Journal of AOAC International最新文献

{"title":"Per- and Polyfluoroalkyl Substances (PFAS) in Food Products by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS), AOAC Official Method 2025.07, First Action.","authors":"Toshiya Matsubara, Yusuke Mizuno, Eishi Imoto, Nozomi Maeshima, Manami Kobayashi, Okiyuki Kunihiro, William Lipps","doi":"10.1093/jaoacint/qsag014","DOIUrl":"10.1093/jaoacint/qsag014","url":null,"abstract":"<p><strong>Background: </strong>Poly- and perfluoroalkyl substances (PFAS) are fluorinated organic chemicals, which include perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS). PFAS are persistent and bioaccumulate. PFAS can enter the food supply through plants and animals grown, raised, or processed in contaminated areas.</p><p><strong>Objective: </strong>This single-laboratory validation (SLV) study was conducted for a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to determine 30 PFAS in a variety of foods, including produce, coffee, milk, protein powders, eggs, seafood, fish meat, edible offal, fish oil, baby food, and pet food, and to compare with AOAC INTERNATIONAL Standard Method Performance Requirement (SMPR®) 2023.003.</p><p><strong>Method: </strong>Matrixes were extracted using acetonitrile and QuEChERS reagents and then purified by solid-phase extraction (SPE) and concentrated under nitrogen. Fish meat did not require concentration under nitrogen, and fish oil and edible offal did not require QuEChERS reagents and sample concentration to reach the required limit of quantitation (LOQ).</p><p><strong>Results: </strong>The LC-MS/MS system provides linear responses in the range of 0.01-25 μg/kg for most target analytes, depending on the matrix. Triplicate determinations over at least three spiking concentrations were made in each matrix with quantitation by isotopic dilution. We evaluated recovery, precision, retention time, blanks, ion ratios, and signal-to-noise ratio of the qualifier ions in all matrixes to determine the LOQ. Potential interference from cholic acids were separated chromatographically.</p><p><strong>Conclusions: </strong>The method developed and validated for the analysis of PFAS in food met the LOQ and recovery and precision requirements of the SMPR.</p><p><strong>Highlights: </strong>The LC-MS/MS method developed uses a single set of operating conditions, enabling easy and rapid transfer of the method. The extraction is simplified, with only small variations necessary due to the sample matrix. The implementation of this method is cost-effective to laboratories due to minimal solvent usage and labor required to extract samples.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"695-711"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13553111/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147319186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simultaneous Analysis of Major Anthocyanins in Aronia-Containing Food Supplements by HPLC: A Greenness and Whiteness Approach.","authors":"Zeynep Büşra Abdiimamoğlu, Muhammed Zahid Kasapoglu, Gamze Ergin Kızılçay","doi":"10.1093/jaoacint/qsag026","DOIUrl":"10.1093/jaoacint/qsag026","url":null,"abstract":"<p><strong>Background: </strong>Aronia melanocarpa fruit is a valuable component of a healthy diet due to its rich antioxidant activity and high content of bioactive compounds. Recently, many food products and food supplements have emerged due to the increasing interest in aronia.</p><p><strong>Objective: </strong>In this study, a liquid chromatography method for the simultaneous quantification of the anthocyanins cyanidin-3-galactoside, cyanidin-3-arabinoside, and cyanidin-3-glucoside in aronia was developed.</p><p><strong>Methods: </strong>In this high-performance liquid chromatography (HPLC) system, the simultaneous analysis of the three active substances was performed on a Phenomenex, Gemini C18 (150 × 4.6 mm, 5 µm) column at 30°C. As a mobile phase, 1% glacial acetic acid-ACN-phosphoric acid (510:485:5, h/h/h) and 0.5% phosphoric acid solutions were used, and a gradient system was applied at a flow rate of 0.8 mL/min.</p><p><strong>Results: </strong>The developed method was validated according to ICH Q2 (R1) guidelines. Cyanidin-3-galactoside and cyanidin-3-arabinoside were found linearly in the concentration ranges of 2.5-12.5 µg/mL, and cyanidin-3-glucoside was found in the range of 0.25-4 µg/mL (r > 0.99).</p><p><strong>Conclusions: </strong>The developed and validated method was successfully applied to the analysis of samples containing aronia. Moreover, the proposed chromatographic method was assessed in terms of greenness and whiteness using the Analytical Greenness software and the White Analytical Chemistry (WAC) model.</p><p><strong>Highlights: </strong>A reliable HPLC method was developed for simultaneous quantification of three major anthocyanins in Aronia melanocarpa. The validated method was successfully applied to real aronia-containing samples.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"789-796"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13553115/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147611042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Suspension Adaptation of Immortalized Lamb Testis (iLT) Cells in Serum-Reduced Media and Evaluation of Goatpox Virus (GTPV-AV41) Replication.","authors":"Fanqi Sun, Haoyang Yu, Ting Zhang, Shaoqiang Wu, Xiangmei Lin, Caixia Wang, Zhen Yang, Chunyan Feng","doi":"10.1093/jaoacint/qsag019","DOIUrl":"10.1093/jaoacint/qsag019","url":null,"abstract":"<p><strong>Background: </strong>Goatpox is a severe viral disease in goats, and vaccination with live attenuated strains remains the most effective strategy for control. Conventional vaccine production relies on primary or Vero cells, but these substrates are limited by unstable supply, high cost, and suboptimal virus yields.</p><p><strong>Objective: </strong>This study aimed to establish a scalable suspension culture system using immortalized lamb testis (iLT) cells to support efficient replication of the goatpox virus (GTPV) AV41 strain for vaccine production.</p><p><strong>Methods: </strong>Adherent iLT cells were adapted to suspension by stepwise passaging under reduced serum. Eight commercial suspension media were screened, and four were selected for growth evaluation in 125 mL shaker flasks (20 mL working volume; 1 × 106 cells/mL), with stability assessed up to the 40th passage. For virus production, suspension cultures at 2-3 × 106 cells/mL were inoculated with GTPV-AV41 at 5-10% (v/v), samples were collected at 72-144 h post-inoculation, and viral replication was quantified by real-time quantitative PCR (RT-qPCR) using a cycle threshold (Ct) -50% tissue culture infectious dose (TCID50) calibration curve, with infection confirmed by indirect immunofluorescence.</p><p><strong>Results: </strong>Suspension-adapted iLT cells maintained robust growth for over 40 continuous passages, reaching densities exceeding 1.2 × 107 cells/mL with >95% viability. After GTPV-AV41 inoculation, the suspension culture system supported efficient viral replication, yielding a maximum viral titer of 108.00 TCID50 equivalents/mL in cell lysates at 72 h post-inoculation. Efficient viral entry and replication were further confirmed by indirect immunofluorescence assays.</p><p><strong>Conclusions: </strong>Suspension-cultured iLT cells represent a promising platform for scalable goatpox vaccine production, combining high growth performance with efficient viral replication.</p><p><strong>Highlights: </strong>This study presents a scalable suspension-culture platform by adapting iLT cells under serum-reduced conditions, enabling stable high-density growth and efficient replication of goatpox virus for cost-effective vaccine manufacturing.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"774-780"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147391911","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":"Validation and Greenness Evaluation of TLC and HPLC Methods for Analysis of Nitroxynil, Clorsulon, and Benzyl Alcohol in Flulock® Veterinary Injectable Formulation.","authors":"Nehal F Farid, Ebtehal S Bahgat, Amira M Hegazy, Rehab M Abdelfatah","doi":"10.1093/jaoacint/qsag028","DOIUrl":"10.1093/jaoacint/qsag028","url":null,"abstract":"<p><strong>Background: </strong>Nitroxynil (NIT) and clorsulon (CLO) are commonly co-formulated in veterinary injectable formulations, where benzyl alcohol is used as a preservative, necessitating selective and reliable analytical methods for their simultaneous determination.</p><p><strong>Objective: </strong>This study aimed to develop and validate sensitive, selective, and eco-friendly TLC and HPLC methods for the simultaneous determination of NIT, CLO, and benzyl alcohol in their pharmaceutical formulation.</p><p><strong>Methods: </strong>TLC separation was performed on silica gel 60 F254 plates using a mobile phase of ethyl acetate-ammonia-methanol (9.5 + 0.5 + 0.05, by volume), with densitometric detection at 254 nm. The HPLC method employed a C8 column with gradient elution using 3 mM tetrabutylammonium hydrogen sulfate (pH 6.3) and a mixture of acetonitrile and methanol, with UV detection at 215 nm. Method validation was conducted in accordance with International Conference on Harmonization (ICH) guidelines. Greenness assessment was performed using Modified Green Analytical Procedure Index (MoGAPI), Analytical Greenness Metric (AGREE), and Blue Applicability Grade Index (BAGI) tools.</p><p><strong>Results: </strong>The TLC method showed linearity over concentration ranges of 0.5-3.5 µg/band and 0.2-2 µg/band for NIT and CLO, respectively, with correlation coefficients of 0.9996 and 0.9994. The HPLC method exhibited linearity over ranges of 1-24 µg/mL and 1-21 µg/mL for the respective drugs. Both methods demonstrated high precision, accuracy, selectivity, and reproducibility, enabling effective separation of the analytes in the presence of benzyl alcohol. Greenness evaluation confirmed the low environmental impact of the proposed methods.</p><p><strong>Conclusion: </strong>The developed TLC and HPLC methods are reliable, accurate, and environmentally sustainable for the simultaneous determination of NIT, CLO, and benzyl alcohol in veterinary injectable formulations.</p><p><strong>Highlights: </strong>Simultaneous determination of NIT, CLO, and benzyl alcohol; validated TLC-densitometric and HPLC methods; successful application to Flulock® veterinary injectable formulation; high accuracy, precision, and selectivity; greenness evaluated using MoGAPI, AGREE, and BAGI tools.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"665-672"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147663391","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":"Validation of Romer Labs RapidChek® Listeria monocytogenes NextDayTM Plus Test System for the detection of Listeria monocytogenes on Select Food Matrixes and Environmental Surfaces: AOAC Performance Tested MethodSM 042501.","authors":"Juliet Wachira, Verapaz Gonzalez, Meredith Sutzko, Gregory Juck, Mark T Muldoon","doi":"10.1093/jaoacint/qsag034","DOIUrl":"10.1093/jaoacint/qsag034","url":null,"abstract":"<p><strong>Background: </strong>The RapidChek®  Listeria monocytogenes NextDay™ Plus Test System was designed to detect Listeria monocytogenes on stainless-steel and plastic environmental surfaces and in selected foods. It uses a proprietary enrichment followed by a lateral flow immunoassay to qualitatively detect L. monocytogenes.</p><p><strong>Objective: </strong>The aim of this study was to validate the Romer Labs RapidChek Listeria monocytogenes NextDay Plus Test System against the USDA-FSIS-MLG and FDA-BAM cultural reference methods for the detection of L. monocytogenes in select foods including hot dogs, frozen breaded chicken, cured ham, ice cream, and cooked shrimp, and on environmental surfaces including stainless steel and plastic (polyurethane, food grade) in an unpaired study design.</p><p><strong>Methods: </strong>The RapidChek method uses a proprietary enrichment media system, a 44 to 48 h enrichment at 30 ± 1°C, and detects L. monocytogenes on an immunochromatographic lateral flow device within 10 min. Different L. monocytogenes strains were used to spike each of the matrixes. Samples were confirmed based on the reference method confirmations and an alternate confirmation method.</p><p><strong>Results: </strong>There were 82 RapidChek presumptive positives matrixes, of which 81 were confirmed by the alternate confirmation method. The respective cultural reference methods produced 74 confirmed positives. All non-spiked samples were negative for Listeria monocytogenes by both methods. Probability of detection (POD) analysis was performed and found no statistically significant differences between methods.</p><p><strong>Conclusion: </strong>The RapidChek Listeria monocytogenes NextDay Plus Test System demonstrated performance comparable to the respective cultural reference methods while providing rapid results, supporting its use for monitoring L. monocytogenes in food and food processing environments.</p><p><strong>Highlight: </strong>The method provides the end user with a rapid and reliable tool for monitoring and control of L. monocytogenes in ready-to-eat (RTE) foods and environmental surfaces in accordance with their ongoing food safety needs.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"848-860"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147793133","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":"Validation of the RapidChek® Listeria NextDayTM Plus Test System for Detection of Listeria spp. in Matrices: Hot Dogs, Frozen Cooked Breaded Chicken, Vanilla Ice Cream, Stainless Steel, Plastic, Rubber, and Painted Concrete Surfaces: AOAC Performance Tested MethodSM 122301.","authors":"Verapaz Gonzalez, Gregory Juck, Juliet Wachira, Meredith Sutzko, Mark T Muldoon","doi":"10.1093/jaoacint/qsag032","DOIUrl":"10.1093/jaoacint/qsag032","url":null,"abstract":"<p><strong>Background: </strong>Listeria monocytogenes presents a major challenge in the food industry. Routine monitoring is essential to ensure food safety in processing environments; therefore, rapid and reliable detection of Listeria is crucial for timely and effective intervention.</p><p><strong>Objective: </strong>To validate the RapidChek®  Listeria NextDay™ Plus test system for the detection of L. monocytogenes, L. innocua, L. seeligeri, L. welshimeri, L. ivanovii, L. marthii, and L. grayi in selected foods including hot dogs, frozen cooked breaded chicken, and vanilla ice cream, and on environmental surfaces, including stainless steel, plastic, rubber, and painted concrete, using an unpaired study design.</p><p><strong>Methods: </strong>The method uses a proprietary enrichment medium, a 24 to 48 h enrichment at 30 ± 1°C and detects Listeria spp. on an immunochromatographic lateral flow device within 10 min. Listeria inoculated matrices were tested by the method, as well as the cultural reference method. The RapidChek method inclusivity was tested with 51 Listeria strains and its exclusivity using 30 non-target strains.</p><p><strong>Results: </strong>Probability of detection (POD) analysis indicated no significant differences between the candidate method and the cultural reference method in the number of positive test portions detected for hot dogs, frozen cooked breaded chicken, stainless steel, plastic, and painted concrete. For vanilla ice cream and rubber, the candidate method yielded a greater number of positive test portions, with significantly higher POD values at the 95% confidence interval. Following enrichment, the RapidChek assay detected all 51 Listeria strains evaluated and produced no false positives among the 30 non-target organisms tested.</p><p><strong>Conclusion: </strong>The candidate method performed as well as the reference method in the detection of Listeria spp. on environmental surfaces after 24 h of enrichment and in selected food matrices after 27 h of enrichment.</p><p><strong>Highlights: </strong>Enriching for 24-27 h yielded reliable presumptive results for Listeria spp.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"833-847"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147793139","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":"Analysis of Taurine in Infant Formula and Adult Nutritionals by HILIC-MS/MS, Collaborative Study, AOAC Official Method 2022.03:2026, Final Action.","authors":"Brendon D Gill, Jackie E Wood, Geert A Ten Kate","doi":"10.1093/jaoacint/qsag049","DOIUrl":"10.1093/jaoacint/qsag049","url":null,"abstract":"<p><strong>Background: </strong>Taurine is recognized as an essential growth factor and as being critical in the maintenance of functional tissue regulation. The fortification of taurine to infant formulas is necessary to provide sufficient nutrient to maintain serum levels equivalent to those of their counterparts fed on their mother's milk.</p><p><strong>Objective: </strong>To evaluate method reproducibility of AOAC First Action Official MethodSM  2022.03 for compliance with the performance requirements described in AOAC Standard Method Performance Requirements (SMPR®) 2014.013.</p><p><strong>Methods: </strong>Following protein precipitation with Carrez solutions, taurine is extracted and analyzed by hydrophilic interaction liquid chromatography-tandem mass spectrometry using multiple reaction monitoring. Stable isotope-labeled taurine internal standard is used for quantification to correct for losses in extraction and variations in ionization in the ion source.</p><p><strong>Results: </strong>Thirteen laboratories participated in the analysis of blind-duplicate samples of seven nutritional products. After outliers were removed, precision as reproducibility was found to be within limits set in SMPR 2014.013 (≤8%) for all seven test samples and all had acceptable reproducibility, with Horwitz ratio (HorRatR) values ranging from 0.4 to 0.7.</p><p><strong>Conclusion: </strong>The method described is suitable for the quantification of taurine in infant formulas and adult nutritionals. The AOAC Expert Review Panel for the Stakeholder Program for Infant Formula and Adult Nutritionals evaluated the method and accompanying validation data from this multi-laboratory testing (MLT) study in June 2025 and recommended Official Method 2022.03 for adoption as a Final Action Official Method.</p><p><strong>Highlights: </strong>An MLT study of a HILIC-MS method for the determination of taurine is described.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"828-832"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148207403","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":"Analytical Method for Detecting Herbicides, Plant Growth Regulators, and Fungicides in Agricultural Commodities: Single-Laboratory Validation.","authors":"Manisha Dhanshetty, Marco Garcia-Vaquero, Raviraj Chandrakant Shinde, Gajanan Mali, Pandit Shiragave, Sagar Atugade","doi":"10.1093/jaoacint/qsag016","DOIUrl":"10.1093/jaoacint/qsag016","url":null,"abstract":"<p><strong>Background: </strong>Pesticide residue analysis is a critical component of food safety monitoring. While multiresidue methods based on LC- or GC-MS are widely available, the analysis of highly polar or ionic pesticides remains challenging due to poor chromatographic retention, low recoveries, and repeatability.</p><p><strong>Objective: </strong>This study aimed to develop a simple, rapid, and robust analytical method for the determination of highly polar or ionic pesticides and their metabolites in selected agricultural commodities.</p><p><strong>Methods: </strong>The method targets herbicides, including glyphosate, aminomethylphosphonic acid, glufosinate, 3-methylphosphinicopropionic acid, and N-acetyl-glufosinate, the plant growth regulator ethephon, and a fungicide, fosetyl-Al, and its metabolite phosphonic acid. Sample preparation involved acidified methanolic extraction without cleanup or derivatization. Analysis was performed using LC coupled with tandem mass spectrometry (MS/MS). Chromatographic conditions were optimized to achieve baseline separation of all target analytes.</p><p><strong>Results: </strong>The method demonstrated satisfactory performance across fruit matrixes including apple, orange, tomato, cucumber, watermelon, and strawberry. Recoveries ranged from 79 to 103%, with precision values below 13% at all levels tested. The method achieved low matrix effects and met the maximum residue levels established for the target analytes in the respective commodities.</p><p><strong>Conclusion: </strong>The developed method provides a reliable and efficient approach for the analysis of highly polar pesticides and their metabolites in agricultural commodities, addressing limitations associated with conventional multiresidue methods.</p><p><strong>Highlights: </strong>An analytical method was developed for the determination of highly polar and ionic pesticides that are traditionally challenging to analyze. The method employs a simple sample preparation approach, enabling higher analytical throughput. Validation results demonstrated satisfactory performance as per the the analytical quality control requirements outlined in the SANTE/11312/2021 (V2) guidelines. The method complies with national (FSSAI) as well as international (EU) maximum residue limits.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"688-694"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147489105","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 Proline Enantiomers in Honey by FMOC-LC-FL and Storage-Associated Changes in Enantiomeric Composition.","authors":"Koichi Saito, Ayano Nakazato, Rie Ito, Hiroshi Akiyama","doi":"10.1093/jaoacint/qsag023","DOIUrl":"10.1093/jaoacint/qsag023","url":null,"abstract":"<p><strong>Background: </strong>Proline is one of the most abundant free amino acids in honey and is widely used as a quality indicator. However, its enantiomeric composition has not been systematically investigated in relation to long-term storage.</p><p><strong>Objective: </strong>This study developed a sensitive and selective method for the determination of D-proline (D-Pro) and L-proline (L-Pro) in honey using 9-fluorenylmethyl chloroformate (FMOC) derivatization followed by LC with fluorescence detection (LC-FL) and evaluated storage-associated changes in enantiomeric composition.</p><p><strong>Methods: </strong>Fresh honeys and honeys stored at ambient temperature for approximately 20 years were analyzed. After solid-phase extraction cleanup and selective masking of primary amino acids, proline enantiomers were derivatized with FMOC and quantified by LC-FL.</p><p><strong>Results: </strong>The method provided acceptable recoveries (90-98%) with a method detection limit of 0.03 μg/g, representing markedly improved sensitivity compared with a previous LC with ultraviolet detection (LC-UV) method. Both enantiomers were detected in all honeys. Long-term storage was associated with a marked decrease in L-Pro and a moderate but statistically significant increase in D-Pro (P < 0.05). Notably, the D-Pro/L-Pro ratio clearly differentiated fresh and long-term storage honeys.</p><p><strong>Conclusion: </strong>These findings indicate that long-term storage is associated with significant changes in proline enantiomer composition, likely related to non-enzymatic processes such as Maillard-type reactions. The present study introduces stereochemically resolved proline analysis as a complementary perspective for honey quality evaluation.</p><p><strong>Highlights: </strong>A sensitive FMOC-LC-FL method enabled enantiomer-resolved analysis of proline in honey, revealing storage-associated shifts in enantiomeric composition and demonstrating the potential of the D-Pro/L-Pro ratio as an additional quality indicator.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"673-679"},"PeriodicalIF":1.7,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147476604","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":"Triplex PCR Assays for Simultaneous Detection of Maize Ingredients, Maize Allergen-Encoding Genes, and the Cry1Ab Transgene in Processed Foods.","authors":"Tata Ninidze, Kakha Bitskinashvili, Tamara Kutateladze, Boris Vishnepolsky, Nelly Datukishvili","doi":"10.1093/jaoacint/qsag082","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag082","url":null,"abstract":"<p><strong>Background: </strong>Maize is widely used in processed foods but may cause allergic reactions. Reliable detection of maize ingredients, related allergens, and genetically modified organisms (GMOs) in foods is essential for food authenticity, food safety, and consumer protection. However, DNA degradation during food processing and multi-target detection remain significant analytical challenges.</p><p><strong>Objective: </strong>To develop and evaluate triplex PCR assays for the simultaneous detection of maize ingredients, allergen-encoding genes, and the Cry1Ab transgene, thereby enabling integrated food authentication, allergen and GMO screening in processed foods.</p><p><strong>Methods: </strong>Genomic DNA was extracted from certified reference materials (0-5% MON810 maize) and processed foods using the CTAB method and a commercial plant kit. Two triplex PCR systems were optimized for the detection of species-specific marker (zein), allergen-encoding genes (Zea m 8, Zea m 14), and the Cry1Ab transgene. The PCR products were analyzed via agarose gel electrophoresis.</p><p><strong>Results: </strong>The CTAB method yielded higher amounts of amplifiable DNA than the commercial kit. The developed assays enabled simultaneous detection of (i) Zea m 8, Zea m 14, and the zein gene, and (ii) Zea m 8, Zea m 14, and the Cry1Ab transgene. The assays successfully detected 0.1% MON810 reference material in the uniplex and 0.5% in the triplex systems. Analysis of processed foods confirmed reliable detection of all targets despite DNA degradation.</p><p><strong>Conclusion: </strong>The proposed triplex PCR assays offer a reliable, rapid, and cost-effective approach for the simultaneous detection of maize ingredients, allergen-encoding genes, and the Cry1Ab transgene, supporting food authentication, allergen and GMO screening in processed foods.</p><p><strong>Highlights: </strong>Triplex PCR assays enable rapid, simultaneous detection of maize ingredients, allergen-encoding genes (Zea m 8, Zea m 14, zein) and the Cry1Ab transgene in processed foods, detecting 0.1% and 0.5% MON810 reference materials by uniplex and triplex PCR, respectively.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148835683","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}