An evaluation of extraction techniques for arsenic in staple diets (fish and rice) utilising both classical and enzymatic extraction methods

Enzymatic extraction methods were evaluated with classical extraction approaches for the determination of arsenic in food. The extraction efficiency for total arsenic was determined by analysing CRM materials DORM-3 fish protein, NIES 106 rice flour and GBW10015 spinach. These were compared with total arsenic concentration determined using microwave-assisted acid digestion and ICP-MS. The total arsenic concentrations in the CRM materials were in good agreement with the certified values. Enzymatic hydrolysis using trypsin has been successfully employed to extract arsenic species in DORM-3 and fish samples. Whilst this method of hydrolysing the proteins worked well for the fish samples, an alternative approach was required to facilitate the digestion of cellulose in plant materials. However, enzymatic extraction using cellulase was found to give unsatisfactory results for both the NIES and GBW10015 CRM materials. Dilute nitric acid (1% HNO₃) was found to give a more efficient extraction for arsenic species in the same CRM materials and rice samples. The study was extended to evaluate a range of real samples. Total arsenic concentrations in 13 different types of fish tissue were determined following microwave-assisted acid digestion using nitric acid/hydrogen peroxide, followed by measurement using HPLC-ICP-MS for speciation analysis. The results obtained for fish were in the range of 3.53–98.80 µg g^?1 As (dry weight). Similarly, the results of 17 rice samples were in the range of 0.054–0.823 µg g^?1. This study demonstrates the importance of selecting an appropriate extraction technique for the quantitative measurement of arsenic species in food.     

Mercury and methylmercury bioaccessibility in swordfish

Concentrations of mercury (Hg) in swordfish (Xiphias gladius) present a food safety problem for many countries. This study analyses total Hg (t-Hg) concentrations in 27 samples of swordfish marketed in Spain in 2005 and in their bioaccessible fractions (soluble concentration in gastrointestinal medium), obtained after applying an in vitro digestion method. Methylmercury (MeHg) was also determined in the bioaccessible fractions. t-Hg concentrations in the samples were 0.41–2.11 mg kg^?1 wet weight, with a mean of 0.96 ± 0.47 mg kg^?1 wet weight. A total of 37% of the samples exceeded the Hg limit set by Spanish legislation (1.0 mg kg^?1 wet weight). Bioaccessible t-Hg concentrations were 0.17–1.72 mg kg^?1 wet weight (0.63 ± 0.4 mg kg^?1 wet weight), corresponding to 38–83% (64% ± 14%) of t-Hg. Bioaccessible MeHg concentrations, representing 94% of the bioaccessible t-Hg concentrations, were 0.16–1.53 mg kg^?1 wet weight, with a mean of 0.49 ± 0.32 mg kg^?1 wet weight. Children and adults who regularly consume this product in Spain have Hg and MeHg intakes that exceed the tolerable daily intake limits recommended by the Food and Agricultural Organization/World Health Organization (FAO/WHO) and US Environmental Protection Agency (USEPA). These results show the need for recommendations about swordfish consumption by population groups at risk in Spain.     

Natural deep eutectic solvent-based ultrasound-assisted-microextraction for extraction,pre-concentration and analysis of methylmercury and total mercury in fish and environmental waters by spectrophotometry

ABSTRACT

In this study, a simple, pH-controlled, and natural deep eutectic solvent-based microextraction (NADES-ME) procedure is proposed before the spectrophotometric analysis of methylmercury (MeHg) and total mercury in fish and environmental waters. The method is based on charge-transfer sensitive ion-pair formation between MeHg and 3-(dimethylamino)-7-(methylamino)phenothiazin-5-ium chloride (Azure B – AzB) in the presence of chelating citrate-phosphate buffer at pH 6.0, and then extraction of the formed ion-pair complex with the NADES. The important variables affecting extraction efficiency were evaluated and optimised. At optimal conditions (300 µL of 1.0 × 10^?3 mol L^?1 AzB, 600 µL NADES (betaine-sorbitol, 1:3) containing 10% water (v/v) as extractant, 375 µL acetonitrile as aprotic solvent for sonication of 7 min at power of 300 W/40 kHz, and centrifugation at 4000 rpm for 3 min, respectively), the figures of merit for MeHg include a good linearity in the concentration range of 0.7–340 µg L^?1, limits of detection and quantification of 0.25 and 0.83 µg L^?1, and pre-concentration and sensitivity enhancement factors of 120 and 95, respectively. Figures of merit for total Hg (iHg, where Hg₂²⁺ ions are predominantly present in natural oxygen-rich waters such as freshwaters and seawater as well as elemental Hg and Hg²⁺ ions at very low amounts) after pre-oxidation at pH 5.0 include linearity range of 3–270 µg L^?1 with limits of detection and quantification of 0.92 and 3.10 µg L^?1, and pre-concentration and sensitivity enhancement factors of 120 and 35, respectively. The reliability (with recovery of 92–98.7% and RSD of 1.9–5.5%) was statistically validated by analysis of two standard reference materials (SRMs) with and without spiking. The method was successfully applied to the speciation analysis of total Hg, iHg and MeHg in fish and environmental waters.     

Pesticide Residue Determination by Gas Chromatography-Tandem Mass Spectrometry as Applied to Food Safety Assessment on the Example of Some Fruiting Vegetables

A multiresidue method for the analysis of over 140 multiclass pesticides in fruiting vegetables, based on quick, easy, cheap, effective, rugged, and safe (QuEChERS) sample preparation procedure followed by gas chromatography-tandem mass spectrometry (GC-MS/MS), was established. In the validation study, the overall recoveries from spiked samples were 102?±?7, 95?±?7, and 95?±?7 % with RSD values of 7?±?3, 7?±?4, and 7?±?3 % at the spiking levels of 0.01, 0.05, and 0.5 mg kg^?1, respectively, demonstrating fitness for purpose of the method. The limit of quantification (LOQ) was 0.01 mg kg^?1 for more than 90 % of the target compounds. The analysis of over 300 samples of tomatoes, sweet peppers, and cucumbers was carried out in 2006–2014. Of these samples, 52 % contained pesticide residues but the results of the assessment of dietary exposure supported the conclusion that the presence of pesticide residues was unlikely to have a negative effect on the health of consumers. Although some of the pesticides detected in years 2006–2009 are no longer approved in the European Union member countries (namely endosulfan, oxadixyl, procymidone, propargite, and tolylfluanid), the consumer dietary exposure was low and did not exceed 12 % of the acceptable daily intake (ADI) considering both adults’ and children’s diet. Regarding short-term exposure (acute), in only one case of procymidone in sweet pepper, the acute reference dose (ARfD) for children was exceeded by 139.6 % of the ARfD.     

Determination of Menbutone in Bovine Milk and Meat Using Micellar Liquid Chromatography: Application to Injectable Dosage Forms

A simple, sensitive, and rapid method was developed for the routine identification and quantification of menbutone in different matrices by micellar liquid chromatography. Separation was performed in less than 4 min using a C₁₈ column with UV detection at 234 nm. A micellar solution composed of 0.12 M sodium dodecyl sulfate, 8 % n-butanol, and 0.3 % triethylamine in 0.02 M phosphoric acid at pH 6.0 was used as the mobile phase. The method was fully validated in accordance with International Conference on Harmonization (ICH) guidelines. The limits of detection and quantitation were 0.95 and 2.86 ng mL^?1, respectively. The method showed good repeatability, linearity, and sensitivity according to the evaluation of the validation parameters. The micellar method was successfully applied for the analysis of menbutone in its commercial injections with a mean % recovery value of 99.73?±?1.634 % and in spiked bovine milk and meat samples with a mean % recovery values in the range of 98.00–100.60 %. High extraction efficiency was obtained without matrix interference in the extraction process and in the subsequent chromatographic determination. No organic solvent was used during the pretreatment step. Hence, this method can be considered as an interesting example for green chemistry.     

Development of Multiresidue Analysis for 21 Synthetic Colorants in Meat by Microwave-Assisted Extraction–Solid-Phase Extraction–Reversed-Phase Ultrahigh Performance Liquid Chromatography

A novel ultrahigh performance liquid chromatographic method is developed for analysis of 21 synthetic colorants with different acid–base property, solubility, and polarity. The meat samples were extracted by microwave-assisted extraction followed by cleanup with solid-phase extraction. The effective separation of the colorants in meat matrixes was achieved, and no interfering peaks could be detected at the retention time of the analytes. The calibration curves showed good linearity with correlation coefficients of 0.9940–0.9999. The limits of quantification were 0.48–7.19 μg/kg. The average recovery of the 21 analytes from meat samples spiked with 25 and 75 μg?kg^?1 was 61.29–116.1 % with relative standard deviation (RSD) of <11 %. For blank beef sausage spiked with 50 μg?kg^?1 for each analyte, the intraday precision (as RSD) for 21 analytes was 1.45–9.21 % for six determinations within a day. This method has the advantages of being rapid, sensitive, accurate, and with high-throughput and can be applied for multiresidue analysis of meat samples, including six allowable azo food colorants, ten banned azo food colorants, four banned triphenylmethanes, and rhodamin B food colorant.     

液相色谱原子荧光光谱法分析测定水产动物及其制品中不同形态汞的含量

本文建立了液相色谱-原子荧光法测定水产及其制品中汞形态的检测方法,并对实验所用的流动相种类,流速,pH,进样体积,还原剂及载流(HCl)浓度进行优化,在14 min内可以实现无机汞(Hg²⁺)、甲基汞(MeHg)、乙基汞(EtHg)的分离。三种形态的汞均可以呈现良好的线性关系(r>0.9990),相对标准偏差(RSD)≤ 5.0%,以5、20、100 ng/mL这三个添加量在不同的样品基质中添加混合标准溶液进行加标回收实验,得到三种汞形态的回收率分别为80.1%~95.8%,92.1%~105%,80.3%~97.6%,实验测定检出限分别为0.021、0.014、0.018 mg/kg。用本方法检测鱼肉组织标准物质BCR463和ERM-CE464,测定值与标准值均吻合,进一步验证了本方法的准确性和可靠性。该方法前处理简单,提取效率较高,适用于实际水产及其制品中汞形态的分析测定。     

Molecularly Imprinted Nanosilica Solid-Phase Extraction for Bisphenol A in Fish Samples

This paper reports a method using molecularly imprinted nanosilica as the solid-phase extraction sorbent to extract bisphenol A (BPA) from fish samples. The selective extraction efficiency of BPA from its structure-analogous by molecularly imprinted solid-phase extraction (MISPE) was compared with commercial C18 SPE column. The reproducibility of MISPE and optimal flow rate of sample were studied. There was a linear correlation in the concentration range of 0.7–114.1 μg l^?1 of BPA (r?=?0.997). The limit of detection (LOD) based on three times ratio of signal to noise was 0.11 μg l^?1. Under optimal conditions, the proposed method was applied to the analysis of BPA in fish samples. The amount of BPA in bream was 4.00 μg kg^?1, and both concentrations of BPA in crucian and weever were below the LOD. The recoveries of BPA standard solution spiked with fish samples were in the range of 92.56–102.3 % with the relative standard deviation less than 10 %.     

Evaluation of Dispersive Liquid–Liquid Microextraction–HPLC–UV for Determination of Deoxynivalenol (DON) in Wheat Flour

A fast and simple method for the extraction of deoxynivalenol (DON) from wheat flour using dispersive liquid–liquid microextraction (DLLME) followed by high-performance liquid chromatography–UV detection has been developed and compared with immunoaffinity column cleanup (IAC) process. The influence of several important parameters on the extraction efficacy was studied. Under optimized conditions, a linear calibration curve was obtained in the range of 50–1,000 μg/L. Average recoveries of DON from spiked wheat samples at levels of 500 μg/kg for DLLME and IAC ranged from 72.9?±?1.6 and 85.5?±?3.1, respectively. A good correlation was found for spiked samples between DLLME and IAC methods. The limit of detection was 125 and 50 μg/kg for DLLME and IAC method, respectively. Advantages of DLLME method with respect to the IAC have been pointed out.     

Dehulling Characteristics of Selected Flaxseed Varieties

Dehulling characteristics of six different flaxseed varieties namely Shikha, Rashmi, Shekhar, Sweta, Padmini and Neelam at different residence times (40, 50 and 60 s) were investigated using dry dehulling. The dried flaxseed samples were dehulled in a laboratory model abrasive dehuller (rice polisher) and various dehulling parameters were determined. It was found that dehulling parameters namely embryo recovery, extraction rate, yield, hull and hullability were affected by residence time and varietal characteristics. The embryo recovery, extraction rate, yield, hull and hullability for studied varieties of flaxseed and residence time were 23.71?±?0.77–61.35?±?0.94 %, 58.26?±?2.20–79.11?±?0.43 %, 22.96?±?0.44–43.85?±?1.38 %, 16.83?±?1.75–61.54?±?5.11 % and 45.92?±?0.51–90.62?±?2.85 % (dry basis), respectively. Among the studied flaxseed varieties, Padmini and Neelam varieties were found suitable for dehulling purposes at 60 s residence time. The dehulled flaxseed (embryo), thus obtained, can be utilized for obtaining the good quality edible oil and meal for human consumption and hull as lignin-rich product.     

Application of Dispersive Liquid–Liquid Micro-extraction Using Mean Centering of Ratio Spectra Method for Trace Determination of Mercury in Food and Environmental Samples

In the present study, dispersive liquid–liquid micro-extraction has been applied for trace extraction and determination of mercury (Hg) ions in environmental samples. The mean centering of ratio spectra method was used to optimize the experimental parameters affecting the extraction of Hg. The factors influencing the extraction procedure such as type and volume of extracting and disperser solvent, concentration of chelating reagent, pH, salt effect, and centrifuge time were investigated and optimized. Under the optimized conditions, the limit of detection of the method was 0.15 μg l^?1 and enrichment factor was 39. The calibration curve was linear in the range of 0.5–100 μg l^?1 with a correlation of determination (R ²) of 0.998. The relative standard deviation for determination of 40 μg l^?1 of Hg(II) was 2.6 % (n?=?5). The proposed method was applied for the determination of Hg in pine leaf, sea and river fish, sand, and water samples as indicators of environmental pollution and cigarette with satisfactory analytical results. In comparison with other methods, the proposed method is very simple, easy, rapid, and sensitive for determination of Hg at trace levels in complex matrices.     

Monoclonal-based enzyme-linked immunosorbent assay for the detection of zearalenone in cereals

A monoclonal antibody against zearalenone (ZEA) was produced and used successfully to develop a direct competitive enzyme-linked immunosorbent assay (DC-ELISA) for the analysis of ZEA in cereals. This DC-ELISA had a limit of detection of 0.15?±?0.02 µg l^?1 and an IC₅₀ value of 1.13?±?0.16 µg l^?1. Matrix interference was minimized by dilution of the sample extract before ELISA assays. Aqueous methanol (80%) gave good extraction efficiencies, and the recovery from spiked rice, barley, and corn samples averaged between 87 and 112%. Although ZEA was detected in seven (9%) of 80 rice samples and in eight (16%) of 50 barley samples, the concentration of ZEA in samples was around or below the limit of detection of DC-ELISA. Among 38 corn samples, ZEA was detected in nine (24%) samples in the range 41.0–909.8 µg kg^?1. Re-analysis of the ELISA-positive corn samples by high-performance liquid chromatography (HPLC) confirmed that seven (18%) corn samples were positive. The ZEA results for corn showed very good agreement between DC-ELISA and a commercial AgraQant® zearalenone kit (r ²?=?0.98). Thus, the monoclonal antibody-based DC-ELISA could be applied to the preliminary screening of ZEA contamination when analysis of a large sample number is needed.     

LC-MS/MS method for the determination of the fungal pigment bikaverin in maize kernels as an indicator of ear rot

Bikaverin is a polyketide-derived pigment produced by multiple species of the fungus Fusarium, some of which can cause ear and kernel rot of maize. A method was developed for the analysis of bikaverin by high-performance liquid chromatography (LC) coupled to electrospray ionisation tandem mass spectrometry (MS/MS). The quantitative nature of the LC-MS/MS method was demonstrated over a range of concentrations of bikaverin in maize. For spike-recovery experiments utilising maize spiked with bikaverin to a level 5?µg?g^?1 of maize, the measured recovery (%) was 70.6?±?10.4. Based on the utilised method, the limit of detection (based on a signal-to-noise ratio (S/N)?>?3) was better than 0.5?µg?g^?1 from bikaverin spiked into uncontaminated ground maize. Further, the limit of quantitation (LOQ) was 3?µg?g^?1 (based on S/N?>?10) from bikaverin spiked into ground maize. The method was applied to assess contamination of maize with bikaverin following inoculation of developing maize ears with Fusarium verticillioides under agricultural field conditions.     

Enhancing Fluorescence LC Analysis of Biogenic Amines in Fish Tissues by Precolumn Derivatization with Naphthalene-2,3-dicarboxaldehyde

This work reports a sensitive high-pressure liquid chromatography method for the simultaneous determination of biogenic mono- and diamines in fish tissues. Highly fluorescent derivatives of the amines were obtained by precolumn derivatization with naphthalene-2,3-dicarboxaldehyde, in the presence of cyanide ion as the nucleophile and of heptylamine as the internal standard. The chromatographic separation was performed using a mobile phase of acetonitrile/methanol/10 % (v/v) acetone in water, delivered in gradient mode on an Inertsil ODS-3 column (250?×?4 mm i.d., 5 μm). The method was successfully applied to the analysis of fresh and canned tuna fish tissues, subjected to ultrasound-assisted liquid–liquid extraction. The variables affecting the derivatization yield as well as the extraction recovery of the analytes from the sample matrix were investigated. Results were quantified against the internal standard, according to the matrix-matched approach. Limits of detection between 2.5 and 330 mg kg^?1 sample were achieved. The precision and accuracy of assay in samples was satisfactory, yielding relative standard deviation and recovery values between 0.3 and 4.2 % and 81.0 and 102.5 %, respectively.     

Total Mercury, Inorganic Mercury and Methyl Mercury Determination in Red Wine

This study deals with the development of a method for total Hg, inorganic Hg and methylmercury (MeHg) determination in red wine by using flow injection-cold vapour generation–inductively coupled plasma mass spectrometry (FI-CVG-ICP-MS) and gas chromatography-ICP-MS (GC-ICP-MS). For Hg speciation analysis, a derivatization step was carried out using a 1% (m/v) sodium tetraphenylborate (NaBPh₄) solution, followed by extraction of Hg species and their quantification by GC-ICP-MS. The main parameters evaluated were the make-up gas flow rate, volume of the NaBPh₄ solution, time for derivatization reaction/analyte extraction and solvent used for Hg species extraction. Accuracy was evaluated by analyte recovery, whereas recoveries ranged from 99% to 104% for Hg(II) and MeHg. The limits of detection (LODs) for Hg(II) and MeHg were 0.77 and 0.80 μg L⁻¹, respectively. Wine from Argentina, Brazil, Chile and Uruguay were analysed. The wine samples were also acid digested for total Hg determination by FI-CVG-ICP-MS. The LOD of the method used for total Hg determination was 0.01 μg L⁻¹. The concentrations of Hg species in red wine measured by GC-ICP-MS were lower than the respective LODs. Only total Hg was detected in the analysed samples, where the highest concentration of Hg found was 0.55 ± 0.02 μg L⁻¹.     

A rapid liquid chromatography determination of free formaldehyde in cod

A rapid method for the determination of free formaldehyde in cod is described. It uses a simple water extraction of formaldehyde which is then derivatised with 2,4-dinitrophenylhydrazine (DNPH) to form a sensitive and specific chromophore for high-performance liquid chromatography (HPLC) detection. Although this formaldehyde derivative has been widely used in past tissue analysis, this paper describes an improved derivatisation procedure. The formation of the DNPH formaldehyde derivative has been shortened to 2 min and a stabilising buffer has been added to the derivative to increase its stability. The average recovery of free formaldehyde in spiked cod was 63% with an RSD of 15% over the range of 25–200 mg kg^?1 (n = 48). The HPLC procedure described here was also compared to a commercial qualitative procedure – a swab test for the determination of free formaldehyde in fish. Several positive samples were compared by both methods.     

The Use of Different MS Techniques to Determine Glutathione Levels in Marine Tissues

Several techniques were used, mainly mass spectrometry connected with gas chromatography, matrix-assisted laser desorption with ionization time-of-flight and mass spectrometry connected with liquid chromatography. A major problem was encountered in determining glutathione, which can be conditioned by the pH and selected reducing agents. The GSH form can be oxidized through derivatization, and a small glutathione amount in the biological samples may hinder the determination process. Another problem is the existence of a metal ion in the tested organism; therefore, often a reagent with a chelating function is added to the sample and the mobile phase in liquid chromatography is applied with appropriate polarity for GSH and GSSG. We determined the concentrations of total, reduced, and oxidized glutathione in the liver, hepatopancreas, muscle, and gonad tissues of brown shrimp (Crangon crangon) and fish (Psetta maxima and Clupea harengus membras). The highest concentrations of tGSH were recorded in the shrimp hepatopancreas (7.21?±?0.011 μmol g^?1 wet weight), in herring liver (2.85?±?0.025 μmol g^?1), and in turbot liver (1.86?±?0.063 μmol g^?1). In turn, the highest concentrations were reported for GSSG in the muscle of shrimp (0.140?±?0.000204 μmol g^?1), and in the testis of turbot (0.063?±?0.000170 μmol g^?1) and herring (0.009?±?0.000015 μmol g^?1). We also investigated seasonal changes in the concentrations of glutathione in the muscle of C. crangon shrimp in the annual cycle. The lowest values of total glutathione were recorded during spring and autumn, which could be correlated with the increase in lipid peroxidation and oxidative stress.     

Effect of Konjac Glucomannan (KGM) and Carboxymethylcellulose (CMC) on some Physico-Chemical and Mechanical Properties of Restructured Gilthead Sea Bream (Sparus aurata) Products

The development of restructured fish products and the application of new food ingredients have been used to create attractive new products and also to upgrade low-value species. The aim of this study was to evaluate the effect of konjac gum (KGM) and carboxymethylcellulose (CMC) fibres on the mechanical and physico-chemical properties of microbial transglutaminase treated restructured fish products from gilthead sea bream as well as to evaluate the effect of heat treatment and storage time. Water holding capacity, mechanical properties and colour attributes in fresh and heat-treated samples after 15 days of cold storage were measured. Results showed that these edible gums could be appropriate for making restructured products obtained from gilthead sea bream. In fresh formulations the addition of 10 g?kg^?1 of KGM or 10 g?kg^?1 of CMC presented a significant effect (p?≤?0.05) on water activity for fresh samples and the lowest value was obtained for 10 g?kg^?1 of KGM (0.975?±?0.002). Water holding capacity and adhesiveness increased due to the presence of these gums in fresh and heat-treated samples. For heat-treated samples, KGM significantly reduced (p?<?0.05) hardness, cohesiveness and chewiness. Cryo-scanning electron microscopy revealed different structures for gels containing fibres. The use of these fibres did not induce significant changes in colour parameters. Fresh or heat-treated samples stored for 15 days at 4°C showed changes in relation to the parameters investigated.     

The Mesoporous Porphyrinic Zirconium Metal-Organic Framework for Pipette-Tip Solid-Phase Extraction of Mercury from Fish Samples Followed by Cold Vapor Atomic Absorption Spectrometric Determination

In this study, the highly stable mesoporous porphyrinic zirconium metal-organic framework, namely PCN-222/MOF-545 (Zr-MOF), was prepared and used for pipette-tip solid-phase extraction of Hg(II). As a high-capacity sorbent, 4 mg of the Zr-MOF was placed into a conventional pipette tip and used, for the first time, for the fast extraction and preconcentration of mercury ions. For desorption, 50 μL of 10% HCl was used by 15 repeated aspirating/dispensing cycles, and Hg ions in elusion were measured by a cold vapor atomic absorption spectrometer. Affecting parameters on extraction efficiency were studied, and optimum conditions were established as amount of sorbent 2 mg, pH was adjusted to 5.0, the eluting volume was 15 μL, and extraction was performed on 1.8 mL of the sample. The optimal number of aspirating/dispensing cycles for extraction and desorption of analytes was found to be 10 and 15 cycles, respectively. The limit of detection of the method was found to be 20 ng L^?1 with a relative standard deviation of ≤3.1% (for seven replicate analyses of 20 μg L^?1 of mercury). Adsorption capacity and enrichment factor were 35.5 mg g^?1 and 120-fold, respectively. The proposed method was successfully applied for the determination of Hg(II) ions in fish samples.     

Simultaneous Detection of Salmonella,Listeria monocytogenes,and Staphylococcus aureus by Multiplex Real-Time PCR Assays Using High-Resolution Melting

A method combining multiplex real-time polymerase chain reaction (PCR) with high-resolution melting (HRM) analysis for rapid and specific simultaneous detection of Salmonella, Listeria monocytogenes, and Staphylococcus aureus was developed. The method included a melting-curve analysis of products and was evaluated by specificity, sensitivity and reproducibility analyses. Sensitivity and reproducibility analyses was both conducted by genomic DNA extracted from serial dilutions for each target pathogen. Assays with artificially inoculated and naturally contaminated samples after enrichment were also conducted. In the specificity test, there was no nonspecific amplification of the 44 nontarget pathogens, whereas the actual T _m values were 79.38?±?0.14, 82.54?±?0.15, and 77.36?±?0.14 °C for Salmonella, L. monocytogenes, and S. aureus, respectively. The sensitivity of the method was 3.5?×?10² CFU ml^?1 for Salmonella and L. monocytogenes and 3.5?×?10³ CFU ml^?1 for S. aureus. The coefficients of variation of T _m values ranged 0.51–1.03 % for Salmonella, 1.63–2.11 % for L. monocytogenes, and 0.75–2.17 % for S. aureus in intraassay, and ranged 0.81–2.43 % for Salmonella, 1.97–2.35 % for L. monocytogenes, and 0.93–3.93 % for S. aureus in interassay. The detection limit in artificially inoculated samples (n?=?50) was 5 CFU (25 g)^?1 food for the three tested pathogens. In the naturally contaminated samples (n?=?120),Salmonella DNA was detected by HRM, sequencing, and conventional culture-based methods at a positive rate of 25.00, 25.00, and 24.17 %, respectively; the corresponding rates for L. monocytogenes were 14.17, 14.17, and 14.17 %, respectively, while those for S. aureus were 16.7, 16.7, and 16.7 %, respectively.