8 SD male rats were collected blood 0.5 mL from orbital vein 0, 5, 10, 20, 30, 45, 60, 90, 120, 180, 360, 540, 720 min after intragastric administrationBaishaoDecoction 6 g (medicinal materials)/kg to determine the blood concentration of medicine. DAS V2.1.1 software was employed to calculate pharmacokinetic parameters. Results Good linearity was found in the range of 40–8000 ng/mL and 40–10000 ng/mL for paeoniflorin and paeonolide (r=0.9996,r=0.9995). The intra-day and inter-day RSD were less than 6％. The extraction recoveries were 92.33％–95.43％ and 88.65％–95.62％. The matrix effect values were 99.29％–107.86％ and 93.79％–104.05％.All items of method validation were in line with the requirements. The main pharmacokinetic parameters of paeoniflorin and paeonolide were showed as follows: Cmax of (8.50±2.01) and (4.13±0.72) μg/mL; tmax of (10.00± 1.73) and (20.00±2.11) min; t1/2 of (142.98±30.11) and (127.68±35.74) min.Conclusion The HPLC-MS/MS method established in this study is simple, feasible, accurate, reliable, highly sensitive, and specific, which can be applied to study the pharmacokinetic characteristics of paeoniflorin and paeonolide in rats in vivo.%目的 建立测定大鼠血浆中芍药苷和芍药内酯苷含量的高效液相色谱-质谱联用(HPLC-MS/MS)方法,并用于研究其在大鼠体内的药动学特征.方法 采用HPLC-MS/MS方法,色谱柱为Ultimate RXB-C18(2.1 mm×100 mm,3μm),流动相为乙腈-0.1％甲酸水溶液(30:70,V/V),流速0.3 mL/min,柱温30℃,进样量5μL,检测波长230 nm;定量离子对为:m/z 525→120.9(芍药苷和芍药内酯苷)、m/z 433→224.8(内标物栀子苷).8只雄性SD大鼠,给予6 g原药材/kg白芍汤灌胃,分别于0、5、10、20、30、45、60、90、120、180、360、540、720 min眼眶静脉取血0.5 mL,测定血药浓度,采用DAS V2.1.1药动学软件计算其主要药动学参数.结果 芍药苷和芍药内酯苷分别在40～8000 ng/mL和40～10000 ng/mL范围内线性关系良好(r值分别为0.9996和0.9995),日内和日间精密度RSD均小于6％,提取回收率分别为92.33％～95.43％和88.65％～95.62％,基质效应分别为99.29％～107.86％和93.79％～104.05％,方法学考察均符合要求.主要药动学参数:芍药苷、芍药内酯苷Cmax分别为(8.50±2.01)、(4.13±0.72)μg/mL,tmax分别为(10.00±1.73)、(20.00±2.11)min,t1/2分别为(142.98±30.11)、(127.68±35.74)min.结论 本研究所建立的HPLC-MS/MS方法简单可行、准确可靠、灵敏度高、专属性强,可应用于研究与分析芍药内酯苷和芍药苷在大鼠体内的药动学特征.