Rapid simultaneous determination of 160 drugs in urine and blood of livestock and poultry by ultra-high-performance liquid chromatography-tandem mass spectrometry

Abstract

In order to address the specific question of food safety in livestock and poultry, it is imperative to monitor veterinary drugs at every moment in the process of livestock and poultry breeding. Thus, multi-residue analysis of a wide variety of drugs using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) has become a tool of critical significance, especially for veterinary drug monitoring programs. A total of 160 compounds, belonging to 17 different families of veterinary drugs, were investigated in the urine and blood of livestock and poultry. Drug samples were extracted using a slightly acidic acetonitrile solution. The QuEChERS (quick, easy, cheap, effective, rugged, and safe) preparation method, combined with dispersive solid phase extraction (d-SPE) was compared with the approach of solid phase extraction (SPE). In the end, the QuEChERS extraction procedure was selected to reduce matrix effects and efficiently extract target veterinary drugs, and d-SPE was applied as a cleanup step. Electrospray ionization coupled with positive dynamic multiple reaction monitoring (dMRM) was utilized for the analysis of 160 different drugs in a single chromatographic run of 24 min. The efficiency of this method was evaluated using 7 matrices (pig blood, cattle blood, sheep blood, chicken blood, pig urine, cattle urine, and sheep urine). Good linearity was obtained for the analytes in a concentration range of 1-100 ng/mL, with correlation coefficients higher than 0.990. Most of the 160 drugs studied gave estimated limits of detection (LOQs) of 1 ng/mL, with some LOQs reaching as much as 5 ng/mL. The mean recoveries at four spike-in levels of 1, 5, 10, and 50 ng/mL, ranged from 60% to 120%. The intra-day precision measurements had coefficients of variation (n = 6) <15%, and the inter-day precision measurements were below 25%. Our method was applied in real samples and proved to be adequate for routine analysis. The proposed method proved to be simple, rapid and reliable for monitoring 160 drugs in the urine and blood of livestock and poultry, and can also be used for food safety monitoring.

Keywords: Multi-residue; QuEChERS; UHPLC-MS/MS; Urine and blood; Veterinary drugs.