Journal of AOAC International最新文献

{"title":"Determination of Ascorbyl Palmitate and Ascorbyl Stearate in Foods by LC-MS/MS.","authors":"Lai-Sheung Choi, Sau-Fong Chan, Kin-Wai Yeung","doi":"10.1093/jaoacint/qsaf069","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf069","url":null,"abstract":"<p><strong>Background: </strong>Ascorbyl palmitate (AP) and ascorbyl stearate (AS), together called ascorbyl esters, are antioxidants that are used as food additives to prolong the shelf-life of foods by protecting against deterioration caused by oxidation.</p><p><strong>Objective: </strong>This paper aimed to develop a simple solvent extraction and LC-MS/MS method for the determination of AP and AS in a broad range of food matrices.</p><p><strong>Methods: </strong>AP and AS in foods were extracted with methanol containing 0.5% (w/v) L-ascorbic acid and analyzed by liquid chromatography-tandem mass spectrometer in negative electrospray ionization and multiple reaction monitoring (MRM) mode.</p><p><strong>Results: </strong>The method was able to quantitatively detect AP and AS in different food matrices (canned drink, dessert, frozen fish, herbs, oils and infant formula/formulated products, etc) with recoveries of AP and AS at 73-113% and 75-113% respectively and the corresponding relative standard deviations (RSDs) of 0.3-16.1% and 0.2-13.7%. The limits of quantitation were 0.23 mg/kg for AP and 0.14 mg/kg for AS. Method applicability was also demonstrated by the detection of AP in powdered infant formula purchased from local market.</p><p><strong>Conclusion: </strong>A simple solvent extraction and LC-MS/MS method was developed, validated and demonstrated for the quantitative determination of AP and AS in various foods.</p><p><strong>Highlights: </strong>The validated method based on solvent extraction with LC-MS/MS detection offered a fast and simple method for the analysis of AP and AS in a wide range of foods.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144700774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an LC-MS/MS method for analysis of levamisole in poultry and livestock products.","authors":"Ya-Chun Chou, Pei-Jie Zheng, Chih-Neng Huang, Shu-Han Chang, Ya-Min Kao, Mei-Chih Lin, Lih-Ching Chiueh, Su-Hsiang Tseng","doi":"10.1093/jaoacint/qsaf068","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf068","url":null,"abstract":"<p><strong>Background: </strong>Levamisole is an imidazothiazole anthelmintic agent widely used in poultry and livestock. In Taiwan, the Ministry of Health and Welfare has established MRLs ranging from 0.01 to 1 μg/g for eggs, milk, and various livestock and poultry tissues.</p><p><strong>Objective: </strong>To support regulatory monitoring, a chiral LC-MS/MS method with simple sample preparation was developed for determining levamisole residues in chicken muscle, porcine muscle, liver, kidney, fat, poultry eggs, and milk from food-producing animals.</p><p><strong>Method: </strong>Separation of levamisole and its enantiomer dexamisole was achieved using an Astec Cyclobond I 2000 DMP column with 100 mM ammonium acetate and acetonitrile as the mobile phase. The sample was extracted with acetonitrile/methanol (95:5, v/v) containing 1% formic acid, followed by clean-up with acetonitrile-saturated n-hexane.</p><p><strong>Results: </strong>The method demonstrated good linearity (r > 0.995, 0.5-25 ng/mL) for three quantitative methods in all tested matrices. The method achieved a LOQ of 0.005 μg/g in all matrices. While the pre-spiked tissue calibration curve provided higher recoveries (96.3%-110.1%) by compensating for both matrix effects and analyte losses during extraction, it was more labor-intensive. In contrast, the matrix-matched calibration curve and ISTD-normalized solvent calibration curve offered slightly lower recoveries (84.8%-100.4%) but showed greater practicality for routine monitoring. All calibration strategies met the accuracy and precision criteria specified in European Commission Decision 2002/657/EC and the TFDA validation guidelines. Furthermore, levamisole residues were not detected in any of the 10 commercial livestock and poultry products analyzed, confirming the applicability of the method for food surveillance.</p><p><strong>Conclusions: </strong>A simple and rapid LC-MS/MS method was developed for the determination of levamisole in poultry and livestock matrices, demonstrating satisfactory sensitivity, accuracy, and selectivity. The method is suitable for routine monitoring and large-scale surveillance of levamisole residues in food products.</p><p><strong>Highlights: </strong>Development and validation of an LC-MS/MS method involving simple solvent extraction for chiral determination of levamisole in livestock and poultry products with satisfactory sensitivity, accuracy, and selectivity.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144692844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development Of A PCR-Based Lateral Flow Strip Assay for the Rapid Detection of Dried Sika Deer Antler Products.","authors":"Xingmei Gao, Shan Jiang, Wang Pan, Liping An, Guangyu Xu, Xiao Guo, Chang Liu, Hongyu Wu, Xiao Han","doi":"10.1093/jaoacint/qsaf066","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf066","url":null,"abstract":"<p><strong>Background: </strong>Sika deer antler, a traditional and highly valued Chinese medicinal material, faces challenges in standardization and application of identification techniques due to various limitations in existing animal-derived detection methods.</p><p><strong>Objective: </strong>To develop a rapid and visual method for authenticating dried sika deer antler products by integrating polymerase chain reaction (PCR) with lateral flow biosensor (LFB) technology.</p><p><strong>Methods: </strong>Sika deer-specific primers were designed, and their specificity, sensitivity, and detection limit were verified by agarose gel electrophoresis and nucleic acid test strips.</p><p><strong>Results: </strong>Sika deer-specific primers can specifically amplify a fragment of 478 bp fragment without cross-reactivity to phylogenetically related species. The PCR-LFB method demonstrated an absolute sensitivity of 1 pg/µL for sika deer DNA, with detection limits of 0.01% for red deer (Cervus canadensis), and 1% for both reindeer (Rangifer tarandus) and New Zealand deer antler.</p><p><strong>Conclusion: </strong>With exceptional specificity and minimal instrumentation requirements, this protocol provides a reliable tool for market surveillance, quality assurance, and Traditional Chinese Medicine standardization of sika deer antler products.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144644457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A green digestion method based on deep eutectic solvent for determination of essential trace elements in blood serum samples of tuberculosis children.","authors":"Aijaz Ahmed Memon, Tasneem Gul Kazi, Mohammad Nur-E-Alam, Hassan Imran Afridi, Jameel Ahmed Baig, Khalid Hussain Thebo, Ahsan Ali Memon","doi":"10.1093/jaoacint/qsaf065","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf065","url":null,"abstract":"<p><strong>Background: </strong>Tuberculosis is spreading throughout the globe, while it is a crucial cause of death especially children in developing countries. The disturbances in the concentrations of essential trace elements are associated with impaired immunity in pulmonary tuberculosis infection.</p><p><strong>Objective: </strong>In this study, the alterations in concentrations of essential trace elements, copper (Cu), iron (Fe) and Zn (Zn) in biological sample (blood serum) were determined in pulmonary tuberculosis (PTB) children, ages 5 to 10 years, before and after six month anti-tuberculosis treatment period.</p><p><strong>Method: </strong>An environmental friendly methodology to treated the serum sample by means of a deep eutectic solvent, composed of oxalic acid and choline chloride (Ox-ChCl) at diverse molar ratio, then the contents of mixture was shaken in ultrasonic bath at different temperature range (40-85 °C). Subsequently added dilute HNO3 (0.5 M), to the contents of the tubes and centrifuged. The supernatant solution was subjected to an inductive couple emission plasma spectrophotometer. Effects of various factors on efficiency of digesting on the serum samples, to determine the Cu, Fe and Zn, including, volumes of deep eutectic solvent and its mole ratio, temperature and shaking time of ultrasonic bath were checked.</p><p><strong>Results: </strong>The resulted data indicates that the PTB patients have changed profile of all three metals in their sera and this could be more due to the active disease rather than underlying deficiencies. Compared with the non-diseased children, the levels of Fe and Zn in the serum samples of PTB affected children were considerably lower (P < 0.05), while that of Cu/Zn ratio was extensively higher (P < 0.05).</p><p><strong>Conclusion: </strong>After six month treatment the levels of Fe and Zn were enhanced about 16% and 30% respectively, while 23% Cu was decreased in serum samples of PTB children. These values were slightly lower than referent values.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144644456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Determination of Organic Impurities and Subsidiary Colors in Ext. D&C Violet No. 2 (Acid Violet 43) by Ultra-High-Performance Liquid Chromatography.","authors":"Nebebech Belai, Rachel Pandian, Clark D Ridge, Jennifer L Janovick","doi":"10.1093/jaoacint/qsaf067","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf067","url":null,"abstract":"<p><strong>Background: </strong>The U.S. Food and Drug Administration (FDA) batch certifies the anthraquinone color additive Ext. D&C Violet No. 2 (XV2, \"Ext.\" stands for \"External\") to ensure that it meets requirements published in the Code of Federal Regulations (CFR). XV2 is manufactured by condensing 1,4-dihydroxy-9,10-anthracenedione (DHAQ) with p-toluidine (pT) followed by sulfonation of the condensation product at the ortho position of the pT group. Organic impurities include residual intermediates, DHAQ and pT, and reaction by-products, 1-hydroxy-9,10-anthracenedione (MHAQ) and p-toluidine-m-sulfonic acid (PTMS). Sulfonation of the condensation product at the meta position produces and isomeric subsidiary color (mXV2). Other subsidiary colors include a dye which is itself certifiable as D&C Green No. 5 (G5) and a sulfonated phthaloylcarbazole (AV43C).</p><p><strong>Objective: </strong>This paper describes a simple and sensitive UHPLC method for the determination of CFR-specified organic impurities in XV2 samples submitted to the FDA for batch certification.</p><p><strong>Methods: </strong>The UHPLC method uses a 1.7 mm particle size C-18 column with aqueous ammonium acetate and acetonitrile as eluants and photodiode array detection at two wavelengths. Analytes are identified by comparing their retention times and UV-visible spectra to those of reference standards.</p><p><strong>Results: </strong>Method validation demonstrates linearity, limits of detection, limits of quantitation, accuracy, and precision. Excellent regression coefficients for the calibration curves were obtained, with values >0.999. Overall accuracy was 98.2-104.3% and precision was 0.0075--5.27% for all analytes.</p><p><strong>Conclusion: </strong>The UHPLC method satisfies the accuracy and precision requisites for routine certification.</p><p><strong>Highlights: </strong>The UHPLC method reported here can be used for the determination of CFR-specified organic impurities including intermediate starting materials, reaction by-products, and subsidiary colors in samples of XV2 submitted to the FDA for batch certification. Method's LOD is well below the CFR specification levels.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144639080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extraction and Measurement of 1,4-Dioxane in Detergents Using Headspace Microextraction Followed by Gas Chromatography.","authors":"Azizollah Nezhadali, Masoumeh Darbanian, Mohammad Reza Mohammadian","doi":"10.1093/jaoacint/qsaf027","DOIUrl":"10.1093/jaoacint/qsaf027","url":null,"abstract":"<p><strong>Background: </strong>1,4-dioxane is a hazardous by-product commonly found in sanitary detergents due to certain manufacturing processes. Accurate detection and quantification of this compound are essential for consumer safety.</p><p><strong>Objective: </strong>This study aims to develop and optimize a method for detecting and quantifying 1,4-dioxane in sanitary detergents. The approach involves electropolymerization of graphene oxide nanocomposites on stainless-steel mesh, followed by headspace microextraction (HS-ME) and analysis using gas chromatography with a flame ionization detector (GC-FID).</p><p><strong>Methods: </strong>Graphene oxide nanocomposites were electropolymerized on stainless-steel mesh to create the absorbent material. This absorbent was used in HS-ME for extracting 1,4-dioxane from sanitary detergent samples. A Plackett-Burman design (PBD) screened seven factors, including extraction time, extraction temperature, salt addition effect, stirring speed, desorption time, type of extraction solvent, and volume of extraction solvent. Based on the screening results, a central composite design (CCD) optimized the four critical factors: extraction time, extraction temperature, stirring speed, and type of extraction solvent. Quantification of 1,4-dioxane was performed using GC-FID.</p><p><strong>Results: </strong>The optimized method demonstrated a linear range of 0.5 to 200 μg/mL with a correlation coefficient (R2) of 0.9989. The limits of detection and quantification were determined as 0.15 and 0.5 μg/mL, respectively. Method accuracy, assessed through the relative recovery percentage of 1,4-dioxane, yielded values between 91.6% and 104%. Intra-laboratory reproducibility percentages ranged from 3.2 to 6.8%.</p><p><strong>Conclusions: </strong>The developed method, using electropolymerized graphene oxide nanocomposites on stainless-steel mesh for HS-ME coupled with GC-FID, provides a sensitive and accurate approach for detecting and quantifying 1,4-dioxane in sanitary detergents.</p><p><strong>Highlights: </strong>Electropolymerization of graphene oxide nanocomposites on stainless-steel mesh was successfully implemented to create an effective absorbent for HS-ME of dioxane. A systematic optimization approach, combining PBD and CCD, identified and fine-tuned critical factors influencing extraction efficiency.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"497-505"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143672118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An Ultra-High Performance Liquid Chromatography-Mass Spectrometry Method Based on Solid-Phase Extraction Developed for the Analysis of Polyphenols in Prunus Mume Sieb. et Zucc.","authors":"Weiting Wang, Xuanxuan Fan, Naidong Chen, Jingwen Hao, Manman Zhang, Zhengjun Xie","doi":"10.1093/jaoacint/qsaf028","DOIUrl":"10.1093/jaoacint/qsaf028","url":null,"abstract":"<p><strong>Background: </strong>Prunus mume Sieb. et Zucc. (P. mume) is rich in various phenolic compounds; however, there is a lack of effective quality evaluation methods.</p><p><strong>Objective: </strong>The aim of this study was to establish and validate a quality evaluation method that combines composition and activity to evaluate P. mume.</p><p><strong>Methods: </strong>Phenolic compounds were quantified using an ultra-high performance liquid chromatography-diode array detector (UPLC-DAD) and identified using quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). A solid-phase extraction (SPE) method was used for enrichment and purification. An Eclipse Plus-C18 (2.1 × 50 mm, 5 μm) column was used for separation at a flow rate of 0.25 mL/min, column temperature of 30°C, and detection wavelength of 325 nm. 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) and ferric ion-reducing antioxidant power (FRAP) assays were used to determine the antioxidant activity of P. mume. In addition, tyrosinase inhibitory activity (TIA) was verified for the first time.</p><p><strong>Results: </strong>Twelve major phenolic compounds were identified, among which taxifolin and ferulic acid were first detected in P. mume. The established UPLC-DAD method exhibited a strong linear relationship (R2 = 0.9999). The limit of detection and limit of quantitation of the standard compounds were in the ranges of 0.001-0.014 ng/g and 0.009-0.048 ng/g, respectively. The method exhibited good linearity, precision, stability, and repeatability. The phenolic compound content was in the range of 0.38-75.88 mg/g. In addition, P. mume extract showed excellent ABTS•+ radical scavenging capacity (half maximal inhibitory concentration (IC50) = 108.93 ± 9.43 μg/mL), FRAP (10.23 ± 0.11 μM), and TIA (IC50 = 4.79 ± 0.08 mg/mL).</p><p><strong>Conclusion: </strong>The established QC method based on composition and activity is dependable and can be used for QC of P. mume.</p><p><strong>Highlights: </strong>The chromatographic analysis method is reliable and can provide a reference for QC methods for the determination of ingredients and activity in flower-based foods.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"637-647"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143694884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular Characterization, Antibiotic Resistance Pattern, and Biofilm Formation of Vibrio Parahaemolyticus Isolated from Tropical Seafood.","authors":"Ezhil Nilavan, Akshara Kumar, Visnuvinayagam Sivam, Murugadas Vaiyapuri, Reshmi Koombankallil, Toms Cheriyath Joseph, Thangaraj Raja Swaminathan","doi":"10.1093/jaoacint/qsaf037","DOIUrl":"10.1093/jaoacint/qsaf037","url":null,"abstract":"<p><strong>Background: </strong>Vibrio parahaemolyticus in seafood poses a major public health concern, particularly in tropical regions.</p><p><strong>Objective: </strong>The present study aims to isolate, assess antibiotic susceptibility, and determine the biofilm-forming ability of V. parahaemolyticus strains isolated from seafood sold in Cochin, India.</p><p><strong>Methods: </strong>One hundred seafood samples were collected from retail markets in Cochin and analyzed for V. parahaemolyticus. Phenotypic identification was confirmed through biochemical assays and molecular characterization using polymerase chain reaction (PCR) targeting toxR, tdh, and trh genes. Biofilm formation was assessed using the microtiter plate-crystal violet assay, and antibiotic resistance was determined using the disc diffusion method.</p><p><strong>Results: </strong>V. parahaemolyticus was detected in 43.0% (43/100) of the total seafood analyzed. A total of 43 isolates were confirmed by the toxR gene, of which five carried the tdh gene, while none harbored the trh gene. Antimicrobial susceptibility testing revealed 100% resistance to ampicillin, whereas all isolates were fully susceptible to chloramphenicol. The multiple antibiotic resistance (MAR) index ranged from 0.13 to 0.50. Notably, some multidrug-resistant isolates exhibited strong biofilm formation at 37°C.</p><p><strong>Conclusion: </strong>The high prevalence of antibiotic-resistant V. parahaemolyticus in seafood sold in Cochin and their ability to form biofilms underscores the need for rigorous monitoring and effective control strategies to safeguard public health.</p><p><strong>Highlights: </strong>The overall prevalence of V. parahaemolyticus in seafood from the retail market was 43.0%. The tdh gene was detected in five isolates, and none had the trh gene. All isolates exhibited resistance to ampicillin and were fully susceptible to chloramphenicol. The MAR index ranged from 0.13 to 0.50.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"612-619"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144035725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a LC-MS/MS Method for the Determination of Lauric Arginate Ethyl Ester (E243) in Food.","authors":"Yiu-Tung Wong, Ka-Lok Chan, Odi Hang-Wah Yiu, Chun-Ching Chen, Ting-Wai Leung, Kin-Wai Yeung","doi":"10.1093/jaoacint/qsaf053","DOIUrl":"10.1093/jaoacint/qsaf053","url":null,"abstract":"<p><strong>Background: </strong>Lauric arginate ethyl ester (LAE) is a natural cationic surfactant exhibiting excellent antimicrobial activity and has been approved as a safe additive in certain food according to the Codex Alimentarius Commission (CAC) General Standard for Food Additives (GSFA).</p><p><strong>Objective: </strong>This study aimed to develop an analytical method based on liquid chromatography tandem mass spectrometry (LC-MS/MS) for the analysis of LAE in food using isotope-labeled LAE as internal standard.</p><p><strong>Method: </strong>LAE in food was extracted by a mixture of acetonitrile (ACN)-water (9:1) and analyzed by LC-MS/MS via an internal standard calibration method using LAE-d23 hydrochloride as the internal standard.</p><p><strong>Results: </strong>The limit of detection values of LAE were 0.32 mg/kg in liquid samples and 1.0 mg/kg in solid samples. The limit of quantitation values were 4.4 mg/kg and 14 mg/kg in liquid and solid samples, respectively. Spike recoveries consistently fell within the range of 90 to 110% at three different fortification levels, accompanied by a RSD below 10% in each instance.</p><p><strong>Conclusions: </strong>An analytical method with LC-MS/MS detection for the analysis of LAE in various food matrixes was successfully developed, validated, and demonstrated to be fast, robust, and reliable.</p><p><strong>Highlights: </strong>The developed analytical method with LC-MS/MS detection offered a fast and efficient way to analyze LAE in various food samples during a routine food surveillance program.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"558-565"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144082955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual-Gene Based Recombinase-Aided Amplification (RAA) - Lateral Flow Strip for the Sensitive On-Site Detection of Yersinia enterocolitica in Food Samples.","authors":"Tiantian Zhou, Jinlin Shen, Yu Feng, Norman F Neumann, Min Xu, Hangtian Ma, Yuhao Cao, Yuan Jiang, Yuqiu Zhang, Junxin Xue, Jian Li, Shuai Zhi","doi":"10.1093/jaoacint/qsaf038","DOIUrl":"10.1093/jaoacint/qsaf038","url":null,"abstract":"<p><strong>Background: </strong>Yersinia enterocolitica is one of the most common foodborne pathogens worldwide. Currently, most detection methods are primarily focused on pathogenic strains of Y. enterocolitica; while it has been frequently observed that traditionally believed \"non-pathogenic\" strains of Y. enterocolitica can also cause human disease.</p><p><strong>Objective: </strong>Herein, we have developed a novel dual-recombinase-aided amplification (RAA)-lateral flow test strip assay targeting both of a pathogenic marker (ail) and a conserved gene (foxA) for the rapid detection of both pathogenic and non-pathogen Y. enterocolitica strains in food samples.</p><p><strong>Methods: </strong>Primers and probes were designed and optimized for the detection of ail and foxA genes using RAA technology. The optimal RAA-LFS assay was then evaluated for specificity and sensitivity, and validated in both chicken samples spiked with a series of ten-fold diluted Y. enterocolitica cultures and 100 natural food samples including milk, chicken, beef, pork, and salmon.</p><p><strong>Results: </strong>The newly designed assay demonstrated 100% specificity and achieved a detection limit of 17 CFU/reaction for both targets, with the entire sample preparation and detection process completed within 25 min. Additionally, the assay showed high sensitivity in spiked chicken samples, achieving a detection limit of 1.03 × 10-1 CFU/mL for both targets. Validation with the natural food samples confirmed the robustness of the assay, as the results from this new assay were in agreement with those from the commonly used traditional techniques.</p><p><strong>Conclusions: </strong>In summary, the dual RAA-LFS assay integrates two genetic markers into a single test strip, providing a rapid, cost-effective, specific, and sensitive tool for on-site detection of pathogenic and common Y. enterocolitica strains.</p><p><strong>Highlights: </strong>A novel dual RAA-LFS assay for the rapid detection of both pathogenic and common Y. enterocolitica strains was developed.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"620-627"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144059154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}