Validation of a multi-residue method in UHPLC-MS/MS for determination of ten veterinary antibiotic residues in Oncorhynchus mykiss

Abstract

In Peru there are regulated protocols for monitoring antibiotic residues in the food industry. However, the aquaculture sector, in continued growth, does not have routine monitoring checks, although the use of antibiotics in aquaculture products is common. The lack of monitoring is partly due to the fact that the Peruvian regulatory body (National Fisheries Health Agency [Organismo Nacional de Sanidad Pesquera, SANIPES]) does not have validated, regulated or accepted tests to determine and quantify the presence of veterinary residues in aquaculture products. Therefore, to identify and determine if the residues are within the maximum limit allowed by national regulations, we aimed to validate a multiresidue method of 10 antibiotics through the ultra-high performance liquid chromatography coupled to a triple quadrupole mass spectrometric (UHPLC-MS/MS), assessed in rainbow trout that is the aquaculture product with the highest national production. The validation includes parameters of selectivity, linearity, sensitivity, detection limit, quantification limit, recovery range and precision, based on the acceptability criteria of the chemical method validation guide of the Food and Veterinary Medicine Program of the Food and Drug Administration of the United States (FDA). After the validation of the method, the levels of antibiotic residues in rainbow trout samples acquired at random in 6 different markets in North Lima (Los Olivos, Independencia, Comas, Puente Piedra, San Martin) were analyzed. The method showed an adequate selectivity, lower than 13.82%; a linearity with values ​​greater than 0.99; a sensitivity with a vertical trend for most antibiotics; detection and quantification limits lower than 1,994 µg/kg and 6,647 µg/kg, respectively; a recovery within the range of 90% to 115% and an accuracy of less than 9%. Traces of oxytetracycline were detected in one of the market samples. The method proved to be adequate for the determination of 10 antibiotic residues, being a useful alternative for monitoring aquaculture products.