Journal of AOAC International最新文献

{"title":"Innovative Framework for Blinded Evaluation of Methods: Application to Cannabinoid Quantification in Hemp.","authors":"Andriy Tkachenko, Xiangwei Du, Jake Guag, Manisha Das, Chih-Hao Hsu, Shuping Zhang","doi":"10.1093/jaoacint/qsag039","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag039","url":null,"abstract":"<p><strong>Background: </strong>Regulatory changes and growth of hemp-derived products have increased the need for robust methods to quantify cannabinoids.</p><p><strong>Objective: </strong>To systematically evaluate a liquid chromatography photo-diode array (LC-PDA) method for 11 cannabinoids in hemp using a blinded collaborative framework, with emphasis on both fortified and endogenous analytes.</p><p><strong>Methods: </strong>A previously validated method was set up at a new‑to‑method laboratory (Veterinary Medical Diagnostic Laboratory, VMDL) and verified in-house using fortified samples. Discrepancies between certified and measured values for a hemp reference material raised questions about the method's performance for endogenous cannabinoids. To address this, an independent laboratory conducted a two-round collaborative exercise (Blinded Method Test, BMT) by preparing blind-coded samples using hemp, nettle, and hemp-nettle mixtures with various fortification levels. VMDL quantified cannabinoids by external calibration (EC) in acetonitrile (surrogate solution). Parallelism evaluation and standard-addition calibration were complementarily applied to address matrix effect and accuracy for endogenous analytes.</p><p><strong>Results: </strong>The method showed good linearity and acceptable accuracy and precision for blinded fortified hemp, nettle, and hemp-nettle samples and quality controls using EC. Parallelism results supported the use of solvent-based EC for endogenous cannabinoids in hemp. Standard-addition estimates were directionally consistent with EC but differed by about 15-45% for some analytes. These findings underscore the need for more detailed characterization of extraction efficiency for endogenous cannabinoids to strengthen accuracy assessment for incurred cannabinoids in complex botanical matrices.</p><p><strong>Conclusions: </strong>The two-round BMT demonstrates how collaborative exercises can extend conventional validation by integrating multiple matrices, parallelism testing, and complementary quantification approaches. The evaluated method is suitable for routine hemp testing at VMDL, while the findings emphasize the need to evaluate extraction efficiency for incurred versus fortified analytes in complex biomatrices.</p><p><strong>Highlights: </strong>A collaborative framework with parallelism and standard addition provides enhanced evaluation of method performance for complex botanical matrices.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147857922","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 Seven Human Milk Oligosaccharides (HMOs) in Infant Formula and Adult Nutritionals, First Action: 2025.08.","authors":"David Ellingson, Sean Austin, Thierry Bénet, Denis Cuany, Mark Damkot, Amber Wilcox","doi":"10.1093/jaoacint/qsag037","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag037","url":null,"abstract":"<p><strong>Background: </strong>The introduction of human milk oligosaccharides (HMOs) as ingredients in infant formula requires that suitable methods are available for their determination. Three methods were proposed to AOAC INTERNATIONAL for this analysis. Of the three, two methods (AOAC 2022.02 and 2022.07) were essentially based on the same approach.</p><p><strong>Objective: </strong>To conduct multi-laboratory testing (MLT) with the consolidated method from AOAC methods 2022.02 and 2022.07 for analysis of Human Milk Oligosaccharides (HMOs) in Infant Formula and Adult Nutritionals.</p><p><strong>Methods: </strong>The optimal procedural steps and settings were taken from each method to provide a method that uses ultra-high performance liquid chromatography (UHPLC) with 2-aminobenzamide (2AB) derivatization followed by hydrophilic interaction liquid chromatography (HILIC) and fluorescence detection for quantitative analysis of seven HMOs in infant formula and adult nutritionals.</p><p><strong>Results: </strong>A summary of the method parameters selected from each method along with reasoning is provided, as well as a comparison study of the previous methods. The data demonstrated that results do not differ from those of the previous methods, supporting appropriate trueness. The data also suggests the precision of the new method is likely comparable to the parent methods.</p><p><strong>Conclusion: </strong>The method was reviewed by the AOAC Expert Review Panel on Nutrients and determined that the method provided trueness to the previous methods, and additional data will be collected within the MLT to further support precision.</p><p><strong>Highlights: </strong>The method was granted AOAC First Action Official MethodsSM status.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147847860","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":"Evaluation of the Applicability of Modified QuEChERS with Small-Scale SPE for the Analysis of Neonicotinoid Insecticides in Beetroot and Paprika Powder.","authors":"Takamitsu Otake, Kaori Machida, Ayako Kimura","doi":"10.1093/jaoacint/qsaf100","DOIUrl":"10.1093/jaoacint/qsaf100","url":null,"abstract":"<p><strong>Background: </strong>As the demand for natural colorants is expected to increase in the future, it is important to analyze pesticide residues to investigate the relationship between exposure and health risks. A modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) analytical method, which uses small-scale solid-phase extraction (SPE) for the cleanup process, is increasingly used in Japan. Since this method has hardly been studied for the analysis of pesticides in natural food colorants, it is necessary to evaluate its performance.</p><p><strong>Objective: </strong>This study aimed to evaluate the applicability of a modified QuEChERS for the analysis of neonicotinoid insecticides (NEOs) in beetroot and paprika selected from natural colorants.</p><p><strong>Methods: </strong>The modified QuEChERS was evaluated by a recovery study using blank beetroot and paprika powder. The matrix effect (ME) was also evaluated in detail for the analysis by the modified QuEChERS and LC coupled with tandem mass spectrometry (LC-MS/MS). These results were compared with those obtained by the Japanese official multiresidue method.</p><p><strong>Results: </strong>The mean recovery, intraday repeatability, and interday reproducibility for the modified QuEChERS were acceptable according to the validation guideline. The MEs for the modified QuEChERS were not particularly strong for the NEOs in the beetroot and paprika even when the isotopically labeled internal standards were not used.</p><p><strong>Conclusion: </strong>The evaluation of the modified QuEChERS was successfully tested and demonstrated the suitability of this method for the analysis of NEOs in beetroot and paprika.</p><p><strong>Highlight: </strong>The modified QuEChERS is not only accurate but is also a fast and environmentally friendly method for analyzing NEOs in beetroot and paprika.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"395-405"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145338371","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":"Single-Chain Variable Fragment-Based Dot Blot, Single, and Multiple Assays for Rapid SARS-CoV-2 Diagnostics.","authors":"Dilek Çam Derin, Enes Gültekin, Irmak İçen Taşkın, Muhammed Dündar, Barış Otlu","doi":"10.1093/jaoacint/qsaf105","DOIUrl":"10.1093/jaoacint/qsaf105","url":null,"abstract":"<p><strong>Background: </strong>Antigenic detection is reliably utilized in rapid diagnostic tests and provides a significant time advantage during pandemics and epidemics. Therefore, the rapid detection of viral infections is of great importance and will remain crucial in the future. The SARS-CoV-2 outbreak, which resulted in severe losses, is the most recent example of this necessity. Among rapid diagnostic tests, lateral flow assays (LFAs) are the most practical and do not require specialized equipment, typically being developed using antibody pairs.</p><p><strong>Objective: </strong>This study aimed to recombinantly produce a single-chain variable fragment (scFv) specific to the SARS-CoV-2 spike receptor-binding domain (sRBD) and to employ it in the development of LFAs utilizing both antibody and aptamer pairs and an aptamer cocktail.</p><p><strong>Methods: </strong>Gold nanoparticles were employed as labeling agents, while both the scFv and full length forms of CR3022, along with aptamers specific to the S and N proteins, were utilized in a sandwich assay format.</p><p><strong>Results: </strong>scFv was produced at a higher concentration and biologically active. It demonstrated effective viral detection in single LFA, dot blot assay (DBA), and multiplex LFA. While single LFA successfully detected only the synthetic target, DBA and multiplex LFA selectively identified the virus in nasopharyngeal and oropharyngeal swab samples.</p><p><strong>Conclusion: </strong>Findings highlight the differences and effectiveness of using scFv in combination with other capture agents and different assay principles for the development of cost-effective and rapid diagnostic tests.</p><p><strong>Highlights: </strong>scFvs exhibit variable binding in sandwich assays depending on the combinations employed. When used in combination with an aptamer cocktail, scFvs demonstrate enhanced target binding, which is shown for the first time in this study. The use of multiple testing strategies enables a more effective viral diagnosis.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"415-424"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13148769/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145696467","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":"Validation of the QuEChERSER Method for 245 Pesticides and Environmental Contaminants in Barley and Hemp by Low-Pressure GC: Comparison of Triple Quadrupole MS/MS and Orbitrap HRMS for Qualitative and Quantitative Analysis.","authors":"Nicolás Michlig, Steven J Lehotay","doi":"10.1093/jaoacint/qsae093","DOIUrl":"10.1093/jaoacint/qsae093","url":null,"abstract":"<p><strong>Background: </strong>Monitoring laboratories are a fundamental link in the food safety chain, and regulatory demands in a competitive economy call for analytical methods that are simpler, faster, more rugged, and broader in scope. The \"quick, easy, cheap, effective, rugged, safe, efficient, and robust\" (QuEChERSER) mega-method introduced in 2021 meets these monitoring needs, which includes high sample throughput, automated cleanup of extracts, and fast low-pressure gas chromatography (LPGC).</p><p><strong>Objective: </strong>The goal of this work was to extend the QuEChERSER method to additional matrixes and more analytes using LPGC, including comparison of the analytical performances of two different mass spectrometric analyzers: triple quadrupole tandem mass spectrometry (MS/MS) and orbital ion trap (orbitrap) high-resolution (HR)MS.</p><p><strong>Methods: </strong>The QuEChERSER mega-method was validated for 245 pesticides and environmental contaminants in barley grains and hemp pellets using automated instrument top sample preparation (ITSP) coupled with LPGC-MS/MS or LPGC-HRMS (orbitrap).</p><p><strong>Results: </strong>Targeted MS/MS detection proved to be more sensitive than orbitrap using full data acquisition, leading to lower LOQs with more analytes yielding acceptable recoveries (70-120%) and repeatabilities (RSDs <20%). In barley, 89% of the compounds met validation criteria in MS/MS and 74% in HRMS, which in hemp were 81 and 66%, respectively. Qualitatively, orbitrap HRMS yielded 1% false positives compared to 3-4% in MS/MS, but due to the higher LOQs, the rates of false negatives were 14-17% in orbitrap versus 6-10% in MS/MS for the different matrixes.</p><p><strong>Conclusion: </strong>The QuEChERSER mega-method including ITSP+LPGC coupled with MS/MS or orbitrap analysis is a robust approach for multiple applications. In the comparison, MS/MS outperformed the orbitrap in terms of sensitivity, but the orbitrap advantages of easier method development, greater selectivity, and the possibility for non-targeted/retrospective analysis permit even broader expansion of analytical scope in the future.</p><p><strong>Highlights: </strong>ITSP+LPGC-MS/MS or -HRMS (orbitrap) analysis as part of the QuEChERSER mega-method is a useful and efficient way to monitor for contaminants in foods.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"281-293"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142683994","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":"TLC Method Assisted by Digital Images for Analysis of Ivermectin-Based Product.","authors":"Natália Sabina Dos Santos Galvão, Ana Carolina Kogawa","doi":"10.1093/jaoacint/qsaf087","DOIUrl":"10.1093/jaoacint/qsaf087","url":null,"abstract":"<p><strong>Background: </strong>Ivermectin (IVE) is an antiparasitic sold in the form of tablets, pastes, and injectable solutions. Neither the literature nor the official compendiums present an environmentally friendly method for analyzing the final IVE product by thin-layer chromatography (TLC) assisted by digital images. This combination strengthens the advantages of cost, handling, time, execution, and process optimization-items that green and clean analytical chemistry values and the National Environmental Methods Index (NEMI), Eco-Scale Assessment (ESA), Analytical GREEnness Metric (AGREE), and Green Analytical Procedure Index (GAPI) tools measure.</p><p><strong>Objective: </strong>The objective of this work is to develop and validate an eco-efficient, fast, economical, and easy-to-perform method for analysis of IVE injectable solution by TLC assisted by digital images.</p><p><strong>Method: </strong>Silica gel plate, microsyringe, and ethyl acetate: ethanol (13:2, v/v) as the mobile phase were used in the method. The pixels were analyzed by Image J software after the spots were photographed under UV light.</p><p><strong>Results: </strong>The method was selective when comparing standard and sample, indicative of stability by forced degradation test, linear (100-900 µg/mL), precise (RSD <2%), accurate, and rugged to modifications in the analytical process. The method was able to quantify commercial products, showing an average content of 98.95%. The greenness of the developed method presented NEMI with four green quadrants, ESA and AGREE with a score of 83 and 0.61, respectively, and GAPI predominantly green and yellow.</p><p><strong>Conclusions: </strong>The method was selective, indicative of stability, linear, precise, accurate, rugged, and green, by NEMI, ESA, AGREE, and GAPI, to quantify IVE in injectable solution. Additionally, it combined the advantages of TLC and digital image analysis.</p><p><strong>Highlights: </strong>The work shows a TLC method assisted by digital images for analysis of ivermectin-based product.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"360-365"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145093164","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":"Application of nDATA Workflow for Semi-Quantitative Screening of 1094 Pesticide Residues in Fruits and Vegetables Using UHPLC/ESI Q-Orbitrap Full MS/vDIA.","authors":"Jian Wang, Willis Chow, Jon W Wong","doi":"10.1093/jaoacint/qsaf009","DOIUrl":"10.1093/jaoacint/qsaf009","url":null,"abstract":"<p><strong>Background: </strong>Cost-effective multi-residue pesticide methods with a broad detection scope are desired for risk-based monitoring programs.</p><p><strong>Objective: </strong>The aims were to evaluate the nDATA (non-target data acquisition for target analysis) workflow using ultra high performance liquid chromatography/electrospray ionization quadrupole-Orbitrap mass spectrometry (UHPLC/ESI quadrupole-Orbitrap (Q-Orbitrap)) and semi-quantitate 1094 pesticides in fruits and vegetables.</p><p><strong>Methods: </strong>Pesticide extracts from fresh produce were prepared using the Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) procedure. nDATA was carried out by utilizing UHPLC/ESI Q-Orbitrap full MS scan and variable data independent acquisition (UHPLC/ESI Q-Orbitrap full MS/vDIA MSMS). Data were processed using a compound database (CDB, 1094 pesticides) and one-point standard calibration with internal standards for semi-quantitation. Data processing criteria were based on Retention Time (±0.5 min) and mass accuracy of a Precursor ion (±5 ppm; RTP by full MS), or Retention Time (±0.5 min) and mass accuracy of a precursor ion (±5 ppm) and that of its Fragment Ion (±10 ppm; RTFI by full MS/vDIA).</p><p><strong>Results: </strong>RTP found 1010 and 1094 pesticides, while RTFI identified 906 and 1029 pesticides at 10 and 100 μg/kg, respectively. RTF detected all 30 LC-amenable pesticides and RTFI identified 29 of 30 LC-amenable pesticides in eight proficiency testing samples. There were 42 pairs of co-eluting isomeric pesticides and 5 pairs of isobaric pesticides that were not separated by mass resolving power and/or chromatographic separation (retention time difference ΔtR <0.12 min) with the current instrument parameter settings.</p><p><strong>Conclusion: </strong>The validated nDATA workflow using UHPLC/ESI Q-Orbitrap full MS/vDIA MSMS proved to be a comprehensive detection method for semi-quantitative screening of 1094 pesticides in fruits and vegetables.</p><p><strong>Highlights: </strong>nDATA combines both non-target data acquisition and target analysis. The non-target data acquisition generates data for retrospective analysis of a large number of pesticides (over 1000). The target analysis using a CDB and a one-point standard calibration affords confidence in semi-quantitative screening results.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"326-338"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143451229","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":"Liquid- and Gas Chromatography With Tandem Mass Spectrometric Technique to Optimize and Validate the Analysis of Pesticides in Honey: A Large Scope, Multi-Class, Multi-Residue Method.","authors":"Partha P Choudhury, Sachin C Ekatpure, Abhishek Mandal, Veena Rao Udupi, Kusuma D Krishnamurthy, Nethravati Bheemiah, Seema Shenvi, Kaushik Banerjee","doi":"10.1093/jaoacint/qsaf040","DOIUrl":"10.1093/jaoacint/qsaf040","url":null,"abstract":"<p><strong>Background: </strong>Honey is a nutrient-rich food item with a complex matrix due to its diverse nutritional components. This complexity poses challenges in the analysis of pesticide residues, affecting accuracy and precision of results. A comprehensive and reliable method is required for the detection and quantification of these contaminants.</p><p><strong>Objective: </strong>To develop and validate a method for simultaneous analysis of pesticide residues in various honey types using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS).</p><p><strong>Methods: </strong>Honey samples (5 g) were extracted using acetonitrile (10 mL) and water (10 mL) for LC-MS/MS analysis, while ethyl acetate (10 mL) was used for GC-MS/MS analysis. The extract was cleaned using primary secondary amine (PSA) before pesticide residue quantification by LC-MS/MS and GC-MS/MS. The method performance was evaluated based on recoveries (70-120%) and repeatability (RSD <20%) at a limit of quantification of 0.01 mg/kg, in accordance with SANTE/11312/2021 guidelines.</p><p><strong>Results: </strong>The developed method demonstrated compliance with regulatory requirements, ensuring reliable determination of pesticide residues in honey samples. A study on matrix variability from 14 different honey samples revealed that each honey type exhibited a unique matrix effect (ME) for the targeted pesticides. To minimize this effect, the use of honey type-specific matrix-matched standards is recommended.</p><p><strong>Conclusion: </strong>The study successfully developed a simple, robust, and high-throughput analytical method for the quantitative determination of pesticide residues in honey. Given its high selectivity, sensitivity, and rugged performance, the method can be implemented in regulatory testing for the analysis of targeted compounds across a wide range of honey matrixes.</p><p><strong>Highlights: </strong>A method for pesticide residue analysis in honey was developed using LC-MS/MS and GC-MS/MS. The method showed acceptable recoveries (within 70-120%) and repeatability (RSD <20%) at an LOQ of 0.01 mg/kg. Each honey type exhibited a unique matrix effect (ME), emphasizing the need for matrix-matched standards for residue quantifications. The method demonstrated high throughput, selectivity, and sensitivity, making it suitable for regulatory testing.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"312-325"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144055855","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 and Optimization of Extraction Methodologies for the Determination of Antibiotics in Honey by UHPLC-ToF-MS.","authors":"Helena Rodrigues, Marta Leite, Beatriz Oliveira, Andreia Freitas","doi":"10.1093/jaoacint/qsaf043","DOIUrl":"10.1093/jaoacint/qsaf043","url":null,"abstract":"<p><strong>Background: </strong>The administration of antibiotics to food-producing animals, including bees, is a common practice employed to prevent or treat bacterial diseases. Such practices may result in the presence of veterinary residues in honey, potentially compromising the safety and health of consumers. In the European Union, maximum residue limits (MRLs) have not been established for pharmacological substances in honey. In this context, the development of accurate and robust analytical methodologies is of paramount importance for appropriate risk assessment.</p><p><strong>Objective: </strong>This study aimed to develop and optimize a multi-class method for the determination of veterinary drug residues from seven classes (β-lactams, lincosamides, macrolides, quinolones, sulfonamides, tetracyclines, and diaminopyrimidines) using ultrahigh-performance liquid chromatography coupled to time-of-flight mass spectrometry (UHPLC-ToF-MS).</p><p><strong>Methods: </strong>A preliminary assessment was conducted to evaluate the feasibility of solid-liquid extraction, given the complex matrix of compounds present in honey, by analyzing different solvent solutions in acetonitrile, methanol, water, and/or McIlvaine buffer. The subsequent phase of the study involved the optimization of a purification/cleanup method. This was achieved by comparing the efficacy of several solid-phase extraction (SPE) and QuEChERS protocols.</p><p><strong>Results: </strong>The results indicated that the most efficacious extraction were acetonitrile-based solutions, specifically acetic acid 1% and formic acid 0.1% in ACN-H2O (80:20, v/v). Finally, the method with the most optimal analytical performance in terms of recovery and matrix effect entailed an initial extraction step with formic acid 0.1% in ACN-H2O (80:20, v/v), followed by a modified QuEChERS protocol.</p><p><strong>Conclusions: </strong>Acidified organic solvents in combination with QuEChERS method revealed to be the most effective for the determination of antibiotics in honey, ensuring optimum extraction with minimum matrix interference.</p><p><strong>Highlights: </strong>The developed analytical protocol with further detection by high-resolution mass spectrometry will be important to assess health risks and to add value to the consumption of honey and honey-based products.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"339-352"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144059092","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":"Evaluating Physicochemical and Antioxidant Properties of Farm-Fresh and Branded Honey From Bangladesh Using UHPLC-DAD and Multivariate Analysis.","authors":"Jahangir Alam, Manobendra Nath Mohanta, Tamanna Tasnim, Md Azmain Faique, Saidul Islam, Md Athar Ishtiyaq, Mahmudul Bhuiyan, Md Yeasin Prodhan, Md Najim Uddin, Khalid Ali Khan, Md Atikul Islam","doi":"10.1093/jaoacint/qsaf109","DOIUrl":"10.1093/jaoacint/qsaf109","url":null,"abstract":"<p><strong>Background: </strong>The market demand of honey in Bangladesh is rising day by day, but there are concerns about the quality of both the farm-fresh and the branded honey that are available in the market.</p><p><strong>Objectives: </strong>This study evaluates the physicochemical and antioxidant properties of farm-fresh and branded honey from the Bangladeshi market.</p><p><strong>Methods: </strong>The moisture content (MC) and total soluble solid (°Bx) were determined by refractometer; the electrical conductivity (EC) was determined by digital conductivity meter; total phenol (TP), total flavonoid (TF), and HMF were measured by Folin-Ciocalteu, aluminum chloride, and ultrahigh-performance liquid chromatography-diode array detector (UHPLC-DAD), respectively; and antioxidant capacity was assessed by 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method.</p><p><strong>Results: </strong>The results revealed that farm-fresh honey samples exhibited greater content of moisture (>20%), (329.63 ± 1.63 to 986.05 ± 4.81 mg GAE/kg), total flavonoid (95.09 ± 4.99 to 454.73 ± 2.45 mg QE/kg), and antioxidant capacity (IC50 value of DPPH, 47.91 ± 0.19 to 73.66 ± 0.30 mg/mL) compared to branded honey samples (P < 0.05). The HMF amount varied from 0.82 ± 0.00 to 241.24 ± 2.41 (mg/kg), with 42% of branded honey samples exceeding both the Bangladesh Standards and Testing Institution (BSTI) and Codex Alimentarius standard limit (80 mg/kg), whereas all farm-fresh honey samples were within the acceptable limit. In multivariate analysis, the principal component (PC1 & PC2) analysis explains 81.30% of the total variance.</p><p><strong>Conclusions: </strong>Based on compliance with BSTI and Codex HMF limit, farm-fresh honey samples met the acceptable HMF threshold, whereas 42% of branded honey exceeded the standard limit.</p><p><strong>Highlights: </strong>In total, 45 honey samples (farm-fresh, national, international) were analyzed for physicochemical parameters. Farm-fresh honey exhibited higher moisture, phenolics, flavonoids, and antioxidant activity; 42% of branded honeys exceeded HMF standard limits (BSTI and Codex). All farm-fresh samples were within the permissible limit.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"366-376"},"PeriodicalIF":1.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145784173","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}