Rapid and Simultaneous Determination of Anabolic Andro-Genic Steroids in Livestock and Poultry Meat Using One-Step Solid-Phase Extraction Coupled with UHPLC-MS/MS.

Anabolic androgenic steroids (AASs) are usually illegally added to animal feed because they can significantly promote animal growth and increase carcasses' leanness, which threatens the safety of animal-derived foods and indirectly hazards human health. This study aimed to establish an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous detection of twelve AAS residues in livestock and poultry meat. The homogenized samples were extracted with acetonitrile containing 1% acetic acid (v/v) and purified using the one-step extraction column. After concentration using nitrogen, the residues were redissolved in acetonitrile and then quantified with an external standard method using UHPLC-MS/MS. The results showed that the above-mentioned method had a satisfactory linear correlation (R2 ≥ 0.9903) with a concentration range of 1-100 µg/L, and the limits of detection (LODs) and quantification (LOQs) were 0.03-0.33 µg/kg and 0.09-0.90 µg/kg, respectively. With the intraday and interday precision less than 15%, the average recoveries of pork, beef, lamb, and chicken, at different spiked levels, ranged from 68.3 to 93.3%, 68.0 to 99.4%, 71.6 to 109.8%, and 70.5 to 97.7%, respectively. Overall, the established method is validated, precise, and capable of the high-throughput determination of the residues of twelve AASs in livestock and poultry meat.

Activation of Mas Oncogene-Related G Protein-Coupled Receptors Inhibits Neurochemical Alterations in the Spinal Dorsal Horn and Dorsal Root Ganglia Associated with Inflammatory Pain in Rats.

Mas oncogene-related G protein-coupled receptor C (MrgC) is unequally expressed in sensory ganglia and has been shown to modulate pathologic pain. This study investigated the mechanism underlying the effect of MrgC receptors on inflammatory pain. Intrathecal administration of the selective MrgC receptor agonist bovine adrenal medulla 8-22 (BAM8-22) (30 nmol) inhibited complete Freund's adjuvant-evoked hyperalgesia. This was associated with the inhibition of protein kinase C-γ and phosphorylated extracellular signal-regulated protein kinase in the spinal cord and/or dorsal root ganglia (DRG). The complete Freund's adjuvant injection in the hindpaw induced an increase in Gq, but not Gi and Gs, protein in the spinal dorsal horn. This increase was inhibited by the intrathecal administration of BAM8-22. The exposure of DRG cultures to bradykinin (10 µM) and prostaglandin E2 (1 µM) increased the expression of calcitonin gene-related peptide (CGRP) and neuronal nitric oxide synthase in small- and medium-sized neurons as well as the levels of CGRP, aspartate, and glutamate in the cultured medium. The bradykinin/prostaglandin E2-induced alterations were absent in the presence of BAM8-22 (10 nM). These results suggest that the activation of MrgC receptors can modulate the increase in the expression of CGRP and neuronal nitric oxide synthase as well as the release of CGRP and excitatory amino acids in DRG associated with inflammatory pain. This modulation results in the inhibition of pain hypersensitivity by suppressing the expression of Gq protein and protein kinase C-γ and extracellular signal-regulated protein kinase signaling pathways in the spinal cord and/or DRG. The present study suggests that MrgC receptors may be a novel target for relieving inflammatory pain.

Rapid and High-Throughput Determination of Sixteen ß-agonists in Livestock Meat Using One-Step Solid-Phase Extraction Coupled with UHPLC-MS/MS.

ß-agonists are illegally added to animal feed because they can greatly increase carcasses' leanness, which impairs the safety of animal-derived foods and indirectly endangers human health. This study aimed to develop an ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for determining sixteen ß-agonists in livestock meat. The homogenized samples were subjected to enzymatic hydrolysis using ß-glucuronidase/sulfatase at 40 °C for 2 h, extracted with acetonitrile containing 1% acetic acid (v/v), and purified by the one-step Qvet-AG extraction column. The residue was redissolved by 0.1% aqueous formic acid/methanol (9:1, v/v) after blow-drying by nitrogen, and then determined by UHPLC-MS/MS. The results demonstrated that the well linearity was in the range of 0.1-50 µg/L with the correlation coefficient (R2) ≥0.9928, and the limits of detection (LOD) and quantification (LOQ) were 0.01-0.11 µg/kg and 0.04-0.38 µg/kg, respectively. With intraday and interday relative standard deviations (RSDs) being less than 10%, the average recoveries of pork, beef, and lamb at various spiked levels ranged from 62.62-115.93%, 61.35-106.34%, and 62.00-111.83%, respectively. In conclusion, the established method is simple, efficient, sensitive, and suitable for the simultaneous detection of several ß-agonist residues in livestock meat.