Journal of AOAC International最新文献

{"title":"New Insights into Probiotic Enumeration in a Complex Food Matrix: Flow Cytometry and Plate Count Correlation.","authors":"Gonzalo Saiz-Gonzalo, Orlaith O'Connell, Shima Joy, Seamus McSweeney, Nooshin Vahedi Kia, André Brodkorb, Sinéad B Bleiel","doi":"10.1093/jaoacint/qsag009","DOIUrl":"10.1093/jaoacint/qsag009","url":null,"abstract":"<p><strong>Background: </strong>Accurate probiotic enumeration ensures the dose delivered to the mammalian host. Plate count (PC) detects only culturable cells and can underestimate the total number of viable cells, particularly when some cells are injured or dormant and do not form colonies.</p><p><strong>Objective: </strong>To qualify a flow cytometry assay reporting active fluorescent units (AFU) and to compare counts with the PC method for micro-encapsulated Lacticaseibacillus rhamnosus GG in a snack product.</p><p><strong>Methods: </strong>Following AOAC INTERNATIONAL, International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH), and United States Pharmacopeia (USP) principles, precision, accuracy, ruggedness, specificity, and robustness of plate count and flow cytometry methods were evaluated for the first time in a commercial yogurt-bite snack product.</p><p><strong>Results: </strong>Both methods fulfilled the pre-specified performance targets (RSD ≤15%; AFU recovery 100-104%; live/dead R2 ≥0.95). Analyst-to-analyst differences were not significant. Flow cytometry spike recoveries were 100-104% across targets. Live/dead mixtures tracked linearly (R2 ≥0.95). Across matched samples, AFU and CFU values were equivalent within a pre-specified ±0.5 log band; batch means typically differed by ≤0.2 log with no systematic bias.</p><p><strong>Conclusion: </strong>The qualified flow cytometry method provides same-day counts and enumerates all membrane-intact cells, complementing plate counts while meeting analytical performance criteria. Findings endorse the use of AFU in tandem to CFU assessment for probiotic quantification, label accuracy and estimation of microbiome-relevant dose in snack and finished-product matrixes.</p><p><strong>Highlights: </strong>Flow cytometry provided same-day results and enumerated all membrane-intact cells for label verification in snack matrixes.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"556-565"},"PeriodicalIF":1.7,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146097770","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":"The SureFast® Listeria 3plex ONE Kit and the SureFast® PREP Bacteria Kit in a Variety of Foods and Environmental Samples: Multi-Laboratory Validation, First Action 2025.04.","authors":"Kateland M Lanzit, M Joseph Benzinger, Benjamin Bastin, Erin Crowley, Patricia Meinhardt, Sara Altmeyer, Svetlana Woitkowiak, Caroline Knoll, Martin Mehl","doi":"10.1093/jaoacint/qsag015","DOIUrl":"10.1093/jaoacint/qsag015","url":null,"abstract":"<p><strong>Background: </strong>The SureFast® Listeria 3plex ONE Kit uses the real-time polymerase chain reaction (PCR) technique for multiplex detection of Listeria species and Listeria monocytogenes in foods and environmental samples. The SureFast Listeria 3plex ONE Kit can also be used with the SureFast PREP Bacteria Kit to provide a second DNA extraction option and can be analyzed on multiple thermocyclers.</p><p><strong>Objective: </strong>The kit was evaluated using 25 g test portions in an unpaired study design for a single matrix, ready-to-eat smoked deli turkey, as part of a harmonized validation protocol with MicroVal Certification.</p><p><strong>Methods: </strong>The matrix was compared to the International Organization for Standardization Technical (ISO) 11290-1:21017 reference standard; 17 participants from 16 laboratories located within Austria, Germany, Italy, Spain, and the United States were solicited for the multi-laboratory validation (MLV), with only 10 participants submitting data. Three levels of contamination were evaluated: an uninoculated control level (0 colony-forming units (CFU)/test portion), a low inoculum level (0.2-2 CFU/test portion), and a high inoculum level (2-10 CFU/test portion). Statistical analysis was conducted according to the Probability of Detection (POD) statistical model.</p><p><strong>Results: </strong>Results obtained for the low inoculum level test portions produced a dLPOD value with 95% confidence interval of -0.01 (-0.04, 0.02). The dLPOD results indicate equivalence between the candidate method and reference method for the matrix evaluated and the method demonstrated acceptable multi-laboratory reproducibility as determined in the MLV. False positive and false negative rates were determined for the matrix and produced values of 0.0% and 0.8%, respectively.</p><p><strong>Conclusions: </strong>Based on the data generated, the method demonstrated equivalence to the ISO reference method in a MLV study.</p><p><strong>Highlights: </strong>The SureFast ONE 3plex method is a rapid and reliable alternative for the multiplex detection of Listeria species and Listeria monocytogenes in a broad range of foods and environmental surfaces.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"576-588"},"PeriodicalIF":1.7,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13337140/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147617231","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":"Preparation of Functionalized Immunomagnetic Beads and Their Application in the Detection of AFB1 in Corn, Oat, and Peanut.","authors":"Huihui Hao, Mengke Wu, Xiaolei Zhao, Jianjun Li, Jinxing He","doi":"10.1093/jaoacint/qsag002","DOIUrl":"10.1093/jaoacint/qsag002","url":null,"abstract":"<p><strong>Background: </strong>Aflatoxin B1 (AFB1) is widespread in various kinds of food and poses a serious threat to health when it enters the human body via the food chain.</p><p><strong>Objective: </strong>This study aimed to establish a rapid and sensitive method for the detection of AFB1 and then combine it with HPLC for the quantitative analysis of AFB1 content.</p><p><strong>Methods: </strong>In this work, hydrothermal and sol-gel methods were employed to prepare aminated magnetic nanoparticles featuring strong magnetism and excellent dispersion. Magnetic nanoparticles bound with antibodies can specifically capture AFB1 and then be combined with HPLC for the quantitative analysis of AFB1 content. Moreover, the key factors influencing the extraction efficiency, such as buffer type, adsorption time, elution time and volume, were optimized.</p><p><strong>Results: </strong>The samples were spiked with AFB1 at low, medium, and high concentration levels of 5.0, 10.0, and 20.0 μg/kg, respectively. The established method exhibits good linearity within the range of 0.1-100 µg/kg, and the detection limit is as low as 0.09 µg/kg (S/N = 3). The recoveries in real samples ranged from 79.33 to 111.51%, with all RSDs being less than 11.16% (n = 3 for each level).</p><p><strong>Conclusion: </strong>In conclusion, a rapid and efficient analytical method for detecting AFB1 in corn, peanut, and oat based on functionalized immunomagnetic beads (IMBs) coupled with high-performance liquid chromatography with a fluorescence detector (HPLC-FLD) have been successfully developed. The method exhibited excellent sensitivity, featuring a low method detection limit of 0.09 μg/kg, high accuracy, and remarkable precision. In comparison to traditional sample preparation techniques, this IMB-based approach presents substantial advantages in terms of simplicity, rapidity (completing sample processing within approximately 30 min), and cost-effectiveness.</p><p><strong>Highlights: </strong>The IMBs possess excellent magnetic separation capabilities, which effectively streamlines the pre-processing steps.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"496-504"},"PeriodicalIF":1.7,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145947165","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":"Food Adulteration by Granular Formulation of Carbamates for Animal Poisoning: Chromatographic Determination and Vibrational Microspectroscopy.","authors":"Fernanda Ziegler Reginato, Karol Andriely de Vargas Paier, Maria Odete da Silva Dalan, Gustavo Andrade Ugalde, Sailer Santos Dos Santos, Marcelo Barcellos da Rosa, André Valle de Bairros","doi":"10.1093/jaoacint/qsaf092","DOIUrl":"10.1093/jaoacint/qsaf092","url":null,"abstract":"<p><strong>Background: </strong>Carbamates have been used in intentional poisoning cases to cause death in both humans and animals. Granulated material containing carbamate pesticides is often applied to the victim's food, water, or drink. Toxicological analysis is essential to confirm such cases.</p><p><strong>Objective: </strong>This study aims to identify and quantify carbamates (methomyl, aldicarb, propoxur, carbofuran, and carbaryl) present in granular formulations used to adulterate food, through the application of chromatographic and vibrational microspectroscopic techniques.</p><p><strong>Methods: </strong>Thin-layer chromatography (TLC), liquid chromatography with diode array detector (LC-DAD), gas chromatography-mass spectrometry (GC-MS), attenuated total reflectance Fourier transform infrared (ATR-FTIR), and Raman microspectroscopy were performed. An aliquot of granular formulation was manually separated for chromatographic methods; dilutions were employed as test material treatment, while vibrational microspectroscopy techniques used raw material for toxicological analysis. A material similar to a shredded sausage containing suspected granular formulation was evaluated in chromatographic tests, and carbofuran was determined.</p><p><strong>Results: </strong>The proposed TLC methodology for identifying carbamates proved unfeasible due to insufficient chromatographic separation. The LC-DAD method was fully validated with success for methomyl, aldicarb, propoxur, carbofuran, and carbaryl, revealing a concentration of 744 µg/g of carbofuran in the suspected material, which was further confirmed by GC-MS. Raman microspectroscopy was not precise enough for carbamate identification, but ATR-FTIR demonstrated high specificity and robustness in identifying carbamate pesticides present in the test material.</p><p><strong>Conclusions: </strong>This study allowed determining the profile of carbamate pesticides in granular material when present in adulterated food/feed. To isolate the granules manually reduces the carrying of matrix interferents from food and/or feed. Furthermore, the techniques applied require only the raw material or its diluted versions, avoiding steps, high consumption of organic solvents, and reducing analysis time and cost.</p><p><strong>Highlights: </strong>LC-DAD was successfully validated for multiple carbamates. Minimal test material prep reduces solvent use, analysis time, and costs.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"439-448"},"PeriodicalIF":1.7,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240600","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":"Monitoring Emerging Contaminants in Soil and Household Dust Samples by Non-Targeted Analysis in the Greater Miami Area, Florida.","authors":"Luciana Teresa Dias Cappelini, Olutobi Daniel Ogunbiyi, Vinícius Guimarães Ferreira, Carolina Cuchimaque Lugo, Monica Beatriz Perez, Mymuna Monem, Florence George, Piero Gardinali, Daniel M Bagner, Natalia Quinete","doi":"10.1093/jaoacint/qsaf012","DOIUrl":"10.1093/jaoacint/qsaf012","url":null,"abstract":"<p><strong>Background: </strong>Emerging organic contaminants are increasingly present in the environment, with several pollution sources. Soil and household dust are critical pathways for human exposure, posing significant health risks, especially for vulnerable children.</p><p><strong>Objective: </strong>The objective of this study was to identify and characterize the presence of emerging contaminants in soil and house dust samples in Miami-Dade and Broward counties, Florida, evaluating how seasonal, socioeconomic, and geographic factors influence the distribution of these pollutants.</p><p><strong>Methods: </strong>Soil and house dust samples were collected from 49 families twice a year between May 2022 and February 2024. Samples were processed by accelerated solvent extraction (ASE) and analyzed by liquid chromatography-high-resolution mass spectrometry (LC-HRMS). Chemical identification was performed using Compound Discoverer (CD) software.</p><p><strong>Results: </strong>The predominant compounds in soil were personal care products (PCP)/surfactants, industrial products, and pesticides in the dry season, while pharmaceuticals were found in higher abundance in the rainy season. The most abundant compounds in household dust were pharmaceuticals, industrial products, and PCP/surfactants in the dry season, while pesticides stood out in the rainy season. Geographically, the central Miami region presented a wider diversity of contaminants in dust and more PCP/surfactants in soil, while the northern region presented greater contamination by pharmaceuticals and industrial products in soil and more surfactants in dust. Overall, lower-income regions showed greater diversity and abundance of pollutants in dust and soil.</p><p><strong>Conclusions: </strong>These results suggest that the distribution of organic pollutants in soil and house dust is influenced by seasonal, socioeconomic, and geographic factors. These differences highlight the need for adaptive environmental policies and integrated strategies to mitigate the risks associated with exposure to these pollutants in urban environments, protecting public health.</p><p><strong>Highlights: </strong>Seasonal variations significantly influence the distribution of emerging contaminants in soil and household dust. Socioeconomic and geographic factors contribute to distinct contamination patterns across Miami-Dade and Broward.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"449-467"},"PeriodicalIF":1.7,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143485075","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":"Rapidly and Simultaneously Determining the Total Polysaccharides Content, Total Flavonoids Content, and Antioxidant Activity in Polygonatum cyrtonema Hua by NIR Spectroscopy and Chemometrics.","authors":"Xuanxuan Fan, Xian Hu, Weiting Wang, Naifu Chen, Naidong Chen, Jingwen Hao, Le Yan, Jun Xiao, Yu Zhong, Manman Zhang, Zhengjun Xie","doi":"10.1093/jaoacint/qsag003","DOIUrl":"10.1093/jaoacint/qsag003","url":null,"abstract":"<p><strong>Background: </strong>Polygonatum cyrtonema Hua (PCH) is widely used in traditional Chinese medicine. The components and activity are important for simultaneous quality evaluation in PCH.</p><p><strong>Objective: </strong>This study aimed to establish an accurate, rapid, and comprehensive quality evaluation method by near-infrared spectroscopy (NIR) for forecasting the total polysaccharides content (TPsC), total flavonoids content (TFC), and antioxidant activity (AA) in PCH.</p><p><strong>Methods: </strong>PCH samples were used to establish a model of TPsC, TFC, and AA in PCH by NIR combined with partial least-squares regression (PLS). To enhance the accuracy of the models and remove non-essential variables, we used multiple spectral preprocessing methods and multiple chemometrics to analyze the processed full spectrum, such as competitive adaptive reweighted sampling partial least-squares regression (CARS-PLS), moving-window partial least-squares regression (MW-PLS), and interval random frog partial least squares regression (iRF-PLS). The remaining samples were used to complete external validation.</p><p><strong>Results: </strong>Satisfactory prediction results of PLS models combined with chemometrics were obtained. The optimal chemometric of TPsC, TFC, and AA selected 157, 105, and 134 variables, respectively. For the TPsC, TFC, and AA models, which incorporated optimal spectral preprocessing and chemometrics, the root-mean-square error of calibration (RMSEC, %) values were 0.743, 0.069, and 0.136, while the R2 values were above 0.95. The root-mean-square error of prediction (RMSEP, %) for these models were 1.00, 0.074, and 0.153, while R2 values were above 0.90. The remaining 20 samples were used to complete external validation, which confirmed the preeminent comparability and ability of the proposed method.</p><p><strong>Conclusion: </strong>NIR combined with chemometrics provides an effective, fast, and nondestructive approach for evaluating quality via multiple indicators of PCH and provides a meaningful reference for evaluation of herbal medicine quality.</p><p><strong>Highlights: </strong>NIR combined with chemometrics offers an effective, quick, nondestructive way to evaluate herbal medicine quality with multiple indicators.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"596-606"},"PeriodicalIF":1.7,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145949192","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":"Optimization of Extraction Buffer Conditions for Improved Heparin Affinity-Based Quantification of Bovine Lactoferrin in Various Dairy Products.","authors":"Tomoaki Fukushima, Keisuke Yamamoto, Yukino Sato, Tatsuya Nojima, Mirei Odaka, Norichika Nishida, Hiroshi Ochi","doi":"10.1093/jaoacint/qsag001","DOIUrl":"10.1093/jaoacint/qsag001","url":null,"abstract":"<p><strong>Background: </strong>Bovine lactoferrin (bLF) has been supplemented to a variety of foods, particularly dairy products, to promote human health. Reliable analytical methods are required for accurate quantification of bLF in foods. Isolation of bLF from foods using a heparin column and quantification via HPLC have been reported (heparin method); however, its applicability remains limited.</p><p><strong>Objective: </strong>To develop a broadly applicable method for determining bLF in dairy products by optimizing the extraction buffer conditions used in the heparin method.</p><p><strong>Method: </strong>bLF was spiked into different dairy products and extracted using buffers with or without additives, such as urea, sodium chloride, EDTA, and Tween 20. These additives reduced interactions between bLF and dairy components and improved the interaction of bLF to the heparin column, with the buffer pH adjusted to 6.0. The extracted bLF was isolated using a heparin column and quantified via C4 reversed-phase HPLC using an external standard calibration curve.</p><p><strong>Results: </strong>The optimized extraction buffer improved bLF recovery rates across all tested dairy product matrixes.</p><p><strong>Conclusion: </strong>An improved extraction buffer was developed that enhanced the applicability of bLF quantification using the heparin method. This optimized extraction buffer enabled reliable recovery of bLF from diverse and complex dairy product matrixes.</p><p><strong>Highlight: </strong>The optimized extraction buffer significantly improves the recovery and quantification of bLF in various dairy products, providing a versatile solution for accurate bLF analysis.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"589-595"},"PeriodicalIF":1.7,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13337142/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145936855","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":"Single-Laboratory Validation of Simultaneous Determination of Aflatoxins in Nutraceuticals following immune-affinity Column Cleanup and Liquid Chromatography Tandem Mass Spectrometry Analysis.","authors":"Saravanan J, Santhosh Kumar J Urumarudappa, Vijay Bommuluri, Ashutosh Kumar Mittal, Peter Chang, Gary Swanson","doi":"10.1093/jaoacint/qsag059","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag059","url":null,"abstract":"<p><strong>Background: </strong>Aflatoxins B1, B2, G1, and G2 are highly carcinogenic mycotoxins frequently detected in nutraceutical products. Due to their toxicological significance and the complex composition of nutraceutical matrices, sensitive and validated analytical methods are required to ensure regulatory compliance, product safety, and routine surveillance.</p><p><strong>Objective: </strong>This study aimed to develop and perform single-laboratory validation of a reliable and sensitive method for the simultaneous determination of aflatoxins B1, B2, G1, and G2 in nutraceutical products, in accordance with AOAC Appendix F.</p><p><strong>Method: </strong>Samples were extracted using an organic solvent system and purified through aflatoxin-specific immunoaffinity columns to reduce matrix interference. Quantitative analysis was performed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Method validation evaluated linearity, limits of detection (LOD) and quantitation (LOQ), precision (repeatability and intermediate precision), and accuracy across three representative nutraceutical matrices.</p><p><strong>Results: </strong>The method showed linearity for all analytes, with coefficients of determination (R2) between 0.9984 and 0.9989. The LOD and LOQ values ranged from 0.1268 to 0.6170 µg/kg and from 0.2733 to 1.1290 µg/kg, respectively. Precision studies demonstrated acceptable repeatability and intermediate precision, with relative standard deviations ≤20%. Accuracy assessment across beverage and protein-based nutraceutical products yielded recoveries between 88% and 119% at multiple fortification levels, meeting AOAC performance requirements for mycotoxin analysis.</p><p><strong>Conclusions: </strong>The validated LC-MS/MS method is sensitive, accurate, and precise for the determination of aflatoxins B1, B2, G1, and G2 in complex nutraceutical products. The method is fit for purpose of routine quality control, regulatory testing, and surveillance of aflatoxins in nutraceutical matrices.</p><p><strong>Highlights: </strong>Optimized sample extraction and immunoaffinity cleanup enabled sensitive LC-MS/MS determination of aflatoxins B1, B2, G1, and G2. The method was validated for linearity, LOD, LOQ, sensitivity, precision, and accuracy, and was found to be suitable for routine aflatoxin estimation in nutraceutical products.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148341674","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 Bromoform in Seaweed, Oil, and Animal Feed by GC-MS/MS: AOAC Official Method 2026.01, First Action.","authors":"Saeedeh Taheri, Siong H Tan, Ashwini Jadhav, Robert B S Burgess, Greg Aylward, Sara Labaf","doi":"10.1093/jaoacint/qsag061","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag061","url":null,"abstract":"<p><strong>Background: </strong>Bromoform is the bioactive compound present in Asparagopsis seaweed responsible for methane reduction in ruminants. Although several analytical methods are available, no AOAC Official Method exists.</p><p><strong>Objective: </strong>A study was performed to validate a GC-MS/MS-based method for determination of bromoform in fresh and freeze-dried seaweed, canola oil containing bromoform (oil herein), and liquid and dry cattle feed.</p><p><strong>Methods: </strong>The method was evaluated according to AOAC guidelines for calibration curve validity, selectivity, extraction protocol robustness, and matrix and extract stability. Repeatability, recovery, and limits of detection and quantification were also determined for each matrix.</p><p><strong>Results: </strong>The calibration curve was linear over the range 0.000124-0.020 µg/mL (R2= 0.999), and residual analysis demonstrated suitability of the linear model. There was no interference from compounds similar to bromoform (dibromomethane, dichloromethane, dibromochloromethane, and dibromoacetic acid). The analyte was identified by comparison of retention time and ion abundance profiles between calibration standards and unknown samples. The LOQs were 0.057 mg/g, 0.423 mg/g, 0.033 mg/g, 0.022 mg/g, and 0.041 mg/g for fresh seaweed, freeze-dried seaweed, oil, liquid feed, and dry feed, respectively. Bromoform recovery among matrixes across multiple fortified concentration levels was 93-104%, and repeatability was 0.75-5.4%. The method was robust to minor changes in extraction conditions. This method was demonstrated to be accurate and precise with or without the inclusion of an internal standard.</p><p><strong>Conclusion: </strong>The GC-MS/MS method for bromoform determination presented in this report was shown to be fit for purpose and has been accepted as a First Action AOAC Official Method  SM.</p><p><strong>Highlights: </strong>This method is the first bromoform analysis method to successfully complete the AOAC validation process and will serve to standardize the determination of bromoform in feed ingredients and finished supplements. The adoption of this method is timely given the recent interest in methane reduction in cattle using feed fortified with bromoform.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334966","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":"ICRF-Assisted Box-Behnken Design and Optimization for Rapid UPLC-PDA Determination of Dorzolamide Hydrochloride and Timolol Maleate in an Ophthalmic Preparation.","authors":"Erdal Dinç, Eda Büker","doi":"10.1093/jaoacint/qsag060","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag060","url":null,"abstract":"<p><strong>Background: </strong>Simultaneous optimization of chromatographic resolution and analysis time constitutes a significant analytical challenge in multicomponent pharmaceutical analysis, as resolution-driven optimization strategies may improve peak separation without providing explicit control over retention time, often resulting in unnecessarily prolonged runtimes. To address this limitation, an optimization strategy based on a Box-Behnken experimental design was implemented in conjunction with the Improved Chromatographic Response Function (ICRF), which integrates separation quality and analysis time within a single mathematical objective function.</p><p><strong>Objective: </strong>This strategy describes the development of a rapid, chemometrically optimized UPLC-PDA method for the simultaneous determination of dorzolamide hydrochloride (DH) and timolol maleate (TI) in a commercial ophthalmic preparation.</p><p><strong>Method: </strong>A Box-Behnken experimental design and optimization approach was employed in combination with an Improved Chromatographic Response Function (ICRF), which integrates resolution, peak overlap, peak width, and runtime into a single composite objective function. This strategy enabled short runtime (or short retention time of analytes in a chromatogram) while preserving adequate peak separation.</p><p><strong>Results: </strong>Under the optimized conditions, complete chromatographic separation was achieved within 3 min using a BEH C18 column and a mobile phase consisting of acetonitrile and 4 × 10-4 M CCl3COOH (60:40, v/v) at a flow rate of 0.32 mL/min with detection at 275 nm. The method demonstrated excellent linearity over the range of 5.0-40.0 µg/mL (r > 0.999), with limits of detection of 0.51 µg/mL for DH and 0.61 µg/mL for TI. Mean recoveries were 99.9% and 99.5% for DH and TI, respectively, with satisfactory precision and robustness.</p><p><strong>Conclusion: </strong>The proposed ICRF-assisted optimization approach provided high-resolution separation within minimal runtime and was successfully applied to the routine quality-control analysis of a commercial ophthalmic formulation.</p><p><strong>Highlights: </strong>The study demonstrates the effectiveness of composite response-based chemometric optimization in enhancing analytical efficiency in pharmaceutical drug analysis.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148321093","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}