Study On The Residues Of Total Alkaloids Of B. Rapa In Broiler Chicken Tissues And Organ

Plume Poppy Total Alkaloid is a Chinese veterinary drug product extracted from[Macleaya cordata（Willd）R.Br.],and its main medicinal ingredient is protopine（PRO）and allocryptopine（ALL）,in addition to a small amount of Sanguinarine（SA）and Chelerythrine（CHE）.The product has good effects on bactericidal,anti-inflammatory,growth-promoting and intestinal flora adjustment for animals such as pigs and chickens.So far,there have been no reports on the residue detection methods and related residue studies of Bop total alkali premix in chicken tissues.Therefore,in order to better guide the application of Plume Poppy Total Alkaloid in clinical veterinary.To make a forward-looking safety evaluation of this product,this paper developed the detection method of PRO,ALL,SA and CHE in broiler tissues,and the pharmacokinetics and residual elimination of the total alkaloids in the plasma of white feather broilers were studied..In this experiment,tetrahydropalmatine（TET）was used as an internal standard to establish a high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS）method for the determination of PRO,ALL,SA and CHE in broiler tissues,and the corresponding method validation was completed.The results of the methodological study showed that PRO,ALL and CHE were linear in the concentration range of 0.5-200μg/kg（r2≥0.999）,and SA was stable in the concentration range of 2²00 μg/kg.The lower limit of quantitation（LLOQ）of PRO,ALL and CHE is 0.5 μg/kg,the lower limit of quantitation of SA is 2 μg/kg,the recovery of all target analytes is 86.61%¹11.17%,and the precision and accuracy are ≤15 %.The method is simple in operation,economical and time-saving,and meets the requirements of methodological investigation in terms of specificity,accuracy and precision,method recovery rate,matrix effect and stability.The detection methods of PRO,ALL,SA and ALL established in our laboratory in chicken plasma and the method of detection and verification of chicken tissue mass spectrometry established in this paper were successfully applied to the pharmacokinetics of PRO,ALL,SA and CHE in broilers and the elimination rules tissues.Mix orally in the feed with a low dose(50 mg·kg^-1),medium(200 mg·kg^-1),and high(800 mg·kg^-1)of Plume Poppy Total Alkaloid.With standardized feeding from the age of one day to 36 days of age,and 12 hours after the fasting,the same amount of feed is given at the same time,and the blood is collected in the pterygopalatine sinus at different time points.m L.The pharmacokinetic parameters indicated that PRO and ALL absorbed quickly and eliminated quickly in chickens,and the blood concentration was below the limit of quantification（0.78ng/ml）after 10 hours of administration.After the blood collection is completed,the broiler enters a one-week washout period,during which the feed of the corresponding dose group is continued.After the blood collection is completed,the broiler enters a one-week washout period.After feeding for 43 days,samples of blood,heart,liver,kidney,chest muscle,leg muscles,and cecal contents were collected on fasting for 2 hours,on the first day,the third day,the fifth day,and the seventh day of the drug administration.The results showed that the low and medium dose groups of PRO and ALL had lower concentrations in different tissues,but they were below the limit of quantitation after 7 days of drug withdrawal.The residual concentration of liver and kidney tissues was the highest,indicating liver and kidney.It is the target tissue of PRO and ALL.The high-dose group still had some residual in the heart,kidney and chest muscle tissue after 7 days of drug taking,and the concentration was0.6¹.16 μg/kg.The residual results demonstrate that long-term feeding of broilers with Plume Poppy Total Alkaloid is safe,indicating that the drug is highly safe.This experiment established a quantitative method for the determination of PRO,ALL,SA and CHE in broiler tissues,and studied the pharmacokinetics of PRO and ALL in broilers and the total alkaloid premix in broiler tissues and organs.The rule of residue elimination provides theoretical guidance and technical support for the clinical use of Plume Poppy Total Alkaloid,and provides a prospective study for the safety evaluation of Plume Poppy Total Alkaloid.