Depletion Of Eugenol In The Fish Fillet Of Yellow Grouper (Epinephelus Awoara)

First,this research established a GC-MS/MS to determine eugenol residue in the fish fillet.Samples using n-hexane extraction.The extraction procedure was repeated,and the supernatant of the two extractions was then applied to the SPE column.After rinsing with 5mL hexane,the compounds were eluted by using 2 mL ethyl acetate for instrumental analysis.An Agilent 7890 N gas chromatograph(GC)coupled to a 7000 B tandem mass spectrometer(GC-MS/MS)with a triple quadrupole analyzer(QqQ)and equipped with an Agilent J&W Scientific DB-17 MS capillary column(30m � 0.25 mm,ID 0.25 ?m)was used to identify and quantify the target compounds.M/z 149 as a quantitative ion on the target,and the ion m/z 104 and m/z 77 auxiliary qualitative.Limits of detection(LODs)for eugenol,isoeugenol and methyleugenol were 0.4,1.2 and 0.2 ?g/kg,respectively.The limits of quantification(LOQs)were 1.2,4 and 0.7 ?g/kg for eugenol,isoeugenol and methyleugenol,respectively.The recoveries for eugenol and its isomers ranged from 76.4to 99.9 % with relative standard deviations(RSD)in a range from 2.18 to 15.5%.This method is quick,simple and suitable for determining the residues of eugenol,isoeugenol and methyleugenol simultaneously in batch samples of fish fillet.Second,research on temperature control without medical assistance under the survival rate of grouper,the results show that 20 ? temperature under the condition of no drugs survival rate is not high and the fish vitality is poor,cannot be returned to normal.Under25 ? with eugenol,70 mg/L high concentrations can fish into the anesthesia stage 4within 2.1 min,so that the fish into the anesthesia and recovery in 4.8 min,is suitable for the concentration in the actual transportation loading and unloading process of manual operation.8 mg/L low concentration 10 h anesthesia,the results showed that the survival rate can reach 100%,and ultimately anesthesia for anesthesia stage 2,(fish lost touch,but still maintain a balance),and is suitable for short-distance transportation process for the use of sedation.Finally,according to two different dosage anesthesia in the form of test results,determine the drug concentration in two different ways of drug residue elimination method.Grouper fish integrated by high concentration of short-time anesthesia,24 h after residues(0.148 �0.907 mg/kg)for the first time to give medicine and secondary(0.114 �0.0562mg/kg),low density long after anesthesia enrichment of fish in the highest concentration of62.3 �23.1 mg/kg,the fish in clean water recovery after 24 h residue was 39.2 �19.0mg/kg,when the cleaning time reaches 48 h content is less than 0.1 mg/kg,achieve dynamic balance.Human health risk assessment shows that high levels of repetition of anesthesia and anesthesia when low concentration long peak concentration of 62.3 mg/L is far lower than the eugenol acceptable daily intake recommended by JECFA,eugenol residue in aquatic products to human body health hazard risk is low.According to transportation and eliminate residual simulation test,the results show that high levels of repeat grouper,fish can be quickly anesthesia,after anesthesia for a long time high survival rate,and eliminate the metabolism of fast,after a certain time of storage keep safe to eat,more suitable for use in actual production process.Therefore,in order to eugenol on our country legal fish use anesthetics candidate list for reference.