Study On Multi-residue Screening In Fishery Input And Aquaculture Products By UPLC-Q/Orbitrap HRMS

Drug residue means the existence of the matrix of drugs or its metabolites and related contaminants in any of the edible part of an animal product.With the developing of intensive breeding,it's inevitable to use drugs for therapeutic or non-therapeutic purposes and it makes drug residues one of the most significant threat for the consumers'health.In common sense,authorized use of drugs under the constructions of corresponding regulations would not cause the residues to exceed the maximum residue limits.But as a matter of fact,in real practice some farmers or plant owners may overuse authorized drugs or even use authorized drugs in forbidden situations in pursuit of maximum profit,this might very likely to cause drug residues level to exceed the maximum residue limits.Meanwhile,ignoring of withdrawal time or even use of forbidden drugs a significant threat to the safety of aquatic products.These drug residues may enter food chain and become severe threat to consumers who are exposed to the contaminated food.Some forbidden drugs and illegal additives may even have carcinogenic,teratogenic and mutagenic effects on human.Feed plays an important role in breeding as it provides energy and nutrition,so drug residues in feed is very likely to cause residues in animal tissue.It is impossible to keep aquatic products safe if we don't keep feed safe in advance,so it's significant to keep feed safe if we want to have safe aquatic products.Many countries set clear regulations on the use of drugs including where to be used and how much is allowed,and forbidden drugs should not be detected,which means the level must be below the lowest performance level of specific instrument and method.Therefore,a rapid and high efficient screening and confirmation method of drugs and contaminants in fishery drug,feed and aquatic product is necessary to establish,with which multiple residues of drugs and contaminants could be simultaneously detected.Which is of great significance to regulate the quality and safety in aquatic products and fishery inputs.Current methods of drug residues could only simultaneously detect one or a few categories of drugs,so it's not likely to know whether or not there are other residues in the sample.It is very time consuming and reagent consuming.Due to the limitation of detecting methods,it's difficult to have a quick assessment of the residues in aquatic products not to know whether or not there are contaminants or illegal additives in fishery drug and feed.So it's hard to discover the hidden danger of residues in fishery drug,feed and aquatic products and hard to deal with fishery emergency rightly and promptly.It's significant to establish a screening method to quick assess the residues in fishery drug,feed and aquatic products,providing a way for the safety assessment and quality monitoring of commercial fishery drug,feed and aquatic products.A database containing commonly used drugs and the contaminants which are very likely to appear in fishery drug,feed and aquatic products was established.Screening methods were established through study of sample pretreatment and high performance liquid chromatography coupled to Q-Exactive high resolution mass spectrometer.Results are as below:1.A database containing 500 compounds was established with TraceFinder?EFS,version 3.3?,in which there are 430 veterinary drugs,47 pesticides and 23 contaminants.Chromatographic and mass spectrometric information of 271 compounds was obtained.2.A instrumental method of 271 compounds was established with high performance liquid chromatography coupled to Q-Exactive high resolution mass spectrometer.The analysis parameters are as below:H-ESI ionization source,spray voltage:3200 V?+?,2800 V?-?;sheath gas:8 L/min;auxiliary gas:10 L/min;sweep gas:1.5 L/min;auxiliary gas heating temperature:350?,capillary temperature:325?;scan ode:Full MS/dd-MS²?TopN?.Full MS resolution:70,000;max injection time:00 ms;scan range:150¹000;MS² resolution:17,500;trigger threshold:2×10⁵;max injection time:80 ms;TopN=2.With this instrumental method,271,269,258,250,241,227 199 and 171 compounds could be simultaneously detected at 500ng/mL,100 ng/mL,50 ng/mL,20 ng/mL,10 ng/mL,5 ng/mL,2 ng/mL and 1ng/mL,respectively.3.A screening method of 258 compounds in fish drug was established.With this method,215 compounds were detected at 0.01 mg/kg?0.01 mg/L?,16 compounds were detected at 0.05 mg/kg?0.05 mg/L?,22 compounds were detected at 0.1mg/kg?0.1 mg/L?,5 compounds were detected at 0.5 mg/kg?0.5 mg/L?.4.A screening method of 175 compounds in feed was established.With this method,75 compounds were detected at 200?g/kg,136 at 50?g/kg and 116 at 10?g/kg.Recoveries of 80%of the compounds were between 30%¹10%.The mass eviation of precursor ions was below 1.9×10^-6.5.A screening method of 250 compounds in aquatic products?grass carp,shrimp,eel nd crab?was established.When fortified at 200?g/kg,245,248,248 and 215 ompounds were identified in grass carp,shrimp,eel and crab,respectively.When ortified at 50?g/kg,236,241,241 and 196 compounds were identified in grass carp,hrimp,eel and crab,respectively.When fortified at 20?g/kg,224,237,235 and 76 compounds were identified in grass carp,shrimp,eel and crab,respectively.hen fortified at 5?g/kg,200,212,214 and 138 compounds were identified in rass carp,shrimp,eel and crab,respectively.When fortified at 1?g/kg,134,161,36 and 65 compounds were identified in grass carp,shrimp,eel and crab,espectively.6.Actual sample application was conducted with 35 fish drug samples,30 feed amples and 48 aquatic samples.Among 35 fish drug samples,residues not shown n gradient table were detected in 18 samples,which belong to quinolones,etracyclinesandsulfonamides.andamong30feedsamples,urazolidone,testosterone propionate,medroxyprogesterone and diclazuril were etected and identified in 7 samples,which are all banned.Among 48 aquatic amples 29 compounds belong to sulfonamides,tetracyclines,?-agonists and uinolones etc.12 categories were detected.