Poultry Muscle Research Articles

Simultaneous determination of florfenicol, its metabolite and three fluoroquinolone residues in poultry muscles via high-performance liquid chromatography with fluorescence detection

A high-performance liquid chromatography with fluorescence detection (HPLC–FLD) procedure was developed and refined to detect florfenicol (FF), its metabolite florfenicol amine (FFA), and three fluoroquinolone (FQ) residues present in poultry muscles concurrently. Sample pretreatment was conducted through liquid–liquid extraction (LLE). Ethylenediaminetetraacetic acid (EDTA), 90 % acetonitrile and acetonitrile were used as extraction solvents. The stationary phase employed was an XBridge BEH C18 column (5 μm, 4.6 mm × 150 mm), and the mobile phase consisted of a 0.01 mol/L solution of sodium dihydrogen phosphate and acetonitrile (65:35, V/V); the former contained 0.005 mol/L sodium dodecyl sulfate and 0.1 % triethylamine (pH adjusted to 4.8 with phosphoric acid). The limits of detection (LODs) and limits of quantification (LOQs) were 0.03–1.5 µg/kg and 0.1–5.0 μg/kg, respectively. The linear relationships were satisfactory within the corresponding concentration ranges, with determination coefficients (R2) ≥ 0.9997. The mean recoveries ranged from 70.34 to 94.98 %, and the intraday relative standard deviations (RSDs) and interday RSDs were 1.46–6.24 % and 1.60–7.31 %, respectively. This verified method is applicable and reliable for analyzing actual samples.

Analysis of microRNA Expression Profiles in Broiler Muscle Tissues by Feeding Different Levels of Guanidinoacetic Acid.

The aim of this study was to explore the molecular mechanisms through which different levels of GAA affect chicken muscle development by influencing miRNA expression, to lay a theoretical foundation for the identification of key functional small RNAs related to poultry muscle development, and to provide new insights into the regulatory mechanisms of GAA on muscle development and meat quality in broilers. It provides a new theoretical basis for using GAA as a feed additive to improve feed performance. Small RNA sequencing technology was utilized to obtain the expression profiles of miRNA in the broiler pectoral muscle fed with different levels of GAA (0 g/kg, 1.2 g/kg and 3.6 g/kg). An analysis of differentially expressed miRNAs revealed 90 such miRNAs in the three combination comparisons, with gga-miR-130b-5p exhibiting significant differences across all three combinations. Furthermore, three of the differentially expressed miRNAs were performed by RT-qPCR verification, yielding results consistent with those obtained from small RNA sequencing. Target gene prediction, as well as the GO and KEGG enrichment analysis of differentially expressed miRNAs, indicated their involvement in muscle cell differentiation and other processes, particularly those associated with the MAPK signaling pathway. This study has, thus, provided valuable insights and resources for the further exploration of the miRNA molecular mechanism underlying the influence of guanidine acetic acid on broiler muscle development. Combined with previous studies and small RNA sequencing, adding 1.2 g/kg GAA to the diet can better promote the muscle development of broilers.

A comprehensive evaluation of lipid profiles and nutritional quality in different animal source muscle tissues

Lipid composition is crucial for assessing the nutrition and flavor of animal-derived raw materials, a key concern for consumers. Based on high-resolution mass spectrometry and chemical pattern recognition, the intermuscular lipids fingerprint of seven types of livestock, poultry, and fish muscle were analyzed. In total, 704 lipids were detected, containing fourteen lipid molecules, with significant content differences among samples, notably triglycerides, phosphatidylcholines, phosphatidylethanolamines, diglycerides, and fatty acids. Livestock and poultry muscle had a low content of lipids with an unsaturation degree above 4 (around 10% and 25%). These unsaturated lipid molecules in fish muscle ranged from approximately 60%–75%. Rainbow trout had higher contents of triglycerides and phosphatidylcholines with chain lengths exceeding 13. Mutton had more of these lipids within the 13–18 chain length, exceeding 0.3 g/(g·protein), which may be crucial for their unique texture and flavor. The results will help the deep processing and nutritional assessment of muscle foods.

Comparative analysis of antibiotics in broiler meat using different methods

The poultry industry is one of the fastest-growing industries worldwide, and broiler chicken meat is one of the main sources of meat. To speed up the growth of chickens in a short period of time, poultry farmers use antibiotics that prevent disease and stimulate growth by increasing the rate of feed intake and reducing mortality from pathogen attacks. This study may be useful in analyzing antibiotic residues and aid in scientific research. Samples of broiler chicken meat from different producer firms were purchased from the Almaty market. The analysis was done by chromatographic method and enzyme immunoassay. The Ridascreen test kit was used for the immunoassay. This article presents the results of the study of meat of broiler chickens of domestic and foreign producers for the presence of antibiotic residues used as growth stimulants. The researches have shown that imported meat of broiler chickens from the USA has 10-20% higher antibiotic content than the permitted level by the amount of antibiotic residues, and meat from Ukraine is 10-13% higher in antibiotcs than the permitted level. The content of chloramphenicol is within the norm in all samples. Kazakhstan and Russian samples of broiler chicken meat meet the requirements of regulatory documents in force on the territory of the EAEU and the results showed that they contain only traces of antibiotics chloramphenicol and tetracycline. Analysis of imported broiler chicken meat (USA, Ukraine) showed the presence of antibiotic residues such as tetracycline and chloramphenicol, exceeding the maximum allowable level. To find out the effect of heat treatment on reducing the amount of antibiotics in broiler chicken meat, pates were made using different modes of heat treatment. As a result of heat treatment, the antibiotic content of poultry muscle tissue is significantly reduced.

Determination of ethoxyquin by ultra-high performance liquid chromatography with tandem mass spectrometry and a Singapore survey of ethoxyquin residues in eggs, egg products and poultry

In this study, an advanced ultra-high performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method was developed for quantifying ethoxyquin (EQ). The approach employed a distinctive antioxidant added extraction step designed to prevent ethoxyquin decomposition and maintain analytical precision. This method effectively determines residue levels of EQ in eggs, processed egg products, poultry muscle, salmon, and liquid milk. The method was shown to have a limit of quantitation (LOQ) for eggs, milk, salmon, and chicken muscle of 1.5 µg/kg, 1.9 µg/kg, 2.1 µg/kg, and 1.2 µg/kg, respectively. The recoveries of EQ ranged from 79.2% to 107.6%, with a relative standard deviation (RSD) below 8.4%. A surveillance study for the presence of EQ in different types of eggs and poultry muscle available in Singapore was conducted and a total of 140 samples were tested. EQ residues in all samples were found to be below the U.S. Food and Drug Administration (FDA) MRLs of 500 µg/kg. Some samples of salted and preserved eggs from China were detected with higher concentration of EQ.

Functional identification of PGM1 in the regulating development and depositing of inosine monophosphate specific for myoblasts.

Inosine monophosphate (IMP) is naturally present in poultry muscle and plays a key role in improving meat flavour. However, IMP deposition is regulated by numerous genes and complex molecular networks. In order to excavate key candidate genes that may regulate IMP synthesis, we performed proteome and metabolome analyses on the leg muscle, compared to the breast muscle control of 180-day-old Jingyuan chickens (hens), which had different IMP content. The key candidate genes identified by a differential analysis were verified to be associated with regulation of IMP-specific deposition. The results showed that the differentially expressed (DE) proteins and metabolites jointly involve 14 metabolic pathways, among which the purine metabolic pathway closely related to IMP synthesis and metabolism is enriched with four DE proteins downregulated (with higher expression in breast muscles than in leg muscles), including adenylate kinase 1 (AK1), adenosine monophosphate deaminase 1 (AMPD1), pyruvate kinase muscle isoenzyme 2 (PKM2) and phosphoglucomutase 1 (PGM1), six DE metabolites, Hypoxanthine, Guanosine, L-Glutamine, AICAR, AMP and Adenylsuccinic acid. Analysis of PGM1 gene showed that the high expression of PGM1 promoted the proliferation and differentiation of myoblasts and inhibited the apoptosis of myoblasts. ELISA tests have shown that PGM1 reduced adenosine triphosphate (ATP) and IMP and uric acid (UA), while enhancing the biosynthesis of hypoxanthine (HX). In addition, up-regulation of PGM1 inhibited the expression of purine metabolism pathway related genes, and promoted the IMP de novo and salvage synthesis pathways. This study preliminarily explored the mechanism of action of PGM1 in regulating the growth and development of myoblasts and specific IMP deposition in Jingyuan chickens, which provided certain theoretical basis for the development and utilization of excellent traits in Jingyuan chickens.

A Rapid Integrated Detection Platform for Genes Related to Duck Muscle Tissue Development

In duck breeding, the growth and development of skeletal muscle is an important factor influencing the meat production performance of ducks. Therefore, the determination of Myod and Myf5 gene expression in poultry skeletal muscle tissues can help to understand the muscle development of poultry, improve the production performance and feed conversion rate of animal organism, enhance the rapid protein deposition in animal organism, and obtain high quality and quantity of animal livestock products. In this study, a fluorescent PCR assay system for Myod and Myf5 genes was developed, and a dual integrated rapid detection platform suitable for detecting Myod and Myf5 genes in poultry muscle tissue with a sensitivity of 10 copies/μL was constructed using a set of commercial, fully automated nucleic acid analyzer with integrated detection based on magnetic bead method for nucleic acid extraction and PCR fluorescence detection. For 20 simulated samples, the integrated detection system was consistent with the results of qPCR experiments after conventional laboratory extraction, while the closed cassette-based detection reduced the chance of contamination occurrence, making the results more reliable and accurate, which is ideal for immediate on-site rapid detection.

Determination of Tilmicosin in Poultry and Pork Meat by a Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) Preparation Method and Gas Chromatography–Tandem Mass Spectrometry (GC–MS/MS)

This paper presents a novel and reliable approach for the determination of tilmicosin residues in poultry muscle and pork via gas chromatography–tandem mass spectrometry after precolumn derivatization. The samples were subjected to a quick, easy, cheap, effective, rugged, and safe preparation method, solid-phase extraction, hydrogenation, gel chromatography separation, acid hydrolysis, and derivatization with pyridine and acetic anhydride. For the blank spiked samples at the limit of quantification, 0.5 times the maximum residue limit (MRL), the MRL, and 2.0 times the MRL, the recoveries were from 76.91 to 87.99% with a precision of 2.04–5.47%. The limits of detection and quantification were 2.3–4.4 and 6.2–9.8 µg kg−1, respectively. The protocol was validated and real samples analyzed to demonstrate its practical application.

Genetic architecture and key regulatory genes of fatty acid composition in Gushi chicken breast muscle determined by GWAS and WGCNA

BackgroundFatty acids composition in poultry muscle is directly related to its tenderness, flavour, and juiciness, whereas its genetic mechanisms have not been elucidated. In this study, the genetic structure and key regulatory genes of the breast muscle fatty acid composition of local Chinese chicken, Gushi-Anka F2 resource population by integrating genome-wide association study (GWAS) and weighted gene co-expression network analysis (WGCNA) strategies. GWAS was performed based on 323,306 single nucleotide polymorphisms (SNPs) obtained by genotyping by sequencing (GBS) method and 721 chickens from the Gushi-Anka F2 resource population with highly variable fatty acid composition traits in the breast muscle. And then, according to the transcriptome data of the candidate genes that were obtained and phenotypic data of fatty acid composition traits in breast muscle of Gushi chickens at 14, 22, and 30 weeks of age, we conducted a WGCNA.ResultsA total of 128 suggestive significantly associated SNPs for 11 fatty acid composition traits were identified and mapped on chromosomes (Chr) 2, 3, 4, 5, 13, 17, 21, and 27. Of these, the two most significant SNPs were Chr13:5,100,140 (P = 4.56423e-10) and Chr13:5,100,173 (P = 4.56423e-10), which explained 5.6% of the phenotypic variation in polyunsaturated fatty acids (PUFA). In addition, six fatty acid composition traits, including C20:1, C22:6, saturated fatty acid (SFA), unsaturated fatty acids (UFA), PUFA, and average chain length (ACL), were located in the same QTL intervals on Chr13. We obtained 505 genes by scanning the linkage disequilibrium (LD) regions of all significant SNPs and performed a WGCNA based on the transcriptome data of the above 505 genes. Combining two strategies, 9 hub genes (ENO1, ADH1, ASAH1, ADH1C, PIK3CD, WISP1, AKT1, PANK3, and C1QTNF2) were finally identified, which could be the potential candidate genes regulating fatty acid composition traits in chicken breast muscle.ConclusionThe results of this study deepen our understanding of the genetic mechanisms underlying the regulation of fatty acid composition traits, which is helpful in the design of breeding strategies for the subsequent improvement of fatty acid composition in poultry muscle.

DNA Demethylation of Myogenic Genes May Contribute to Embryonic Leg Muscle Development Differences between Wuzong and Shitou Geese.

Skeletal muscle development from embryonic stages to hatching is critical for poultry muscle growth, during which DNA methylation plays a vital role. However, it is not yet clear how DNA methylation affects early embryonic muscle development between goose breeds of different body size. In this study, whole genome bisulfite sequencing (WGBS) was conducted on leg muscle tissue from Wuzong (WZE) and Shitou (STE) geese on embryonic day 15 (E15), E23, and post-hatch day 1. It was found that at E23, the embryonic leg muscle development of STE was more intense than that of WZE. A negative correlation was found between gene expression and DNA methylation around transcription start sites (TSSs), while a positive correlation was observed in the gene body near TTSs. It was also possible that earlier demethylation of myogenic genes around TSSs contributes to their earlier expression in WZE. Using pyrosequencing to analyze DNA methylation patterns of promoter regions, we also found that earlier demethylation of the MyoD1 promoter in WZE contributed to its earlier expression. This study reveals that DNA demethylation of myogenic genes may contribute to embryonic leg muscle development differences between Wuzong and Shitou geese.

Single-laboratory Validation Study and Surveillance Using an Improved Multiresidue Analytical Method for Veterinary Drugs in Livestock Products by LC-MS/MS

A method for the rapid analysis of multiclass residual veterinary drugs in poultry muscle, egg, and raw milk was validated in accordance with Japanese guidelines. Using LC-MS/MS, 20 veterinary drugs, including sulfonamides, coccidiostats, and macrolides were analyzed in one injection. Analytes were extracted from the samples with acetonitrile and then dehydrated and salted out using magnesium sulfate, trisodium citrate, and sodium chloride. This method was assessed by performing recovery tests of chicken muscle, duck muscle, egg, and raw milk spiked with 20 new target analytes at concentrations of 10 and 100 µg/kg. According to this method, 17 out of 20 target analytes satisfied the guideline criteria in chicken muscle and duck muscle, and all 20 target analytes met the criteria in egg and raw milk. The limit of quantification was less than MRLs for all analytes. Residues were detected in 4 out of 99 samples and analyzed using the validated method, finding that the levels of all residues were lower than the limits of quantification. These results suggest that continuous monitoring for a new trend of veterinary drugs is necessary.

Transcriptome analysis of the inhibitory effect of cycloleucine on myogenesis

N6-Methyladenosine (m6A) has been reported to involve and play an important role in various biological activities but seldom in poultry myogenesis. Cycloleucine usually functions as a nucleic acid methylation inhibitor, the inhibition efficiency of cycloleucine at the m6A level and corresponding dynamic changes of poultry muscle cells remain unknown. In this study, we aim to find out the effect of cycloleucine on the total N6-Methyladenosine level and its molecular mechanism for regulating myogenesis. A total of 745 differentially expressed genes (DEGs) were obtained by 10 mM, 20 mM, and 30 mM of cycloleucine treatment compared with 0 mM treatment. DEGs in 10 mM cycloleucine were significantly enriched in the biological process of skeletal muscle and satellite cell proliferation and differentiation, DEGs in 20 and 30 mM cycloleucine were enriched in some metabolic and biosynthetic processes. The trend analysis showed that 85% of all DEGs were significantly clustered into 4 files, among them 59% DEGs were dose-dependent and 26% were dose-independent, 52% DEGs were in downtrend and 33% DEGs were in uptrend. Also, the cycloleucine treatment could trigger cell cycle arrest in the G1 phase and depress myoblast cell proliferation and inhibit myotube formation. In conclusion, cycloleucine could continuously reduce the m6A level of myoblast cells, depress myoblast cell proliferation and inhibit myotube formation.

Changes in the Proteome of Poultry Muscle Tissue when Including Various Protein Supplements into Their Diet

Changes in the Proteome of Poultry Muscle Tissue when Including Various Protein Supplements into Their Diet

The Role of Incubation Conditions on the Regulation of Muscle Development and Meat Quality in Poultry.

Muscle is the most abundant edible tissue in table poultry, which serves as an important source of high protein for humans. Poultry myofiber originates in the early embryogenic stage, and the overall muscle fiber number is almost determined before hatching. Muscle development in the embryonic stage is critical to the posthatch muscle growth and final meat yield and quality. Incubation conditions including temperature, humidity, oxygen density, ventilation and lighting may substantially affect the number, shape and structure of the muscle fiber, which may produce long-lasting effect on the postnatal muscle growth and meat quality. Suboptimal incubation conditions can induce the onset of myopathies. Early exposure to suitable hatching conditions may modify the muscle histomorphology posthatch and the final muscle mass of the birds by regulating embryonic hormone levels and benefit the muscle cell activity. The elucidation of the muscle development at the embryonic stage would facilitate the modulation of poultry muscle quantity and meat quality. This review starts from the physical and biochemical characteristics of poultry myofiber formation, and brings together recent advances of incubation conditions on satellite cell migration, fiber development and transformation, and subsequent muscle myopathies and other meat quality defects. The underlying molecular and cellular mechanisms for the induced muscle growth and meat quality traits are also discussed. The future studies on the effects of external incubation conditions on the regulation of muscle cell proliferation and meat quality are suggested. This review may broaden our knowledge on the regulation of incubation conditions on poultry muscle development, and provide more informative decisions for hatchery in the selection of hatching parameter for pursuit of more large muscle size and superior meat quality.

CircMGA Depresses Myoblast Proliferation and Promotes Myotube Formation through miR-144-5p/FAP Signal

Simple SummaryCircular RNAs (circRNAs) are a class of RNAs with a circular structure that can regulate genes by acting as microRNA sponges to neutralize microRNA and release mRNA. Poultry muscle growth and development include the increase of myoblast cell numbers before birth and hypertrophy of muscle fibers after birth. In this study, we used a series of molecular and cell biology methods to identify a novel circRNA, named circMGA, that could combine miR-144-5p competitively with FAP and inhibit myoblast proliferation and promote myotube formation. The results of this study have provided new evidence and extended the knowledge about non-coding RNAs in muscle development.Circular RNAs are endogenous and abundant in skeletal muscle, and may not only be involved in regulating gene expression in a variety of ways, but also function as important regulators in poultry muscle development. Our previous research found that circMGA was differentially expressed during chicken muscle embryo development; however, as a novel circular RNA, the regulating mechanism of circMGA in myogenesis has never been studied before. In this study, we aimed to investigate the functional roles and related molecular mechanisms of circMGA in chicken primary myoblast cells. CircMGA originated from the exon 13–14 of MGA gene, was differentially expressed during embryo development and myogenesis differentiation, and could inhibit myoblast cell proliferation by repressing cell cycle related genes and promote myotube formation through MyoD and MyHC. Biotin-labeled miRNA pulldown assay and luciferase reporter assay result showed that miR-144-5p could directly target circMGA and FAP, indicating that there could be a competing endogenous RNA mechanism between circMGA and FAP. In function, miR-144-5p showed opposite regulation in myoblast cell with circMGA and FAP, just as expected. circMGA co-transfected with miR-144-5p or si-FAP could effectively eliminate the inhibition of miR-144-5p on myoblast proliferation and differentiation. In conclusion, we found a novel circRNA, named circMGA, which generated from the 13–14 exon of the MGA gene, and could inhibit myoblast proliferation and promote myotube formation by acting as the sponge of miR-144-5p and through miR-144-5p/FAP signal. Moreover, circMGA could effectively eliminate the inhibition of miR-144-5p on myoblast differentiation, thus releasing FAP and promoting myotube formation.

The effect of stocking rate and supplementary selenium on the fatty acid composition and subsequent peroxidisability of poultry muscle tissues

Selenium (Se) plays a crucial role in protecting biological materials from oxidative damage through the action of the selenoprotein glutathione peroxidase (GSH-Px), and the effectiveness of this protection is often dependent upon Se supply. Recent evidence has indicated that GSH-Px mRNA expression can be upregulated in response to potential oxidative damage risk, and that this upregulation is independent of Se supply. The current study aimed to determine the effect of Se supplementation, stocking rate and tissue fatty acid profile on GSH-Px activity in breast and thigh tissue of commercial broilers. A total of 168 Ross 308 broiler chicks were enrolled onto the study. Prior to enrolment, birds were brooded as a single group and received a starter diet containing no additional Se. The study was a 2 × 2 factorial design comprising of two levels of dietary Se (high Se, 0.5 mg/kg total Se, low Se background Se only), and two stocking rates (high, 30 kg/m2, and low, 15 kg/m2). At 15 days of age, birds were blocked by live weight and randomly allocated to one of the four treatments, with six pen replicates per treatment. At 42 days of age, one bird was randomly selected from each pen replicate, euthanased and breast and thigh tissue harvested. GSH-Px activity, thiobarbituric acid reactive substances (TBARS), and fatty acid (FA) content of these tissues were determined. There was no effect (P > 0.05) of stocking rate on GSH-Px activity or TBARS. GSH-Px activity did not differ between tissue types but was greater in high Se birds (P < 0.001) compared to low Se. TBARS concentrations were greater in thigh tissue (P < 0.001), and these thigh concentrations were greater in high Se birds (P < 0.05). There were marked differences between breast and thigh tissue in most FAs (P < 0.001), with breast generally containing greater proportions of polyunsaturated FA, so that breast tissue had a higher (P < 0.001) peroxidisability index (PI) than thigh. A positive correlation between GSH-Px activity and PI in the thigh tissue of high Se birds (Pearson Correlation 0.668; P = 0.025) may indicate that increasing susceptibility to peroxidisation in lipid-rich tissues may also upregulate GSH-Px activity in Se-replete birds. This study suggests that ensuring adequate dietary selenium could be a useful tool to mitigate adverse effects on meat quality caused by oxidation, particularly in lipid-rich meat.

Development and validation of a quantitative method for 15 antiviral drugs in poultry muscle using liquid chromatography coupled to tandem mass spectrometry

The objective of this work was to develop a quantitative multi-residue method for analysing antiviral drug residues and their metabolites in poultry meat samples. Antiviral drugs are not licensed for the treatment of influenza in food producing animals. However, there have been some reports indicating their illegal use in poultry. In this study, a method was developed for the analysis of 15 antiviral drug residues in poultry muscle (chicken, duck, quail and turkey) using liquid chromatography coupled to tandem mass spectrometry. This included 13 drugs against influenza and associated metabolites, but also two drugs employed for the treatment of herpes (acyclovir and ganciclovir). The method required the development of a novel chromatographic separation using a hydrophilic interaction chromatographic (HILIC) BEH amide column, which was necessary to retain the highly polar compounds. The analytes were detected using a triple quadrupole mass spectrometer operating in positive electrospray ionization mode. A range of different sample preparation protocols suitable for polar compounds were evaluated. The most effective procedure was based on a simple acetonitrile-based protein precipitation step followed by a further dilution in a methanol/water solution. The confirmatory method was validated according to the EU 2021/808 guidelines on different species including chicken, duck, turkey and quail. The validation was performed using various calibration curves ranging from 0.1 µg kg−1to 200 µg kg−1, according to the analyte. Depending on the analyte sensitivity, decision limits achieved ranged from 0.12 µg kg−1 for arbidol to 34.7 µg kg−1 for ribavirin. Overall, the reproducibility precision values ranged from 2.8% to 22.7% and the recoveries from 84% to 127%. The method was applied to 120 commercial poultry samples from the Irish market, which were all found to be residue-free.

Determination of Levamisole and Mebendazole and Its Two Metabolite Residues in Three Poultry Species by HPLC-MS/MS.

A high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed to simultaneously analyze levamisole (LMS) and mebendazole (MBZ) and its two metabolites, 5-hydroxymebendazole (HMBZ) and 2-amino-5-benzoylbenzimidazole (AMBZ), in poultry muscle (chicken, duck and goose). In the sample preparation process, basic ethyl acetate was used as the extraction agent, and the extracted samples were back-extracted with hydrochloric acid, purified by Oasis MCX solid-phase extraction (SPE) cartridges, and reconstituted in the initial mobile phase after being blown dry with nitrogen. Chromatographic separation was performed on an Xbridge C18 column (4.6 mm × 150 mm, 5 μm) with 0.1% formic acid in water and acetonitrile as the mobile phases, and gradient elution was performed at a flow rate of 0.6 mL/min and a column temperature of 35 °C. In blank poultry muscle samples, the spiked concentrations of LMS, MBZ, HMBZ, and AMBZ were within the range of the limit of quantitation (LOQ) to 25 μg/kg. The peak areas of the four target drugs had a good linear relationship with the concentration, and the determination coefficient (R2) values were higher than 0.9990. The average recoveries of LMS, MBZ, HMBZ, and AMBZ were 86.77–96.94%; the intraday relative standard deviations (RSDs) were 1.75–4.99% at LOQ, 0.5 maximum residue limit (MRL), 1.0 MRL, and 2.0 MRL; the interday RSDs were 2.54–5.52%; and the LODs and LOQs were 0.04–0.30 μg/kg and 0.12–0.80 μg/kg, respectively.

Risk Assessment of Nine Coccidiostats in Commercial and Home Raised Poultry.

For the first time, this paper aimed to evaluate nine ionophore and synthetic coccidiostat residues in poultry muscle samples, obtained from different production types, by solid-liquid extraction followed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The fully validated methodology was successfully applied to a total of 101 chicken and turkey samples obtained from canteens, supermarkets, and home productions in Portugal. Halofuginone, diclazuril, decoquinate, narasin, lasalocid, and salinomycin were detected in 20.8% of the samples. Home raised samples showed a greater frequency, 47.1%. The synthetic coccidiostats halofuginone, diclazuril, and decoquinate were found in averages of 0.7 μg kg-1,2.9 μg kg-1, and 3.7 μg kg-1, respectively, while averages of 1.2 μg kg-1, 1.6 μg kg-1, and 1.3 μg kg-1 were found regarding the ionophores narasin, lasalocid, and salinomycin. As for the risk assessment, values lower than 8.06% of the acceptable daily intake (ADI) were observed, indicating that exposure to coccidiostats through consumption of poultry meat does not represent risk to consumers.

Simultaneous Determination of Albendazole and Its Three Metabolites in Pig and Poultry Muscle by Ultrahigh-Performance Liquid Chromatography-Fluorescence Detection.

A fast, simple and efficient ultrahigh-performance liquid chromatography-fluorescence detection (UPLC-FLD) method for the determination of residues of albendazole (ABZ) and its three metabolites, albendazole sulfone (ABZ-SO2), albendazole sulfoxide (ABZ-SO), and albendazole-2-aminosulfone (ABZ-2NH2-SO2), in pig and poultry muscle (chicken, duck and goose) was established. The samples were extracted with ethyl acetate, and the extracts were further subjected to cleanup by utilizing a series of liquid–liquid extraction (LLE) steps. Then, extracts were purified by OASIS® PRiME hydrophilic-lipophilic balance (HLB) solid-phase extraction (SPE) cartridges (60 mg/3 mL). The target compounds were separated on an ACQUITY UPLC® BEH C18 (2.1 mm × 100 mm, 1.7 μm) chromatographic column, using a mobile phase composed of 31% acetonitrile and 69% aqueous solution (containing 0.2% formic acid and 0.05% triethylamine). The limits of detection (LODs) and limits of quantification (LOQs) of the four target compounds in pig and poultry muscle were 0.2–3.8 µg/kg and 1.0–10.9 µg/kg, respectively. The recoveries were all above 80.37% when the muscle samples were spiked with the four target compounds at the LOQ, 0.5 maximum residue limit (MRL), 1.0 MRL, and 2.0 MRL levels. The intraday relative standard deviations (RSDs) were less than 5.11%, and the interday RSDs were less than 6.29%.