LC-MS/MS法测定Beagle犬血浆中曲札茋苷及其药动学研究

Determination of piceatannol 3’-O-glucoside in the plasma of Beagle dogs using LC-MS/MS and its pharmacokinetic research

目的:建立LC-MS/MS法测定Beagle犬血浆中曲札茋苷的浓度,并用该方法研究其在Beagle犬体内的药动学特征。方法:以虎杖苷为内标,血浆样品采用甲醇沉淀的方式分离蛋白并离心取上清进样,色谱柱为Kromasil^(■)C_(18)柱(150 mm×2.1 mm)。以10 mmol·L^(-1)甲酸铵水溶液和乙腈为流动相,梯度洗脱,流速为0.3 mL·min^(-1)。监测的离子对分别为m/z 405.1~243.0(曲札茋苷)和m/z 389.1~226.9(虎杖苷)。Beagle犬采用静脉输注(iv gtt)的方式给予不同剂量的药物,不同时间点取样,测定其血浆中曲札茋苷的浓度。由DAS 2.0计算药动学参数。结果:血浆中曲札茋苷在1.0~200.0 ng·mL^(-1)范围内线性关系良好(r=0.9998),批内和批间精密度(RSD)均<15%。Beagle犬iv gtt给予不同剂量的曲札茋苷后,曲札茋苷的t_(1/2)为(53.3±15.5),(52.1±14.4)和(55.1±16.1)min,AUC_(0→t)为(15358±1815),(52401±3315)和(203419±43778)μg·min·L^(-1)。结论:本方法快速简便、灵敏度高、准确性好,适用于曲札茋苷的血药浓度测定和药动学研究。

Objective: An LC-MS/MS method was established to determine the concentration of piceatannol 3’-O-glucoside in the plasma of Beagle dogs, which was further used for the pharmacokinetic characteristics. Methods: Polydatin was used as the internal standard. The plasma samples were precipitated by methanol. After centrifugation, the supernatant was injected for analysis. A Kromasil^(■) C_(18) column(150 mm×2.1 mm) was used with 10 mmol·L^(-1) ammonium formate aqueous solution and acetonitrile as mobile phases following a programmed gradient elution.The flow rate was set at 0.3 mL·min^(-1). The monitored ion pairs were m/z 405.1~243.0(piceatannol 3’-O-glucoside) and m/z 389.1~226.9(polydatin). Beagle dogs were administered different doses of drugs by intravenous infusion, and the plasma samples were taken at different time points to determine the concentration of piceatannol 3’-O-glucoside. The pharmacokinetic parameters were calculated by DAS 2.0. Results: Piceatannol 3’-O-glucoside in plasma had a good linear relationship in the range of 1.0~200.0 ng·mL^(-1)(r=0.999 8), and the intra-day and inter-day precision(RSD) were both within ±15%. After the Beagle dogs were given intravenous infusion of piceatannol 3’-O-glucoside at different doses, the t_(1/2) values of piceatannol 3’-O-glucoside were(53.3±15.5),(52.1±14.4) and(55.1±16.1) min, and AUC_(0→t) values were(15 358±1 815),(52 401±3 315) and(203 419±43 778) μg·min·L^(-1),respectively. Conclusion: This method is fast, simple, high in lucidity, and good in accuracy. It is suitable for the blood concentration determination and pharmacokinetic study of piceatannol 3’-O-glucoside.