蜗牛酶改善芍药苷在比格犬体内肠道水解和生物利用度研究

The study of snailse in improving hydrolysis and bioavailability of paeoniflorin in intestinal of beagle dogs

目的分析蜗牛酶改善芍药苷在比格犬体内肠道水解和生物利用度的情况。方法取12只雄性比格犬,分成A和B两组,每组6只,按20 mg/kg的剂量用芍药苷给比格犬灌胃。A组直接芍药苷灌胃,B组采用含蜗牛酶和芍药苷的复合物肠溶胶囊,采用液相色谱-串联质谱法监测并计算给药后0.25 h、0.5 h、1 h、1.5 h、2 h、2.5 h、3 h、4 h、6 h、8 h、12 h及24 h各时间点的血药浓度、药动学参数、达峰浓度(Cmax)和达峰时间(Tmax)、浓度-时间曲线下面积(area under concentration time curve,AUC0-t),对芍药苷在比格犬体内的药物动力学进行评价。结果芍药苷的体内浓度为0.19~970.00 ng/ml时,线性关系良好。定量限为0.19 ng/ml,低、中及高水平的芍药苷血浆样品回收率为83.45%~92.34%,日间精密度及日内精密度均小于9.34%。比格犬口服芍药苷Cmax=(717.03±76.44)ng/ml,Tmax=(1.00±0.29)h,AUC0-t=(1621.11±98.99)ng·h/ml;口服肠溶胶囊Cmax=(584.93±38.85)ng/ml,Tmax=(2.58±0.20)h,AUC0-t=(2656.87±165.34)ng·h/ml,与单用芍药苷相比,口服肠溶胶囊时AUC增大,Tmax延长,平均滞留时间延迟,差异有显著统计学意义(P<0.01)。结论液相质谱联用方法快速、准确,且重复性好,适用于比格犬中芍药苷的药物动力学研究。含蜗牛酶和芍药苷的肠溶胶囊较单纯芍药苷在一定程度上可提高芍药苷的相对生物利用度。

Objective To investigate whether snailase enzyme can improve intestinal hydrolysis and bioavailability of paeoniflorin in beagle dogs.Methods A total of 12 male beagles were divided into group A and group B,and each group consisted of 6 beagles.Paeoniflorin was given to beagle dogs by gavage at a dose of 20 mg/kg.In group A,paeoniflorin was infused directly into the stomach.In group B,snail enzyme and paeoniflorin compound enteric coated capsules were used.The pharmacokinetics of paeoniflorin in beagle dogs was evaluated.The plasma concentration,pharmacokinetic parameters,peak concentrationin(Cmax),peak time(Tmax)and area under concentration time curve(AUC0-t)were measured and calculated at 0.25 h,0.5 h,1 h,1.5 h,2 h,2.5 h,3 h,4 h,6 h,8 h,12 h and 24 h after administration by liquid chromatography tandem mass spectrometry(LC-MS/MS).Results Within the area 0.19~970 ng/ml,paeoniflorin presented with good linear relationship.The limit of quantification was 0.19 ng/ml.The recovery of paeoniflorin plasma samples at low,medium and high levels was 83.45%~92.34%,and the intra-and inter-day precisions were less than 9.34%.The Cmax of group A was(717.03±76.44)ng/ml,Tmax was(1.00±0.29)h,AUC0-t was(1621.11±98.99)ng·h/ml.The Cmax of group B was(584.93±38.85)ng/ml,Tmax was(2.58±0.20)h,AUC0-t was(2656.87±165.34)ng·h/ml.Compared with paeoniflorin alone,the AUC0-t increased,Tmax and verage retention time of enteric coated capsules increased significantly.The difference was statistically significant(P<0.01).Conclusion LC-MS/MS is a rapid,accurate and reproducible method,which is suitable for the pharmacokinetic study of paeoniflorin in beagle dogs.Compared with paeoniflorin alone,enteric coated capsules containing snail enzyme and paeoniflorin can improve the relative bioavailability of paeoniflorin to some extent.