Pharmacokinetic Study Of Eugenol In Carp

Carps cause injury in the areas of long distance transport,breeding,cultivation,surgery,fishing and marketing due to hypoxia and stress.Anesthetics can reduce the casualty rate of fish effectively,so as to improve the value of aquatic products.Therefore,to explore the fish anesthetic which is efficient,economy,low toxicity and no residual harm has become a hot research topic.Eugenol cloves is derived from plants.It is often used as food additives and flavouring agent.It is low prices and high safety.But the detection method of eugenol in fish is not perfect in the current.It still is unable to evaluate the safety through using as a carp narcotic.This hinders its application.To explore the safety of eugenol after anesthesia which use eugenol and the relationship of ugenol anesthesia effect with the concentration of central nervous system.This study carried out establishing HPLC(High Performance Liquid)method determination of eugenol in carp blood and tissue content,the optimal concentration of eugenol anesthesia carp and treatment index in the research,the investigation of the rule of eugenol in carp blood eliminate,eugenol in the research on the laws of the carp in the organization to eliminate and eugenol anesthesia carp PK-PD model;in order to determine whether the eugenol anesthesia carp fish edible safety and to provide support of new drug research and development.Results1.High performance liquid chromatography(HPLC)method was established to detect the contents of eugenol in carp plasma and tissues.The method pretreatment uses methanol,acetonitrile as the extractant and removes the protein,the high speed centrifugation and the filtration form removes the solid particles,and n-hexane is degreased.Proportion of mobile phase was methanol-acetonitrile-water(31:31:38),flow rate was 1 mL/min,ultraviolet absorption was at 280 nm,the temperature of column was35℃and sample amount was 20μL.The results showed that eugenol concentration waswithin 0.041-21.2μg/mL,have a good linear relationship with the peak area ratio,its R2 value was greater than 0.999 and standard curves of tissues were good linear relationship.The average recovery of eugenol in carp tissues was 86.7%-98.1%.The results of precision,repeatability and stability in each tissue experiments showed that RSD was0.005%-2.768%,difference was small.It showed that this method was sensitive,precision,reproducibility,stability and added high recovery rate.It was fully applicable to the carp on the determination of eugenol content in blood and tissues.2.With the condition of water temperature was 24℃and carps which were0.75±0.1kg,eugenol emulsion to dipping bath carps.The optimal anesthetic concentration was 35 mg/l.Median effective dose was 26.374 mg/l.Median lethal dose was 424.384mg/l.The rapeutic index was 16.091 mg/l.Therapeutic index was relatively high.And it showed that the eugenol emulsion was used for anesthesia carps was safty.3.The temperature was 24℃,The period of 0.25 h to 0.5 h,drug concentration in plasma made the biggest change.With the increase of medicated bath concentration,the eliminate of eugenol in carps blood prolonged.When concentration of eugenol emulsion was 20 mg/l,the plasma of eugenol was undetectable at 24 h.When concentration of eugenol emulsion was 35 mg/l,the plasma of eugenol was undetectable at 48 h.When concentration of eugenol emulsion was 75 mg/l,the plasma of eugenol was still detectable at 48 h(The content of eugenol in plasma was 0.060μg/ml).4.The change of eugenol content in plasma:The higher medicated bath concentration was,the higher the eugenol content of the prime was.The temperature was 24 ℃,with the concentrations of eugenol emulsion was 35 mg/l to dipping bath carps.The pharmacokinetic results of plasma,Cmax was 16.955μg/ml,t1/2α was 0.085 h,t1/2β was4.156 h,AUC(0-t)was 18.081 mg/L · h.Pharmacokinetic parameters showed that the eugenol atrioventricular model was two compartment model.Metabolism equation of eugenol was C=28.573e-1.236t+ 2.695e-0.226 t.5.The change of eugenol content in tissues :The temperature was 24 ℃,with the concentrations of eugenol emulsion was 35 mg/l to dipping bath carps.At 48 h,eugenol of various tissues was undetectable.The period of 0.25 h to 0.5 h,drug concentration made thebiggest eliminate.The concentration in muscle was lowest,and minimize the extent of changes over time;Concentration in the brain in early was much higher than muscle.During the period of 0.25 ~ 0.5 h the concentration higher than that of liver and kidney.The rest of concentration and the change of concentration was similar to muscles;The concentrations in liver was the highest,the concentration in kidney came second.In addition to the 0.25~ 0.5 h,it was less than the brain,but the rest of concentrations were higher than in muscle and brain.In 0.5 h,concentrations of eugenol in liver,kidney,brain falled fast,but the eliminate at a slower pace later.6.With the condition of water temperature was 24℃and eugenol emulsion of optimal anesthetic concentration was 35 mg/l to dipping bath carps which were 0.75±0.1kg.According to the national standard of eugenol(GB2760-96)in meat safety amount was40-2000 mg/kg and the eugenol in organizations could not be detected after 48 h to say that carp muscle was safe to food,it had no use to set up the withdrawal time.7.With the condition of water temperature was 24℃and eugenol emulsion of optimal anesthetic concentration was 35 mg/l to dipping bath carps which were 0.75±0.1kg.When the concentration of brain was 19.371±6.878μg/g,carps began to enter anesthesia.When the concentration of the brain was 81.081±10.106μg/g,carps reached the depth of anesthesia.When the concentration of brain dropped to 61.455±4.137μg/g,carps began to enter recovery period.Concentrations of plasma was 4.516 ±1.056μg/ml,18.471±2.454μg/ml,12.657 ± 1.772 μg/ml.