Мултирезидуална анализа на β-агонисти во биолошки матрикси со примена на LC-MS/MS метод

Abstract

β-agonists are synthetically produced substances that are used in the treatment of pulmonary obstructive diseases in humans and domestic animals. In domestic animals are also used as heart tonics and tocolytics. In addition, β-agonists can be used as growth promoters in many animal species for meat production. Application of 5-10 times higher doses than the approved therapeutic doses in animals for meat production results in significant increase of muscle mass, better utilization of food, improved carcass composition and significant reduction of body fat. After application in farm animals, residues of β-agonists are accumulated in the tissues and are a potential risk to human health. There are many documented cases where the consumption of meat and liver containing β-agonists has led to serious intoxications in humans. Therefore, with Council Directive 96/22/EC, the European Union prohibits the use of β-agonists in farm animals, whereas with Council Directive 96/23/EC, the European Union prescribes the measures for monitoring these substances in animals and products of animal origin. The aim of this study was optimization and validation of confirmatory LC-MS/MS method for determination of β-agonists in the urine, muscle and liver and monitoring the presence of these substances in the above matrices, in the period of 2014-2016. With optimization of the LC-MS/MS method the chromatographic conditions and the conditions of the mass detector were optimized. For each β-agonist with the mass detector, with ESI +, were determined the parent ions and three daughter ions. The validation of the method was carried out in accordance with the Commission Decision 2002/657/EC and the following validation parameters were established: linearity, CCα, CCβ, specificity/selectivity, accuracy and precision. . In this study a total of 525 samples were analyzed, from which 274 urine samples, 150 muscle samples and 101 liver samples. From the data obtained from the validation it can be concluded that they are in accordance with the requirements of the Commission Decision 2002/657/EC. Therefore, in this study, was optimized and validated a linear, specific, selective, accurate and precise confirmatory LC-MS/MS method for the determination of β-agonists in the urine, muscle and liver derived from animals used in human nutrition. In the analyzed samples the β-agonists residues at a concentration above the CCα were not detected, which practically means that the results indicate a zero incidence of illegal or accidental use of β-agonists and that the samples are free of these harmful substances.