文冠果壳苷大鼠体内绝对生物利用度和体外代谢研究

Study on Absolute Bioavailability of Xanthoceraside in Rat and in Vitro Metabolic Behavior

目的 采用LC-MS/MS测定大鼠血浆中文冠果壳苷的浓度,研究其在大鼠体内的口服绝对生物利用度和体外代谢行为。方法 大鼠分别灌胃文冠果壳苷40 mg·kg^(–1)和尾静脉注射文冠果壳苷2 mg·kg^(–1)后,不同时间点采集血样。采用LC-MS/MS分析方法测定大鼠血浆中文冠果壳苷的含量,运用DAS 3.3.2软件计算相关的药动学参数,并通过肠道菌群和肝S9体外孵化试验研究文冠果壳苷的代谢行为。结果 应用于经胃和静脉给药的文冠果壳苷线性浓度范围分别为1.56~100μg·L^(–1)和200~40 000μg·L^(–1),专属性、精密度、准确度、基质效应、提取回收率以及稳定性均符合生物样本分析要求。灌胃给药后大鼠血浆中文冠果壳苷的ρ_(max)、AUC_(0-t)分别为(38.33±33.21)μg·L^(–1)和(100.77±28.85)μg·h·L^(–1);静注给药后大鼠血浆中文冠果壳苷的ρ_(max)、AUC_(0-t)分别为(24 428.01±10 386.18)μg·L^(–1)和(17 355.24±10 498.71)μg·h·L^(–1);文冠果壳苷在大鼠体内的绝对生物利用度为0.028%。与孵化前样品相比,肠道菌群培养48 h和肝S9培养2 h后,文冠果壳苷分别降解为孵化前的90.7%和99.3%。结论 建立的LC-MS/MS分析方法可应用于文冠果壳苷在大鼠体内的药动学研究。文冠果壳苷在大鼠体内的口服绝对生物利用度仅为0.028%,这可能是由于极低的胃肠道吸收引起的。

OBJECTIVE To establish a method to determine the concentration of xanthoceraside(XAN) in rat plasma by LC-MS/MS,and to study its oral absolute bioavailability and metabolic behavior in vitro in rats.METHODS The blood sample was collected at different time points after intragastric administration of XAN at 40 mg·kg^(-1) and intravenous administration of XAN at 2 mg·kg^(-1).XAN in plasma sample was analyzed by the validated LC-MS/MS.DAS 3.3.2 software was used to calculate relevant pharmacokinetic parameters,the metabolic behavior of XAN was studied by intestinal microflora and liver S9 incubation test in vitro.RESULTS The linear concentration ranges of XAN were 1.56-100μg·L^(-1) and 200-40 000μg·L^(-1),which were applied to the determination after intragastric and intravenous administration separately.The specificity,precision,accuracy,matrix effect,extraction recovery and stability of the methods were consistent with the requirements for biological sample analysis.The pharmacokinetic parametersρ_(max) and AUC_(0-t )after oral administration were (38.33±33.21)μg·L^(-1 )and (100.77±28.85)μg·h·L^(-1),respectively;and theρ_(max) and AUC_(0-t) after intravenous injection were (24 428.01±10 386.18)μg·L^(-1) and (17 355.24±10 498.7)μg·h·L^(-1),respectively.The absolute bioavailability of XAN in rat in vitro was 0.028%.In the incubation experiment,XAN remained 90.7%and 99.3%compared with the pre-incubation sample after 48 h of intestinal flora incubation and 2 h of liver S9incubation.CONCLUSION The LC-MS/MS analytical method established is suitable for the pharmacokinetic study of the XAN in rats.The oral absolute bioavailability of XAN is 0.028%,which might be caused by the low absorption in gastrointestinal tract.