Journal of AOAC International最新文献

{"title":"Development of a Simple and Rapid HPLC-UV Method for Simultaneous Determination of Ciprofloxacin Hydrochloride and Oxytetracycline Hydrochloride in Aqueous Matrix.","authors":"Wan Chi Soon, Ahmad Zuhairi Bin Abdullah, Azam Taufik Mohd Din","doi":"10.1093/jaoacint/qsag096","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag096","url":null,"abstract":"<p><strong>Background: </strong>Reliable and effective methods capable of simultaneously determining ciprofloxacin hydrochloride (CIP) and oxytetracycline hydrochloride (OTC) can be useful when concentrations fall within the validated working range.</p><p><strong>Objective: </strong>This study aimed to develop and validate a simple and rapid reversed-phase high-performance liquid chromatography with ultraviolet detection (RP-HPLC-UV) method for simultaneous determination of CIP and OTC in aqueous matrices.</p><p><strong>Methods: </strong>The separation was performed using an Agilent InfinityLab Poroshell 120 EC-C18 column (150 × 4.6 mm, 4 μm) with a mobile phase consisting of 0.1% formic acid and acetonitrile at a flow rate of 1.0 mL/min. The method was validated for linearity, precision, selectivity, recovery, sensitivity, robustness and system suitability. Matrix performance was assessed using fortified lake-water samples.</p><p><strong>Results: </strong>Linearity over the concentration range of 10-25 mg/L, with R² values of 0.9947 for OTC and 0.9923 for CIP was achieved. The LOD values were 1.40 mg/L for OTC and 1.69 mg/L for CIP, while the corresponding LOQ values were 4.24 and 5.11 mg/L, respectively. Peak-area RSDs were below 1% in both intraday and interday precision studies. Recoveries in fortified lake water ranged from 91.12-109.13% for OTC and 82.88-107.72% for CIP and from 95.84-104.91% and 89.56-112.88%, respectively, in interday analysis. Robustness testing showed that satisfactory chromatographic separation was maintained under the investigated variations in flow rate, column temperature, and detection wavelength.</p><p><strong>Conclusion: </strong>The HPLC-UV approach was applicable for the simultaneous determination of CIP and OTC in aqueous samples containing relatively elevated concentrations of the target antibiotics. The method is not intended for direct trace-level determination in typical environmental waters without an appropriate preconcentration step.</p><p><strong>Highlights: </strong>A simple RP-HPLC-UV method was developed for the simultaneous determination of CIP and OTC. The method is applicable to aqueous samples containing relatively elevated antibiotic concentrations.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149231373","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":"Non-Targeted LC-Q-Orbitrap HRMS Screening of Veterinary Drugs, Pesticides, and Emerging Contaminants in Animal-Derived Foods and Honey.","authors":"Omar Khaled, Lamia Ryad, Fawzy Eissa, Vinoth Kumar Ponnusamy, Mohamed Medhat","doi":"10.1093/jaoacint/qsag095","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag095","url":null,"abstract":"<p><strong>Background: </strong>Veterinary drug residues, pesticides, and chemical contaminants in animal-derived foods pose public health risks, including antimicrobial resistance development and consumer exposure to banned or withdrawn substances. Conventional targeted methods are inherently limited to predefined compounds and may overlook emerging or unexpected contaminants. Many developing countries still lack comprehensive multi-residue surveillance across diverse food matrices, highlighting the need for analytical frameworks adaptable to regulatory laboratories.</p><p><strong>Objective: </strong>To evaluate an LC-Q-Orbitrap HRMS framework combining reference-standard-assisted confirmation with non-targeted screening for the detection and identification of veterinary drug residues, pesticides, and chemical contaminants across multiple animal-derived food matrices.</p><p><strong>Methods: </strong>A streamlined extraction protocol coupled with liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry was applied to 450 retail samples collected in Egypt, covering nine matrices (bovine liver, kidney, meat, heart, raw bovine milk, chicken breast, chicken eggs, honey, and Nile tilapia (Oreochromis niloticus)). Confirmation used 36 certified reference standards; non-targeted screening used Compound Discoverer™ with spectral database matching and in silico fragmentation prediction.</p><p><strong>Results: </strong>Nine compounds were confirmed at Level 1 (confirmed structure via reference standard match) according to the Schymanski identification confidence framework, accounting for 74 detection events, most frequently oxytetracycline (18 detections) and trimethoprim (17 detections). Non-targeted screening revealed 45 additional compounds identified at Level 2a (probable structure based on library spectrum match), including antimicrobials, β-agonists, pesticides, and pharmaceuticals. Phenacetin and tapentadol were each detected in 48 samples, and fenobucarb in 30% of milk samples. Liver and honey showed the greatest diversity of detected compounds.</p><p><strong>Conclusion: </strong>HRMS-based non-targeted screening extends analytical coverage beyond predefined reference-standard panels in multi-matrix food surveillance.</p><p><strong>Highlights: </strong>LC-Q-Orbitrap HRMS detected 54 compounds across nine animal-derived food matrices-nine Level 1 confirmations and 45 Level 2a probable structures-extending chemical surveillance beyond reference-standard panels.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149198766","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":"Current Applications of Hyperspectral Imaging Techniques Combined with Chemometrics for Food Analysis.","authors":"Lili Guo, Huiwen Yu, Teng Wang, Mourad Kharbach","doi":"10.1093/jaoacint/qsag094","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag094","url":null,"abstract":"<p><p>With the growing demand for high-quality and safe food products, innovations in food detection methods are becoming increasingly important. Hyperspectral imaging (HSI) technique combined with chemometrics, has emerged as an efficient, non-destructive, and high-precision approach, offering innovative solutions for food quality and safety analysis in the food sector. HSI technology enables the simultaneous acquisition of spectral and spatial information from food samples. Chemometrics facilitates the extraction of valuable insights from large HSI datasets, allowing for rapid and accurate prediction and classification of food quality parameters. The integration of these two technologies (HSI-chemometrics) has evolved into a promising analytical tool with potential applications across the entire food industry chain. However, its widespread adoption is still constrained by challenges related to model robustness, standardization, and real-world implementation. This paper summarizes recent research progress on the application of HSI-chemometrics in food analysis, focusing on areas such as quantitative analysis of constituents, quality assessment, detection of defects and contaminants, and authenticity verification. Additionally, we summarize current methods for model construction and validation, discuss issues related to model generalization capabilities, and analyze the primary challenges hindering the technology's wider adoption. The goal is to provide a balanced perspective and and technical support for the broader adoption of HSI in food analysis, ultimately contributing to food safety assurance and sustainable development goals.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149198806","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 Three Automated Pretreatment Workstations for Free Silica Determination.","authors":"Jiping Su, Wei Yang, Ruifeng Zheng, Ruirui Dai","doi":"10.1093/jaoacint/qsag093","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag093","url":null,"abstract":"<p><strong>Background: </strong>In China, pneumoconioses accounted for 64.23% of newly reported occupational disease cases in 2024. Silicosis, one of the most common and severe forms of pneumoconiosis, is caused by inhalation of free crystalline silica (SiO₂), highlighting the continued importance of accurate free silica determination. The pyrophosphoric acid (PPA) method (GBZ/T 192.4-2007) predominates in China, but manual operation exposes analysts to hazardous reagents and operator-dependent variability. No comparative evaluation of automated workstations has been published.</p><p><strong>Objective: </strong>To evaluate three automated free silica pretreatment workstations and their agreement with the manual method.</p><p><strong>Methods: </strong>Three workstations (A, B and C) were evaluated with a common protocol in terms of repeatability and trueness on reference and quality control materials, inter-day precision over five days, and agreement with the manual method on field samples by Bland-Altman analysis.</p><p><strong>Results: </strong>Repeatability RSDs (n = 10-12) were 3.5-10.1% at 2.3-5.8% or 9.1%, 2.9-4.8% at 13.0-20.4%, 2.2-2.5% at 58.6-71.6% and 0.5-0.8% at ≥ 99.76% (quartz sand). HorRat exceeded 1.3 in 8 of 13 repeatability or inter-day combinations below 50% free silica and in none at ≥ 60%. Inter-day RSDs were 1.4-4.2% at 15-67% and 0.2-0.8% at 99% free silica. Means lay within the certified value or reference ranges but 0.4-0.9 points below the quartz sand's nominal content. On field samples, mean differences from the manual method were -1.5 to + 0.5 percentage points (not significant), and all 95% limits of agreement lay within ±8 percentage points.</p><p><strong>Conclusion: </strong>All three workstations are safe, reliable alternatives to manual PPA digestion and agreed with the manual method on field samples.</p><p><strong>Highlights: </strong>To our knowledge, the first evaluation of three automated free silica pretreatment workstations with a common protocol; repeatability and trueness characterized over 2-100% free silica; no significant bias relative to the manual method on field samples.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149170596","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":"Research progress on identification and quality control analysis of Eupolyphaga sinensis Walker.","authors":"Jin Han, Lewen Xiong, Zhao Hongwei, Longfei Zhang","doi":"10.1093/jaoacint/qsag091","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag091","url":null,"abstract":"<p><p>Eupolyphaga sinensis Walker is an important animal-derived medicinal material, yet the analytical evidence supporting its authentication and quality control has not been critically mapped across method classes. This scoping review, reported with reference to the PRISMA Extension for Scoping Reviews (PRISMA-ScR), maps methods used for Eupolyphaga sinensis Walker, related or confounding materials, processed products, and exuviae, and distinguishes exploratory proof-of-concept studies from approaches with stronger validation. PubMed, Web of Science, CNKI, Wanfang, and VIP were searched for relevant publications from 1979 to 2025; 81 studies were included and synthesized. Macroscopic and micromorphological examinations can support rapid screening of intact materials but is operator-and specimen-dependent. Powder microscopy can reveal characteristic fragments but has limited discriminatory power. DNA barcoding, species-specific PCR, and SNP-based assays provide stronger evidence of species identity, while mitochondrial PCR-RFLP has been explored for sex determination. Chromatographic and mass spectrometric fingerprinting may support discrimination by sex, geographical origin, processing status, or product type, however, most reported assays lack external or interlaboratory validation. No eligible study established a validated authentication method specifically for Eupolyphaga sinensis Walker exuviae. Overall, no single method meets all identification and quality-control needs. Therefore, it is recommended to adopt a hierarchical workflow that integrates rapid morphological screening, targeted molecular confirmation, and physical and chemical characteristic analysis. On this basis, three priorities should be defined: multi-center verification of gender identification tests, rapid detection of processed products and adulterants, and development of orthogonal verification methods for exuviae.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149139976","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":"Research Progress on Proteomics-Based Detection Techniques for Meat Adulteration.","authors":"Tianshu Li, Huadong Sheng, Yuanxu Shi, Xiangjie Pan, Lulu Wang, Yunxuan Li, Yanxia Zhu, Hongbo Chen, Shuifeng Zhang","doi":"10.1093/jaoacint/qsag057","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag057","url":null,"abstract":"<p><strong>Background: </strong>Meat adulteration is a covert food fraud practice poses considerable safety and ethical risks. This practice challengs traditional detection methods due to biomarker degradation in processed products.</p><p><strong>Objective: </strong>The objective of this review is to summarize recent advancements in proteomics-based techniques and provide a comprehensive methodological and theoretical reference for optimizing meat authenticity verification systems.</p><p><strong>Methods: </strong>The present review compares proteomics with traditional methods and outlines standardized workflows, covering sample preparation, mass spectrometry data acquisition modes, and bioinformatic analysis.</p><p><strong>Results: </strong>Key findings highlight the identification of heat-stable species-specific signature peptides, in addition to the application of these markers across a variety of scenarios, including species identification, tissue-of-origin analysis, and geographical traceability.</p><p><strong>Conclusions: </strong>Proteomics provides high-resolution, multi-target analytical capabilities for complex matrices. Future directions for this field include the translation of technological into biosensors and the global standardization of peptide libraries. Potential leadership for these efforts could come from international organizations, with the aim of advancing industrial and regulatory applications.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149120710","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":"Rapid Detection of Component Contents in Polyphenol-Rich Plant Leaves Based on Near-Infrared Spectroscopy.","authors":"Chen-Peng Wang, Bao-Shan Tang, Yi-Wen Liu, Hong Zhang, Juan Xu, Lan-Xiang Liu, Jin-Ju Ma, Chun-Hua Wu","doi":"10.1093/jaoacint/qsag090","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag090","url":null,"abstract":"<p><strong>Background: </strong>Leaves of Rhus punjabensis var. sinica, Rhus potaninii, Caesalpinia spinosa (Tara), and Phyllanthus emblica (Indian gooseberry), and other species are rich in tannic and gallic acids, which are polyphenols widely utilized in the pharmaceutical, food, and chemical industries. Conventional quantification involves tedious solvent extraction and chemical assays, which are time-consuming, costly, and incapable of simultaneous multi-component analysis.</p><p><strong>Objective: </strong>To enable rapid, non-destructive detection, Fourier transform near-infrared spectroscopy was used to establish quantitative models for synchronous determination of tannic acid, gallic acid, and moisture in polyphenol-rich plant leaves.</p><p><strong>Methods: </strong>Eighty-five leaf samples from six species were collected; reference values were obtained by chemical methods. Near-infrared spectra of plant leaves were fitted to traditional chemical values. Synergy interval partial least squares (SIPLS) selected characteristic intervals to build a partial least squares (PLS) model; performance was assessed by correlation and error metrics.</p><p><strong>Results: </strong>Optimal pretreatments were FD+MSC (moisture), MSC (tannic acid), and FD (gallic acid) with number of principal factors 4, 9, and 6. Characteristic bands: moisture 9403.7-6098.1 and 5450.1-4597.7 cm⁻¹; tannic acid 9403.7-7498.2 and 6102-4246.7 cm⁻¹; gallic acid 9403.7-8451 and 6102-5446.3 cm⁻¹. For moisture, tannic acid, and gallic acid, Rc² = 0.9574, 0.9595, 0.9115; RMSECV = 0.433%, 1.47%, 0.362%; RPD = 4.81, 4.96, 3.36; Rp² = 0.9669, 0.9792, 0.9443; RMSEP = 0.274%, 1.25%, 0.241%. Bias was not significant (P > 0.05), and average RSD < 2%.</p><p><strong>Conclusion: </strong>The results demonstrate that near-infrared spectroscopy combined with chemometric methods demonstrates high predictive capability and practical application value for the quantitative determination of tannic acid, gallic acid, and moisture in polyphenol-rich plant leaves.</p><p><strong>Highlights: </strong>A rapid, non-destructive NIR spectroscopy method was developed for simultaneous quantification of tannic acid, gallic acid, and moisture in polyphenol-rich plant leaves. Optimized chemometric modeling (SIPLS-PLSR) achieved high prediction accuracy (Rp² > 0.94, RPD > 3.0), demonstrating strong robustness and suitability for practical applications.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149083414","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 cadmium, lead and mercury in cannabis flowers by microwave plasma atomic emission spectrometry.","authors":"Mónica Pereira, Ignacio Machado, Florencia Tissot","doi":"10.1093/jaoacint/qsag088","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag088","url":null,"abstract":"<p><strong>Background: </strong>The rapid expansion of legal medicinal and recreational cannabis markets has increased the need for quality control strategies and regulatory monitoring of contaminants in cannabis products. Toxic elements are of particular concern because cannabis plants can accumulate trace metals from environmental and cultivation sources.</p><p><strong>Objective: </strong>To evaluate microwave plasma atomic emission spectrometry (MP AES) as a cost-effective alternative to conventional argon plasma-based techniques for the determination of cadmium (Cd), lead (Pb), and mercury (Hg) in cannabis flowers.</p><p><strong>Methods: </strong>Samples were prepared by microwave-assisted acid digestion using nitric acid. Cd and Pb were determined sequentially by MP AES, while Hg was quantified using a dedicated cold vapor generation coupled to MP AES (CV-MP AES). The analytical methodologies were optimized and validated according to Eurachem guidelines.</p><p><strong>Results: </strong>Limits of quantification were 0.40 µg/g for Cd, 0.18 µg/g for Pb, and 0.015 µg/g for Hg. Accuracy was evaluated using reference materials and spiked samples, with recoveries of 95-101% and 81-90%, respectively, and relative standard deviations below 15%. Good linearity was achieved (R² > 0.999). The methodologies were applied to 30 commercial cannabis flower samples, where Cd and Hg were not detected, and Pb was quantified at 2.12 ± 0.13 µg/g.</p><p><strong>Conclusions: </strong>MP AES and CV-MP AES provide adequate accuracy and precision for the determination of Cd, Pb, and Hg in cannabis flowers. The proposed methodologies represent a practical, cost-effective, and sustainable alternative to argon-based plasma techniques for routine quality control and compliance testing of cannabis products.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149043069","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 Risk-Based Framework to Optimize Microbiological Testing of Primary Packaging Materials.","authors":"Prada-Ramírez Harold Alexis, Cely-Veloza Willy, Ericsson Coy-Barrera, Humberto Zardo, Guerrero-Bonilla Jeny, Castro-Prieto Libardo Elías, Montes-Tamara Juan Pablo, Gloria-Inés Lafaurie, Raquel Gómez-Pliego","doi":"10.1093/jaoacint/qsag089","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag089","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Risk-based approaches are increasingly encouraged to optimize microbiological control strategies for non-sterile pharmaceutical materials. Primary packaging materials represent a critical interface, yet routine microbiological testing is often applied uniformly without considering material characteristics or historical performance.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Aims: &lt;/strong&gt;This study aimed to develop and evaluate a risk-based framework to support reducing routine microbiological testing for non-sterile primary packaging materials by integrating water activity (Aw), historical microbiological data, and intrinsic material and process attributes.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Common primary packaging materials, including PVC, PVC-PVDC, aluminum foil, PET, HDPE, LDPE, and polymeric tubes, were evaluated. Water activity was measured and compared against an upper specification limit (Aw ≤ 0.60). Historical microbiological performance was assessed using at least 20 consecutive compliant batches without out-of-specification (OOS) or out-of-trend (OOT) results. A one-sided process capability analysis (Cpk) was performed to characterize Aw variability relative to the specification limit.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;Historical data supported sustained control across materials, with at least 20 consecutive compliant batches and Aw values consistently ≤ 0.60. Most materials showed low variability and operated with a substantial margin relative to the Aw limit, whereas HDPE and LDPE exhibited comparatively higher variability. These findings were not associated with evidence of microbiological proliferation within the evaluated dataset. Cpk values quantitatively described observed performance and were interpreted as supportive indicators given the limited dataset.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;These findings support a risk-based strategy to optimize the frequency of microbiological testing. Historical microbiological performance represents the primary basis for reduced testing, while Aw and material attributes provide complementary evidence. This approach enables optimization of testing under controlled conditions without replacing microbiological control, in alignment with ICH Q9(R1), ICH Q6A, and relevant compendial guidance.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Highlights: &lt;/strong&gt;A risk-based framework that integrates water activity, historical microbiological data, and material attributes is proposed to optimize microbiological testing of non-sterile primary packaging materials. All evaluated materials showed Aw ≤ 0.60, consistent with conditions unfavorable to microbial proliferation, while historical data (≥20 compliant batches) provided the primary evidence of sustained control. Differences in Aw variability were observed, particularly for HDPE and LDPE, highlighting the importance of material-specific characteristics in risk assessment. Overall, the approach supports the reduction-rather than replacement-of microbiological testing under control","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149043071","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":"Edible Fish as a Source of Klebsiella pneumoniae: Incidence, Virulence, Biofilm Formation, Antimicrobial Resistance and Control Strategies.","authors":"Ahamed Basha Kusunur, Prasanna Kumar Medidi, Sradha Dinesh Kumar, Alekhya Nakka, Himabindu Chigilipalli, Rudrappa Gattu, Visnuvinayagam Sivam, Anandan Rangasamy, Murugudas Vaiyapuri, Madhusudana Rao Badireddy","doi":"10.1093/jaoacint/qsag087","DOIUrl":"https://doi.org/10.1093/jaoacint/qsag087","url":null,"abstract":"<p><strong>Background: </strong>Fish sold in retail markets are prone to contamination with pathogenic bacteria. Investigations on occurrence of Klebsiella pneumoniae, its characterization in seafood are essential for the protection of consumer health.</p><p><strong>Objective: </strong>The present study investigated the incidence of K. pneumoniae in fish and characterized their virulence profile, biofilm-forming ability, antimicrobial resistance and their control using water-soluble chitosan and postbiotics.</p><p><strong>Methods: </strong>In total, 144 fish and processing water samples from fish retail markets of Visakhapatnam, India were screened for presence of K. pneumoniae using MacConkey agar and amplifying khe gene. Virulence characterization (entB, mrkD, kfu, allS, rmpA, ybtS, iutA) antibiotic susceptibility test (disc diffusion assay against 28 antibiotics) biofilm-forming ability and anti-K. pneumoniae activity of water-soluble chitosan (microtiter plate assay) were studied.</p><p><strong>Results: </strong>Eleven fish samples showed presence of K. pneumoniae. All the isolates possessed entB gene, however, 91% of the isolates carried mrkD and kfu genes. Antibiogram studies showed that all isolates were multidrug resistant (MDR), and their Multiple Antibiotic Resistance (MAR) index ranged from 0.19 to 0.39. Resistance genes detected in K. pneumoniae were bla  SHV (91%), bla  TEM (27%), sul1(18%), tetA (9%), tetD (9%), and catA2 (9%). Majority of K. pneumoniae isolates were moderate biofilm formers (63.6%) and the biofilm-forming ability was potentiated by supplementation with lactose (100%) followed by fructose (90.9%) and ribose (90.9%) sugars. Water-soluble chitosan (1%) showed effective control of all K. pneumoniae.</p><p><strong>Conclusion: </strong>The present study showed majority of the K. pneumoniae isolates from seafood were found to be virulent, multidrug-resistant, ESBL producers, and moderate biofilm formers indicating potential health risk to fish consumers. Water-soluble chitosan showed a significant inhibitory effect on growth of all K. pneumoniae isolates.</p><p><strong>Highlights: </strong>The detection of virulent and MDR K. pneumoniae in edible fish meat, albeit at a low level, poses a significant threat to consumers.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149019232","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}