Abstract

A new method for detecting low levels of growth promoters in bovine meat was developed with the following goal: easy, fast and sensitive analysis of a wide range of compounds, with reduced consumption of chemicals and disposables. Several classes of growth promoters were included, i.e. resorcylic acid lactones (RALs) and steroids, the latter including corticosteroids and gestagens. For sample treatment, 0.5 g of homogenised bovine meat was simultaneously disrupted and extracted in a bead-ruptor machine. The organic extraction solvent was further processed by solid-phase extraction (SPE) clean-up using 96-Well Oasis® HLB Plates. Six SPE washing steps were applied to remove matrix compounds after which the growth promoters were eluted and analysed using UHPLC-MS/MS. To achieve lower detection levels and to reduce LC-solvent consumption, a narrow-bore column with an internal diameter of 1 mm was used, instead of the conventional 2.1 mm. During analysis, the mass spectrometer was operated in negative and positive ionisation mode (ion switching). The newly developed method was validated according to the Commission Decision 2002/657. The results demonstrate that the method meets the criteria as established in this Commission Decision. The precision of the method for exogenous steroids varies between 85 and 115%, the CCα for the compounds ranges from 0.1–0.9 μg kg−1 and the expanded measurement uncertainty was lower than 36%. Compared to our current in-house methods with analysis times of 2 days for a maximum of 24 samples, the new method offers improved sample throughput (96 samples in less than 24 h) and lower detection limits.

Highlights

  • In Europe, the use of growth promoters in animal production is strictly forbidden (Council Directive 96, 1996; Directive C, 1996)

  • The use of HRAM opens the door to the detection of an unlimited number of compounds and it can pursue the perspective of untargeted strategies and retrospective analyses

  • For a more efficient workflow and to increase the sample throughput, solid-phase extraction using single SPE columns was converted to a 96-well plate format

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Summary

Introduction

In Europe, the use of growth promoters in animal production is strictly forbidden (Council Directive 96, 1996; Directive C, 1996). In a recent paper of Luo et al (Luo et al, 2016), the use of hollow-fibre liquid-phase microextraction (HF-LPME) was combined in a 96-wells format to extract 16 steroids from urine, milk and plasma in only a few hours. Another fast method is based on a modified Quick Easy Cheap Effective Rugged Safe (QuEChERS) approach (Attalah et al, 2016) to extract four steroids from meat. The use of HRAM opens the door to the detection of an unlimited number of compounds and it can pursue the perspective of untargeted strategies and retrospective analyses

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