The Establishment Of UPLC-MS/MS Method For Determination Of Fenbendazole,Oxfendazole,Fenbendazole Sulfone And Ivermectin And Their Pharmacokinetics In Pig Plasma

Fenbendazole belongs to benzimidazole drugs,was mainly used to treat gastrointestinal nematode in clinical veterinary.Oxfendazole and Fenbendazole Sulfone are the main metabolites of Fenbendazole in animal body.Ivermectin is a kind of avermectin drugs,which mainly used in the treatment of digestive tract nematode infection.The combination of Fenbendazole and Ivermectin enlarges the repellent spectrum and improves the one-time repellent effect.Therefore,a method for determination of Fenbendazole,Oxfendazole,Fenbendazole Sulfone and Ivermectin was established using UPLC-MS/MS simultaneously,the concentration of four drugs at different time points in pig plasma has been determined,and Pharmacokinetic parameters and pharmacokinetic time curves of four drugs in three different experimental groups were obtained,providing scientific reference for rational combination of Fenbendazole and Ivermectin in clinic.In the study of establishment of simultaneous determination of Fenbendazole,Oxfendazole,Fenbendazole and Ivermectin in pig plasma using UPLC-MS/MS method,the detection limits of Fenbendazole,Oxfendazole,Fenbendazole and Ivermectin in pig plasma were 0.05 ng/mL,0.05 ng/mL,0.05 ng/mL and 0.01 ng/mL respectively,and the limits of quantification were 0.1 ng/mL,0.1 ng/mL,0.1 ng/mL and 0.2 ng/mL respectively.The recovery rates at three different levels of Fenbendazole,Oxfendazole,Fenbendazole and Ivermectin in pig plasma were 89.42%⁹4.33%,79.05%⁹2.28%,86.19%⁹2.29%and 82.09%⁸8.40%respectively.The linear equation were y=26321.7x+14055.2,y=12348.8x+11594.9,y=13900.8x+7230.8 and y=424.7x-188.7 respectively.The correlation coefficients r were 0.9999,0.9994,0.9995 and 0.9997 respectively.In the study of pharmacokinetics of Fenbendazole,Oxfendazole,Fenbendazole and Ivermectin in pig plasma.30 healthy pigs weighted 20±2 kg,was divided into A,B,C 3 groups,each group of 10,group A compound preparation,group B Fenbendazole and group C Ivermectin were set up.Blood was collected at different time intervals from 0 to 21 d after administration.The concentrations of the 4drugs in the three groups were determined by the UPLC-MS/MS method of the established method Fenbendazole,Oxfendazole,Fenbendazole Sulfone and Ivermectin in pig plasma.After the measured datas was analyzed with WinNonlin 6.1 in non-compartment model model software,the pharmacokinetic parameters of three experimental groups were obtained.The pharmacokinetic parameters of the Fenbendazole Ivermectin group were as follows:The Cmax of FBZ,OFZ,FBZSO2and IVM were like these separately:58.12±20.19μg/L;752.31±129.44μg/L;593.60±138.38μg/L and39.39±16.71μg/L.Tmax 9.30±0.95 h;20.40±5.80 h;24.40±1.26 h and 16.80±6.20 h.AUC710.61±202.31 h·μg/L;15265.36±3568.44 h·μg/L;9751.44±3108.63 h·μg/L and 2409.643±229.08h·μg/L.T_1/2/2 4.96±1.62 h;2.24±0.61 h;7.49±5.67 h and 186.90±71.10 h.The pharmacokinetic parameters of Fenbendazole group were as follows:The Cmax of FBZ,OFZ,FBZSO2 were like these separately:50.23±11.34μg/L;418.80±117.72μg/L;213.03±115.16μg/L.Tmax 7.80±2.10 h;12.90±6.01 h;22.80±3.79 h.AUC 588.07±187.50 h·μg/L;7823.52±3096.71 h·μg/L;3563.48±1693.75h·μg/L.T_1/2/2 3.60±1.42 h;4.50±1.38 h;3.32±3.59 h.The pharmacokinetic parameters of Ivermectin group were as follows:The Cmax of IVM was 32.68±5.41μg/L.Tmax 12.10±4.91 h;AUC3575.27±660.81 h·μg/L.T_1/2/2 114.75±50.30 h.The test results showed that:The Cmax of each drug component in Fenbendazole Ivermectin powder group was larger than that in Fenbendazole group and Ivermectin group.The Tmax and Cmax of each drug component in Fenbendazole Ivermectin powder group was higher than that in Fenbendazole group and Ivermectin group.The AUC of each component in the Fenbendazole Ivermectin powder group was larger than that in the Fenbendazole group.The T_1/2of FBZ,FBZSO2 and IVM in Fenbendazole Ivermectin powder group were higher than those in Fenbendazole and Ivermectin.By comparing the pharmacokinetic parameters,it concluded that the pharmacokinetics of Fenbendazole Ivermectin group was better than that of Fenbendazole group and Ivermectin group,The effect of Fenbendazole Ivermectin powder group was better than that of Fenbendazole powder group and Ivermectin powder group,the combination of the two drugs enhanced the effect of expelling parasite.