Pharmacokinetics Research Of Rrotopine In Broiler Chickens

Macleaya cordata widely distributed in northwest and southwest of China,Southeast Asia,North America and Europe,which has more than 1,000 years of medication history in China.Macleaya cordata had been developed to many different products,widely used in poultry disease treatment,feed additives,biological insecticide,energy regulation and other fields with its antibacterial,insecticidal,anti-inflammatory,regulating intestinal flora and other pharmacological effects.Macleaya cordata contains aboudant protopine-type alkaloids,mainly protopine?PRO?and allocryptopine?ALL?,and has been developed into an extract products-Bopu San,used in the treatment of laying hens salpingitis.At present now,the laboratory has established the method for the simultaneous determination of PRO,ALL,Sanguinarine?SA?and Chelerythrine?CHE?in rat plasma by UPLC-ESI-MS/MS and explored the pharmacokinetics of total topine alkaloids from Macleaya cordata extracts in the rat plasma.But there was no report about the pharmacokinetic study of the total protopine alkaloids from Macleaya cordata extracts in its target animals.This research was carried out in order to make good use of protopine-type alkaloids,and make a technical preparation for the subsequent evaluation of new drugs safety.The major work of this paper was as follws:1.The UPLC-ESI-MS/MS method for the simultaneous determination of PRO,ALL,SAN and CHE in rat plasma was reviewed and verified by using the yellow broiler chicken plasma.The results of methodological validation showed:the specificity of this method was well.The standard curve of the four alkaloids was well when the linear range was 1.56ng/mL-400 ng/mL?r²?0.997?,and the relative deviation,relative standard deviation,extraction recoveries,matrix effects and stabilities of quality control samples were well-validated.2.The stability of PRO,ALL,SAN and CHE in different pH buffers,artificial gastric juice and artificial intestinal juice were investigated by in vitro experiments.The results showed:The recovery rate of protopine and allocryptopine were ranging from 97.16%to101.89%in the environment of acidic,neutral,alkalinity media,artificial gastric juice and artificial intestinal juice for 8 h.The recovery rate of sanguinarine was48.81%⁶0.53%in the environment of pH=6.8 phosphate buffer solution?PBS?,water,pH=7.8-8.0 PBS and artificial intestinal juice for 8 h.The recovery rate of chelerythrine was81.21%⁹3.48%in the environment of pH=5.8 PBS,pH=6.8 PBS,water and artificial intestinal juice for 8 h;while in the environment of pH=7.8-8.0 PBS for 8 h,the recovery rate of chelerythrine was 57.43%.And we concluded that the stability of protopine and allocryptopine was not affected by pH.While along with the increase of pH,the stability of sanguinarine and chelerythrine was reduced.The stability of four alkaloids in Macleaya cordata under the simulated gastrointestinal environment accorded with the environment of different pH media.3.The plasma protein binding rate of PRO and ALL was investigated by balanced dialysis.The method was applied the blank plasma in a dialysis bag to the dialysate solution containing a concentration of alkaloid.And the plasma was equilibrated at 37?and 120r/min for 24 h.At the same time,we compared the plasma protein binding rate of PRO single standard,ALL single standard and PRO,ALL mixed standard.The results showed:The plasma protein binding rate of PRO was 39.72±3.99%,which was a moderate degree of combination.And the plasma protein binding rate of ALL was 77.12±2.87%,which was a high degree of combination.PRO and ALL plasma protein binding rates were not concentration-dependent,and there was no competition between PRO and ALL.4.The contents of alkaloids in Bopu San were measured by using High performance liquid chromatography.PRO 1.10%,ALL 0.40%,SAN 0.05%,CHE 0.02%.5.The animals were orally administered Bopu San at a dose of 240 mg/feathers?PRO2.64 mg/feather,ALL 0.96 mg/feather?and intravenous administered protopine alkaloids?PRO 1.2?g/feather,ALL 0.6?g/feather?,the blood was collected from the wing vein into heparinized centrifuge tubes at different time.Pharmacokinetic studies have shown that the Cmax of PRO and ALL were 22.09 and 28.33?g/L;Tmax was 0.67 and 0.675 h;MRT_?0-??was 14.96 and 2.91 h;the elimination half-life t_1/2 was 17.68 and 2.09 h,respectively.Moreover,the concentration of PRO and ALL in the plasma was 3.1 and 6.513?g/L after 5min of intravenous injection of mixed solution including PRO and ALL,respectively.The plasma PRO and ALL were almost undetected after intravenous injection for 1 h and 1.5 h,respectively.Therefore,these results suggested that the absorption and elimination of PRO and ALL in broilers were fast,and the absorption rate was low.The time of PRO in vivo was longer,and the residence time of ALL was shorter.The absolute bioavailability of PRO and ALL were 2.07%and 1.19%respectively,which indicated that the bioavailability of alkaloids in broilers was low.The retention time of PRO in broilers was longer,but the retention time of PRO in broilers was shorter.The absolute bioavailability of PRO and ALL were 2.07%and 1.19%respectively,which indicated that the bioavailability of alkaloids in broilers was low.