Journal of AOAC International最新文献

{"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}

{"title":"A Green UPLC-UV Method for L-Ascorbic Acid Determination Based on a Biodegradable Chelating Agent and Synergistic Hydrophobic-Electrostatic Interactions.","authors":"Wen Gu, Zuyan Fang, Bingwen Lin, Liang Zhou, Jianzhen Li, Zhengfei Lu, Peter Chang, Gary Swanson","doi":"10.1093/jaoacint/qsag056","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag056","url":null,"abstract":"<p><strong>Background: </strong>Ascorbic acid (Vitamin C) is the most widely consumed dietary supplement, which necessitates extensive quantitative testing of these products for batch release and post-market surveillance. However, most methods for ascorbic acid determination rely on hazardous reagents. It is therefore essential to adopt a green analytical chemistry (GAC) approach to evaluate and improve these methods, thereby mitigating their adverse environmental impact of analytical activities.</p><p><strong>Objective: </strong>This study aimed to develop a green ultra-performance liquid chromatography (UPLC) method for determining L-ascorbic acid in different types of dietary supplements, using biodegradable reagents and minimizing organic solvent consumption.</p><p><strong>Method: </strong>A phosphate solution (pH 3.0) containing 5 mmol/L oxalic acid (OA) was used for sample preparation. A 2-µL aliquot of the ascorbic acid extract was injected into a UPLC system. Separation was performed on an AX-C18 column (2.1 mm i.d. × 100 mm, 1.9 µm) with a mobile phase of 0.4% (v/v) phosphoric acid in water. Isocratic elution was carried out at a flow rate of 0.2 mL/min for 3 minutes. Ascorbic acid was detected at 245 nm and quantitated against standard solutions of L-ascorbic acid.</p><p><strong>Results: </strong>OA demonstrated effectiveness in stabilizing ascorbic acid comparable to that of conventional chelators, while offering superior environmental sustainability. When coupled with a UPLC system, this method achieved a low limit of quantitation (0.02 μg/mL) and excellent recoveries (98.3%-101.2%). Analytical Eco-Scale and AGREE metrics were used to evaluate the greenness of this method, indicating its excellent environmental performance.</p><p><strong>Conclusions: </strong>This newly developed method demonstrated robust performance across diverse sample matrices, providing an eco-friendly and highly efficient approach for determining vitamin C in dietary supplements and health foods fortified with L-ascorbic acid.</p><p><strong>Highlights: </strong>Minimized organic solvent usage, faster turnaround time, and improved environmental compatibility make this a precise, efficient, and green method for vitamin C determination.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148285059","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":"Quantification of 24 Inorganic Impurities in Vismodegib and Idelalisib Anti-Cancer Drug Substances by Using ICP-MS.","authors":"Raj Kumar Kodishala, Kousrali Sayyad, Sreedhar Gundekari, Naresh Konduru, Venkateswara Rao Bhoomi, Leela Prasad Kowtharapu, Mohan Varkolu, T B Patrudu","doi":"10.1093/jaoacint/qsag055","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag055","url":null,"abstract":"<p><strong>Background: </strong>Metal impurities in pharmaceutical products can originate from various sources, including manufacturing machinery, catalysts used during drug synthesis, and even the container-closure systems.</p><p><strong>Objective: </strong>The objective of the present study is to accurately quantify trace levels of 24 metal impurities in the active pharmaceutical ingredients of the anticancer drugs vismodegib and idelalisib using an inductively coupled plasma mass spectrometer.</p><p><strong>Method: </strong>The diluent used in the analysis consists of a mixture of 3% nitric acid and 1% hydrochloric acid, while argon serves as the carrier gas. Helium is utilized as the collision gas at a flow rate of 4.3 mL/min. The plasma gas flow rate is maintained at 16 L/min, and the spray chamber temperature is controlled at 2.0 °C to ensure optimal stability during analysis. The dwell time for each metal impurity is set to 0.3 s, and the instrument is precisely tuned in helium mode to achieve accurate and interference-free detection of trace metal impurities.</p><p><strong>Results: </strong>No interference was observed from the calibration blanks, confirming the method's selectivity and accuracy. The sample recoveries were satisfactory, falling within the USP acceptance range of 70-150%.</p><p><strong>Conclusion: </strong>The analytical procedure was performed in accordance with USP <233> and ICH Q3D(R2) guidelines, and all results were found to be within the established acceptance criteria, thereby validating the reliability and compliance of the method.</p><p><strong>Highlights: </strong>To date, no research studies have been reported on the determination of 24 elemental metal impurities in the drug substances vismodegib and idelalisib. Therefore, the present research aims to develop a unified and reliable analytical method for the quantification of these metal impurities in the active pharmaceutical ingredients of both drugs, ensuring compliance with regulatory standards and enhancing the overall quality and safety of the pharmaceutical products.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148274399","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":"LC-MS/MS Method for Per- and Polyfluoroalkyl Substances (PFAS) Analysis in Food Products, First Action 2025.10.","authors":"Xanthippe Theurillat, Claudia Mujahid, Bjorn Eriksen, Antoine Levêques, Thierry Delatour, Pascal Mottier, Ashley Griffin, Andrew Savage","doi":"10.1093/jaoacint/qsag054","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag054","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Per- and Polyfluoroalkyl Substances (PFAS) are synthetic compounds widely used due to their water, grease, and dirt repellent properties. They are persistent and resistant to degradation, therefore they can be found in the environment, the food chain, and in human fluids. The European Union (EU) has established limits of quantification (LOQs) for four PFAS in produce, seafood, meat, eggs, fish oil, milk, and ready-to-eat food for infants and young children and set maximum levels (MLs) for the same analytes in eggs, fish, and meat products.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Objective: &lt;/strong&gt;The study describes a quantitative LC-MS/MS method for the determination of 57 PFAS in food. The 55 analytes are quantified by isotopic dilution while two of the compounds, namely Capstone A and Capstone B are quantified using 2-level standard addition. Seven of the compounds have only one transition, which need another technique for their confirmation in case of a positive result in a sample. The scope comprises analytes included in the AOAC INTERNATIONAL Standard Method Performance Requirements (SMPRs®) 2023.003, cited in the EU Recommendation 2022/1431, and analytes surveyed by service laboratories in Europe or mentioned in the literature.Representative samples from several food categories (baby food puree, milk-based infant formula (powder), full cream milk powder, fish meat and fish oil, whole egg, soluble coffee, pet food and edible offal) were validated. Targeted performance requirements for LOQ, recovery and precision were as defined per AOAC SMPR 2023.003.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Method: &lt;/strong&gt;The protocol involves an acetonitrile/water extraction. For 55 PFAS analysis in all matrices, a solid phase extraction (SPE) method is applied. Detection is conducted by LC-MS/MS and isotopic dilution is applied for quantification. Confirmation for single-transition analysis is performed by LC-HRMS. For infant formula and baby food, the extract is diverted to two workflows, one for the 55 PFAS and one for Capstone A & Capstone B, where a diluted aliquot is analyzed by LC-MS/MS and quantified by the standard addition procedure.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The method was single laboratory validated in the nine food matrices with results generally in agreement with the criteria of the AOAC SMPR 2023.003, in terms of LOQs, recovery and precision. LOQs were generally in the low ng/kg range, and recoveries typically fell within 70-120%, with acceptable repeatability and intermediate precision. The performance of the method was further demonstrated by analyzing 27 additional food matrices.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;The method described covers a wide range of food matrices and analytes compares favorably with most of the requirements of the SMPR 2023.003, it contains as well a larger scope of analytes compared to the SMPR 2023.003, giving an advantage of searching other potential sources of PFAS contamination. Following independent scientific review","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148229825","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":"Comprehensive Comparison of ISO-GUM and Monte Carlo Simulation for Uncertainty Assessment in HPLC Statin Quantification with single and multi-point calibrations.","authors":"Hamza Belmir, Hasnaa Haidara, Rajae Aitlhaj, Taoufiq Saffaj, Bouchaib Ihssane","doi":"10.1093/jaoacint/qsag053","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag053","url":null,"abstract":"<p><strong>Background: </strong>Reliable measurement uncertainty estimation is essential for building trust in analytical results and ensuring they meet regulatory expectations. In HPLC chromatography, the chosen approach for uncertainty estimation can strongly influence both the results and their interpretation.</p><p><strong>Objective: </strong>This study compares two widely used strategies, the ISO-GUM method and Monte Carlo simulation applied with either single-point or multi-point calibration, for the simultaneous quantification of four statins at the National Laboratory for Drug Control (LNCM).</p><p><strong>Methods: </strong>In the single-point approach, all sources of uncertainty were combined into one global model, while the multi-point approach evaluated variability across different calibration levels. Both strategies were applied within the ISO-GUM framework and through Monte Carlo simulation, allowing a side-by-side comparison of the resulting uncertainty budgets.</p><p><strong>Results: </strong>The single-point method tended to underrepresent the variability present in actual measurements. In contrast, the multi-point method, especially when paired with Monte Carlo simulation, delivered uncertainty estimates that better reflected real-world analytical conditions. The ISO-GUM framework combined with multi-point calibration also proved to be a robust and practical option for routine use.</p><p><strong>Conclusions: </strong>Multi-point calibration, whether combined with Monte Carlo simulation or the ISO-GUM method, offers a more representative and reliable way to assess uncertainty in HPLC statin analysis.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148229792","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":"Zinc, Selenium, and Iodine Biofortification of Hard-Dough Biscuits Using Agronomically Biofortified Whole Wheat Flour: Mineral Retention, Physicochemical Properties, and Sensory Acceptability.","authors":"Yaşar Karaduman, Erdinç Savaşlı, Oğuz Önder, Arzu Akın, Seda Tetik, Tuğba Ak","doi":"10.1093/jaoacint/qsag051","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag051","url":null,"abstract":"<p><strong>Background: </strong>Micronutrient deficiencies (zinc, selenium, iodine) affect billions globally, necessitating sustainable food-based solutions. Agronomic biofortification of staple crops offers a promising strategy, but effective translation into palatable, processed foods remains critical.</p><p><strong>Objective: </strong>To develop hard-dough biscuits using whole wheat flour from wheat agronomically biofortified with zinc alone (FZn biofortified flour) or zinc, selenium, and iodine (FZnSeI), and to quantify mineral enrichment, technological properties, and sensory acceptability.</p><p><strong>Methods: </strong>Whole wheat flours from biofortified wheat were substituted at 0, 25, 50, 75, and 100% levels into a standard biscuit formulation. Mineral content (Zn, Se, I) was analyzed by ICP-OES. Physicochemical properties (hardness, color, moisture, ash, protein, fat), gluten strength (GlutoPeak), and sensory attributes (5-point scales, n = 12 trained panelists) were evaluated.</p><p><strong>Results: </strong>FZn flour contained 30.6 mg/kg zinc; FZnSeI flour contained 25.2 mg/kg Zn, 152.6 µg/kg Se, and 211.4 µg/kg I. Dose-dependent increases were observed in biscuits. At 100% substitution, FZn biscuits showed 30.57 mg/kg Zn (106.3% increase over control). FZnSeI biscuits contained 25.22 mg/kg Zn, 152.62 µg/kg Se (576.5% increase), and 211.38 µg/kg I. A 100 g serving of triple-fortified biscuit provided 22.9, 27.7, and 14.1% of the adult RDA for Zn, Se, and I, respectively. Sensory evaluation showed that 75-100% substitution levels maintained overall acceptability despite greater textural firmness in FZn biscuits.</p><p><strong>Conclusion: </strong>Whole wheat flour from agronomically biofortified wheat enables effective production of palatable, nutrient-dense hard-dough biscuits with significantly elevated Zn, Se, and I levels, offering a scalable strategy to combat hidden hunger.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148166055","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":"Revisiting the relationship between reproducibility and concentration: a new, data-driven statistical model.","authors":"Stefan Ehling, Paul Wehling, Philip A Haselberger, Lawrence H Pacquette","doi":"10.1093/jaoacint/qsag048","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag048","url":null,"abstract":"<p><strong>Background: </strong>The Horwitz equation has long been used to model the relationship between reproducibility relative standard deviation (RSDR) and analyte concentration. However, modern chromatographic and spectroscopic methods routinely achieve substantially better reproducibility than predicted, particularly for homogeneous nutritional matrices. A new data-driven model is needed to characterize current analytical performance.</p><p><strong>Objective: </strong>To develop and evaluate a new statistical model describing the relationship between reproducibility standard deviation (sR) and analyte concentration (C), using data from 961 analyte-matrix combinations across 62 analytes.</p><p><strong>Methods: </strong>Previously published multi-laboratory study data were reanalyzed by regressing log-transformed reproducibility standard deviation against log-transformed concentration, with both expressed as mass fractions. Ordinary least squares regression was performed following an approach analogous to that used previously for dietary fiber data.</p><p><strong>Results: </strong>A strong linear relationship was observed between log(sR) and log(C) (R2 = 0.98) across concentrations spanning 3.7E-10 to 2.0E-2. Deviations from the model were largely attributable to specific analytical methods or analyte characteristics-particularly AOAC methods 2014.08, 2011.14, and 2015.06. The model indicates that RSDR increases very gradually with decreasing concentration, well below values predicted by the Horwitz equation except at very high concentrations.</p><p><strong>Conclusion: </strong>Reproducibility precision for modern analytical techniques applied to homogeneous nutritional matrices can be effectively modeled by a simple linear relationship between log(sR) and log(C). This empirical model captures current analytical performance trends but is not intended as a method performance criterion.</p><p><strong>Highlights: </strong>The new regression model describes a linear relationship between log(sR) and log(C) across 961 data points, spanning eight orders of magnitude. RSDR increases far more gradually with decreasing concentration than predicted by the Horwitz model. Deviations are primarily linked to specific methods and analytes.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148166075","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":"ATP Bioluminescence for Rapid Environmental Monitoring in Non-Sterile Pharmaceutical Manufacturing: A Quality by Design Approach.","authors":"Prada-Ramírez Harold Alexis, Cely-Veloza Willy, Ericsson Coy-Barrera, Guerrero-Bonilla Jeny, Castro-Prieto Libardo Elías, Colmenares Nicolás, Gloria-Inés Lafaurie, Sonia Orjuela, Montes-Tamara Juan Pablo, Humberto Zardo, Gómez-Pliego Raquel","doi":"10.1093/jaoacint/qsag050","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag050","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Rapid and reliable environmental monitoring is critical in non-sterile pharmaceutical manufacturing to ensure contamination control and product quality. Conventional culture-based methods require extended incubation times and may delay decision-making. ATP bioluminescence has emerged as a rapid alternative; however, its implementation as a quantitative method requires appropriate validation in accordance with regulatory guidelines, particularly considering its limitation as a non-specific indicator of total biological residues rather than exclusively viable microorganisms.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Aims: &lt;/strong&gt;To validate an ATP bioluminescence-based method as a rapid alternative to conventional culture-based techniques for environmental microbiological monitoring in non-sterile pharmaceutical manufacturing areas, in accordance with USP &lt;1223&gt; and a Quality by Design framework.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;The method was evaluated using a pooled inoculum of Salmonella spp., Escherichia coli, Aspergillus brasiliensis, and Candida albicans, applying a commercial ATP bioluminescence system (Clean-Trace™) under controlled environmental surface conditions. Validation parameters included equivalence of results, linearity (R2 ≥ 0.9025), working range (1-500 CFU), precision (RSD), specificity, accuracy (recovery 75-125%), and limits of detection and quantification (&lt;10 CFU). Statistical analyses included regression modeling and Bland-Altman agreement assessment (p &gt; 0.05).&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;Results demonstrated strong quantitative agreement between ATP-derived signals (RLU) and CFU-based methods, supported by correlation coefficients (R2 ≥ 0.9025) and recovery values ranging from 90% to 114%. Bland-Altman analysis confirmed that differences between methods were within acceptable limits of agreement. A QbD-based robustness assessment evaluated the impact of sanitizing agents on the luciferin-luciferase reaction, identifying sodium hypochlorite and benzalkonium chloride as sources of transient signal interference, which stabilized after approximately 5 min. Cleaning procedures effectively removed both residual and microbial ATP from contaminated surfaces, achieving levels below 10 CFU and &lt;15 RLU across multiple stainless steel types.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;The ATP bioluminescence method met predefined USP &lt;1223&gt; validation acceptance criteria, including recovery (75-125%), statistical agreement (p &gt; 0.05), and precision (RSD within acceptable limits), and demonstrated equivalence to conventional microbiological techniques. The method enables rapid and reliable detection of surface contamination, reducing analysis time from several days to seconds and supporting real-time decision-making in manufacturing environments.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Highlights: &lt;/strong&gt;This study presents a validated ATP bioluminescence method that enables real-time environmental monitoring in non-sterile pharmaceutical manufacturing, si","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148166062","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 Fatty Acids at the sn-2 Position of Triacylglycerol Molecules in Infant Formula and Adult/Pediatric Nutritionals by Enzymatic Transesterification, Single-Laboratory Validation Study: First Action 2025.09.","authors":"Daniëlla Holt, Yomi Watanabe, Pierre-Alain Golay, Hans Cruijsen, Francesca Giuffrida, Hugues Griffon","doi":"10.1093/jaoacint/qsag046","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag046","url":null,"abstract":"<p><strong>Background: </strong>The positional distribution of the fatty acids on the triacylglycerol influences uptake of calcium, fat, and energy. The positional distribution of breast milk is similar, but not identical, to that of cow and goat milk, and is different to that of vegetable oil. Often, in infant formula a mixture of milk fat with certain vegetable oils is used, to mimic the positional distribution of breast milk. A method for the analysis of fatty acids at the sn-2 position in infant formula and adult/pediatric nutritionals by enzymatic transesterification is presented.</p><p><strong>Objective: </strong>The aim of this study is to demonstrate the suitability and applicability of the method to determine fatty acids at the sn-2 position of triacylglycerol molecules in infant formula and adult/pediatric nutritionals by enzymatic transesterification.</p><p><strong>Methods: </strong>The sample is extracted to isolate the fat, which is treated with a regiospecific enzyme to remove the sn-1(3) positions of the triacylglycerol, resulting in 2-monoacylglycerols. These are then separated using solid phase extraction, transesterified to fatty acid methyl esters, and analyzed using gas chromatography with flame ionization detection.</p><p><strong>Results: </strong>The relative standard deviation repeatability (RSDr) and intermediate precision (RSDiR) were in the range of 1.2-5.2% (RSDr) and 2.0-13.3% (RSDiR). The accuracy was assessed by spiking experiments and model triglycerides, and recoveries were on average 95% (spiked addition) and 97% (model triglycerides). The limit of quantification (LOQ) was found at 0.003 g/100g on reconstituted product.</p><p><strong>Conclusion: </strong>The SLV demonstrates that it meets the criteria set in the Standard Method Performance Requirements (SMPRSM) 2022.004.</p><p><strong>Highlights: </strong>A method to determine the content of fatty acids at the sn-2 position by use of an enzyme with good 1(3) regiospecificity was developed. The method complies to international and governmental regulations. The method was evaluated by the Expert Review Panel on Nutrient Methods, and granted first action status.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148145871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An Integrated QuEChERS LC-MS/MS Method for Screening, Confirmation, and Quantification of Nitroimidazole Residues in Food.","authors":"Hoi-Yan Fiona Shiu, Kong-Chi Lau, Ho-Pan Michael Yau","doi":"10.1093/jaoacint/qsag047","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag047","url":null,"abstract":"<p><strong>Background: </strong>Nitroimidazoles are prohibited veterinary drugs, for which sensitive and reliable analytical methods are required for regulatory residue control in food, particularly across diverse and complex matrices.</p><p><strong>Objective: </strong>This study aimed to develop an integrated QuEChERS-LC-MS/MS method for screening, confirmatory identification, and quantitative determination of nitroimidazole residues within a single harmonized analytical workflow.</p><p><strong>Methods: </strong>Samples were prepared using a unified QuEChERS procedure followed by dispersive solid-phase clean-up and LC-MS/MS analysis with isotope-labelled internal standards. Screening was conducted using a predefined screening target concentration (STC) as a validation reference level. Samples meeting relative retention time requirements and exhibiting responses exceeding a conservative screening cut-off were classified as suspected positives and subjected to confirmatory evaluation based on ion ratio criteria, followed by quantitative determination within the same analytical run. The method was validated for muscle, offal, fat, milk, egg, and aquatic products. Decision limits (CCα) and detection capabilities (CCβ) were established in accordance with regulatory guidelines.</p><p><strong>Results: </strong>Excellent linearity was obtained (r2 ≥ 0.99) over the range of 0-2.5 µg/L. Mean recoveries ranged from 84 to 116%, with RSDs 0.1-8.1%. Limits of quantification were 0.2-0.4 µg/kg across all matrices, and matrix effects were adequately controlled (91-110%).</p><p><strong>Conclusion: </strong>The validated method provides a robust and harmonized analytical workflow suitable for screening, confirmation, and quantification of nitroimidazole residues across a wide range of food matrices, supporting regulatory residue control.</p><p><strong>Highlights: </strong>A single harmonized QuEChERS-LC-MS/MS workflow was developed for the screening, confirmatory identification, and quantitative determination of four nitroimidazoles and their metabolites in animal and aquatic foods. The method applies an STC-based decision strategy with confirmatory identification criteria and was validated across multiple matrices, achieving low LOQs. Performance was evaluated in accordance with Commission Implementing Regulation (EU) 2021/808 and verified through international proficiency testing.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148145853","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}