Determination of Antibiotics Residues in Milk Using a QuEChERS Method Using Full Factorial Design and Liquid Chromatography-Tandem Mass Spectrometry

Andressa Grabsk,Carla Porto,Eduardo Pilau,Francilaine De Marchi,João De Souza,Rodolpho Do Prado,Geraldo Dos Santos

doi:10.21577/0103-5053.20190046

Abstract

This study evaluated different methods of extraction based on quick, easy, cheap, effective, rugged and safe (QuEChERS) method for analysis of the antibiotics ceftiofur, cloxacillin and enrofloxacin in milk using ultra-performance liquid chromatography-tandem mass spectrometry. The QuEChERS acetate method has been shown to be inefficient for extracting such antibiotics. The optimized QuEChERS method presented excellent recoveries, from 95 to 99%. A complete factorial design was used to evaluate the effects of variables of the clean-up step: anhydrous octadecylsilane (C18), primary secondary amine (PSA) and sodium acetate (NaAc). Linearity (R2) above 0.96 was achieved for all compounds. Accuracy and precision were assessed by recovery. Accuracy was 91-99%. Intraday precision with relative standard deviations (RSD) lower than 12.3% and interday precision lower than 12.4% were obtained. Limits of detection (LOD) and quantification (LOQ) were obtained between 1.4-6.8 and 1.5-8.7 µg L-1, respectively. The applicability was evaluated using 91 real milk samples.

Highlights

Antimicrobial resistance is a major global threat with human mortality rates of 10 million per year predicted by 2050.1 Increasing incidence of antimicrobial resistance is largely attributed to intensive use of antibiotics for humans and livestock production.[2]
Dispersive solid phase extraction (d-SPE) is the most relevant, in which different sorbents with different affinities are used for specific analytes.[9]
The validation procedure was performed based on the Manual de Garantia de Qualidade Analítica, of Ministério da Agricultura, Pecuária e Abastecimento (MAPA).[19]

Summary

Introduction

Antimicrobial resistance is a major global threat with human mortality rates of 10 million per year predicted by 2050.1 Increasing incidence of antimicrobial resistance is largely attributed to intensive use of antibiotics for humans and livestock production.[2]. Anastassiades and Lehotay[8] proposed an extraction method for the simultaneous analysis of pesticides in various agricultural food matrices, which was termed as QuEChERS (quick, easy, cheap, effective, rugged and safe). This method is based on the difference of affinities between reagents and analytes in a liquid-liquid extract improved by salting out effect. Dispersive solid phase extraction (d-SPE) is the most relevant, in which different sorbents with different affinities are used for specific analytes.[9] Yet, modifications of the QuEChERS acetate method, described by Lehotay et al.[10] were proposed to consider nature of the analytes and

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Results

Conclusion