Determination of 44 foodborne stimulants and 6 progestogens in meat by QuEChERS and ultra-performance liquid chromatography-tandem mass spectrometry

To ensure the success of large-scale sporting events, prevent the contamination of food by prohibited substances, and evaluate the risk of foodborne stimulants and other hormones in food, it is necessary to establish a high-throughput, rapid, and accurate detection method for foodborne stimulants and other hormones. In this study, a QuEChERS method is proposed for the simultaneous determination of 44 foodborne stimulants and 6 progestogens using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The analyzed foodborne stimulants include 19 β2-agonists, 3 β-blockers, 11 anabolic agents, 8 glucocorticoids, and 3 diuretics. A meat sample was crushed and homogenized, following which the internal standard was added. Subsequently, the sample was shaken and extracted with water and an acetonitrile solution containing 0.5% acetic acid, then dehydrated and centrifuged with sodium chloride and anhydrous magnesium sulfate. The supernatant was purified by PSA, C18, neutral alumina, and anhydrous magnesium sulfate. It was then dried with nitrogen and concentrated. The concentrated extracts were separated using an ACQUITY BEH C18 column (100 mm×2.1 mm, 1.7 μm) with gradient elution using 0.1% formic acid-5 mmol/L ammonium acetate solution and methanol as mobile phases. The target compounds were detected by ultra-performance liquid chromatography-tandem mass spectrometry with electrospray ionization and positive ion scanning (ESI+) in the multiple reaction monitoring (MRM) mode, and quantified by the internal standard method. The linear ranges of β2-agonists and β-blockers were 0.1-20 μg/L, the linear ranges of glucocorticoids were 0.5-200 μg/L, and those of the others were approximately 0.2-50 μg/L. The linear relationships of 50 compounds were good, with correlation coefficients >0.99 in the linear ranges, and limits of quantification (LOQs) in the range of 0.1-0.4 μg/kg. The recoveries of the 50 target compounds spiked in chicken, pork, beef, lamb samples at three levels ranged from 50.3% to 119.9%, while the relative standard deviations (RSDs, n=6) ranged from 0.42% to 15.1%. Nine meat samples (including 3 beef, 3 pork, 2 chicken, and duck samples) were tested by this method and the national standard method (GB/T 21981-2008). The t test was used for statistical analysis of the hydrocortisone and cortisone contents, and no significant difference was found between the results obtained by the two methods. The developed method was used to analyze 12 beef samples from a farm. In all, 4 compounds were detected, while the other 46 were not detected. The content ranges and detection rates of the compounds were as follows: hydrocortisone: 3.3-22.6 μg/kg, 100%; cortisone: 1.5-2.1 μg/kg, 67%; androstenedione: 0.7-1.2 μg/kg, 17%; and testosterone: 0.6-1.5 μg/kg, 42%. In conclusion, the proposed method is simple, accurate, and sensitive, and hence, is suitable for the detection of foodborne stimulants and progestogens in different kinds of raw meat.