Development, validation and application to real samples of a multiresidue LC‐MS/MS method for determination of β₂‐agonists and anabolic steroids in bovine hair

Abstract

β(2) -agonists are often abused in cattle breeding because of their effects on animal growth and meat properties. The use of β(2) -agonists as growth promoters is forbidden in the European Union (Council Directive 96/23/EC classifies them into group A of Annex I), due to their toxicity and carcinogenic properties, as for anabolic steroids, which are often administered in combination with β(2) -agonists, to promote the storage of proteins and increase muscle size. A unique confirmatory liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantitative detection of 13 β(2) -agonists and anabolic steroids plus the qualitative identification of other three analytes in bovine hair was developed and validated, according to Decision 2002/657/CE. Hair samples were washed with dichloromethane, digested within a NaOH solution and subjected to liquid-liquid extraction. The analysis was performed by high performance liquid chromatography coupled to a triple quadrupole mass spectrometer operating in the selected reaction monitoring mode. The absence of matrix interferents, together with good repeatability of both retention times and relative abundances of diagnostic transitions, allowed the correct identification of all analytes. The quantitative calibrations obtained from spiked blank hair samples proved linear in the range tested. CCα and CCβ ranged from 0.5 ng/g to 30 ng/g. Intralaboratory reproducibility (CV%) ranged between 5.0 and 17.7 and trueness between 96% ± 7% and 105% ± 8%. The applicability of the method to real positive samples was demonstrated for both β(2) -agonists and anabolic steroids. 17α-boldenone was found in most (70%) hair samples obtained from untreated animals, supporting the hypothesis of endogenous production of this steroid.

Sensitive, Fast, and Specific Immunoassays for Methyltestosterone Detection

“…However, MT in feed and its metabolite residues in meat can contribute to a series of adverse effects, therefore, several methods have been developed for the detection of MT residues. The most common detection methods are based on chromatography, including high performance liquid chromatography (HPLC) [ 10 , 11 ], liquid chromatography (LC) coupled with mass spectrometry (MS) [ 12 ], gas chromatography-mass spectrometry (GC-MS) [ 13 , 14 ] and LC-MS/MS [ 15 , 16 ]. These methods are both specific and sensitive, but they require tedious sample preparation, complicated spectral analyses, and trained operators, making these methods unsuitable for field detection.…”

Determination of steroids in bovine hair: Validation of a microwave‐assisted chemical derivatization method using liquid chromatography–tandem mass spectrometry and in vivo studies

“…The extraction protocol was studied using the naturally incurred samples containing ESB and NDF. Digestion using aqueous solutions composed of HCl, NaOH, 17,18 and TCEP 7 at 0.1 mol L −1 was compared to the methanolic extraction hereby cited, which proved to be more effective, as evidenced in Table S2 (Supporting Information). Indeed, basic digestion with NaOH led to a complete loss of the target analytes.…”

“…Indeed, basic digestion with NaOH led to a complete loss of the target analytes. After methanol was selected for extraction, different solvents and mixtures were also evaluated for LLE efficiency, among EtOAc, HEX/EtOAc (7:3, v/v), 16 EtOAc/TBME (1:1, v/v), 14 TBME, 17 and HEX, 19 which led to higher analyte peak areas (Table 1).…”

“…Extraction by aqueous acids and buffered solutions has also been reported, usually performed under overnight‐heated digestion 14–16 . The same applies to basic digestion with NaOH 17,18 . Clean‐up procedures for the purification of extracts before instrumental detection have also been carried out using in particular, liquid–liquid extraction (LLE) 14,19 or solid‐phase extraction 16,20 .…”

Overcoming nature’s paradox in skeletal muscle to optimise animal production

“…is 'to obtain more edible muscle meat for less money' but essentially to shift nutrient use towards carcass lean tissue deposition at the expense of adipose tissue (Johnson and Chung 2007). However, different countries around the world have enacted total bans or restricted the use of specific growthpromoting/growth-partitioning drugs because of their potential toxicity and carcinogenic properties (Leporati et al 2014). Other countries still allow their use in animal production, reflecting changes in consumer preferences and international politics (Higgins 2004;Stephany 2010;Bonny et al 2018;Farmer and Farrell 2018).…”