United States

Recently, the cannabidiol (CBD) products are becoming increasingly available in Japanese food and health markets. In 2024, the Ministry of Health, Labour and Welfare in Japan revised the Cannabis Control Act. This law sets the residual limits of tetrahydrocannabinol (THC) levels in CBD products. Specifically, the residual limits based on Δ9-THC andΔ9- tetrahydrocannabinolic acid (THCA) concentrations were 10 ppm for oil and powder, 1 ppm for various solid samples and 0.1 ppm for liquid samples. In addition, the ministry presents the proposal method for the determination of Δ9-THC and Δ9-THCA based on LC-MS/MS and/or LC-Q/TOF/MS assays. Thus, we investigated that LC-MS/MS assay would be useful, accurate, sensitive and screening evaluation of residual limits of THC levels in common products. Our LC-MS/MS assay of Δ9-THC and Δ9-THCA was developed with reversed-phase short column (running time: 5 min) and ESI-positive/negative mode. Recovery tests with absolute/internal calibrations were good accuracy  and precision of &lt;10%, respectively. In conclusion, our good results were obtained for almost all products, however the recovery rates of water-soluble product were not enough to evaluate these levels using absolute calibrations. On the other hand, we suggest that accurate methods with internal calibration would be useful for the evaluation of residual limits of THC levels in various products.

2025 AOAC INTERNATIONAL Annual Meeting &amp; Exposition

LC-MS/MS Procedure for the Evaluation of CBD Products from Japanese Markets Based on Official Guidelines

Recently, the cannabidiol (CBD) products are becoming increasingly available in Japanese food and health markets. In 2024, the Ministry of Health, Labour and Welfare in Japan revised the Cannabis Control Act. This law sets the residual limits of tetrahydrocannabinol (THC) levels in CBD products. Specifically, the residual limits based on Δ9-THC andΔ9- tetrahydrocannabinolic acid (THCA) concentrations were 10 ppm for oil and powder, 1 ppm for various solid samples and 0.1 ppm for liquid samples. In addition, the ministry presents the proposal method for the determination of Δ9-THC and Δ9-THCA based on LC-MS/MS and/or LC-Q/TOF/MS assays. Thus, we investigated that LC-MS/MS assay would be useful, accurate, sensitive and screening evaluation of residual limits of THC levels in common products. Our LC-MS/MS assay of Δ9-THC and Δ9-THCA was developed with reversed-phase short column (running time: 5 min) and ESI-positive/negative mode. Recovery tests with absolute/internal calibrations were good accuracy  and precision of <10%, respectively. In conclusion, our good results were obtained for almost all products, however the recovery rates of water-soluble product were not enough to evaluate these levels using absolute calibrations. On the other hand, we suggest that accurate methods with internal calibration would be useful for the evaluation of residual limits of THC levels in various products.

#### The AOAC INTERNATIONAL Annual Meeting: A Unique Analytical Science Opportunity

The AOAC Annual Meeting & Exposition provides unparalleled professional development, networking, and collaboration in methods-based science. 

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This multi-faceted “idea incubator” focuses on regulatory changes and emerging food safety issues. A plenary and two breakout sessions feature thought-provoking ideas to identify and meet analytical needs.

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2025 AOAC INTERNATIONAL Annual Meeting & Exposition

The AOAC Annual Meeting & Exposition provides unparalleled professional development, networking, and collaboration in methods-based science.

Molecular confirmation methods have revolutionized microbiological testing, offering speed, sensitivity, and specificity in pathogen and spoilage organism detection. Techniques such as PCR and whole genome sequencing enable rapid identification, bypassing traditional culture-based approaches. However, while these methods enhance detection capabilities, they also introduce challenges in colony confirmation workflows.
One key advantage of molecular confirmation is its ability to detect viable but non-culturable (VBNC) organisms, providing a more accurate assessment of contamination risks. It reduces time to results, allowing for quicker decision-making in food safety interventions. Additionally, molecular methods can differentiate closely related species or serotypes that may be indistinguishable through biochemical or phenotypic tests.
Conversely, molecular approaches present limitations. PCR and sequencing may detect DNA from dead cells, leading to potential false positives. The lack of direct phenotypic confirmation can also create discrepancies between molecular and culture-based results, challenging data interpretation. Moreover, reliance on molecular techniques necessitates specialized training, higher costs, and stringent contamination controls to prevent cross-reactivity or false amplification.
For users, balancing molecular confirmation with traditional colony confirmation is essential. Integrating both approaches provides a comprehensive assessment, ensuring that molecular positives are verifiable through culturing when required for regulatory or investigative purposes. As industry and regulatory bodies evolve, understanding the strengths and weaknesses of each method is critical for developing robust, actionable microbiological testing strategies. This presentation will explore practical considerations, method selection criteria, and real-world applications to optimize confirmation workflows in food microbiology.

The Pros and Cons of Molecular Confirmation and its Impact on Colony Confirmation: A User's Perspective

Bovine lactoferrin (bLF) is commonly fortified in infant formula, pediatric and adult nutritional products for its beneficial impacts on immune system development and gut health. The AOAC Method 2010.10 offers a reliable and accurate solution for determining bLF in powdered infant formula and pediatric/adult nutritional formula, using heparin affinity column extraction and reversed-phase high performance liquid chromatography (RP-HPLC) with ultraviolet detection. We evaluated the advantages of the MaxPeak High Performance Surfaces (HPS) for this analytical method by comparing the results from a conventional column with those from the (same) column incorporated with the MaxPeakTM HPS (XBridgeTM Premier Protein BEHTM C4 Column). We observed more symmetric peak shapes and consistent results, when using the XBridge Premier Protein BEH C4 Column. Additionally, a potential co-elution issue was identified and resolved by implementing a modified gradient elution program. 
MaxPeak, XBridge, and BEH are trademarks of Waters Technologies Corporation.

Benefits of an Inert Surface Technology in the Analysis of Bovine Lactoferrin in Infant Formula and Pediatric/Adult Nutritional Formula

Low-Pressure GC (LPGC technique) has been successfully used for the analysis of pesticide residues, monochloropropane diols (MCPDs), phthalates, arylamines, nitrosamines and alkylamines, as examples. In addition to the applications a variety of different stationary phases have been evaluated to include “5-type”, high phenyl-type, trifluoropropyl and cyano-phenyl columns. Applications have demonstrated using both 0.53mm internal diameter columns as well as 0.32mm internal diameter columns can be successfully used for many different applications. The use of Concurrent Solvent Recondensation Large Volume splitless injection (CSR-LVSI) has been used with LPGC injecting up to 25ul with significant increases in sensitivity and minimal peak distortion. The LPGC techniques provided reductions in run times (up to 3.3x faster runs) and helium consumption reduction (up to 81% less helium used), while keeping an acceptable resolution. This presentation will review applications, use of different column dimensions and phases, in addition will review the ability to inject large volumes to increase sensitivity.

Comprehensive Review of Low-Pressure Gas Chromatography Applications.

Accurate prediction of shelf-life is critical for reducing food waste and boosting consumer confidence in the food industry. Conventional methods like gas chromatography-mass spectrometry and microbial analyses are commonly used to monitor changes in food quality and safety and predict shelf-life. These methods, while accurate, are expensive, time-consuming, and impractical for real-time applications. Hence, a more affordable, simple, and real-time analytical method was developed in this study and tested with samples of soymilk, leveraging the volatile compounds (VOCs) released during spoilage. A novel gold-coated surface-enhanced Raman spectroscopy (SERS)-active fiber was inserted into the headspace of soymilk in capped glass vials. The SERS fiber was incubated in the headspace of the samples at 25ºC for 30 minutes. SERS spectra were acquired using a Raman microscope. Samples were monitored for two weeks. The SERS-active fiber could capture VOCs from soymilk, especially dimethyl sulfide, a well-known bacterial spoilage VOC, during spoilage. Using convolutional neural networks (CNN) models, the SERS spectral changes strongly correlated with changes in pH (R=0.89, RMSE=0.30), microbial growth (R=0.91, RMSE=0.69log10CFU/ml), electrical conductivity (R=0.92, RMSE=0.07mS/cm), zeta-potential (R=0.94, RMSE=1.26mV) and remaining shelf-life (R=0.95, RMSE=1.30days) in the training samples. Further, using SERS spectra from the headspace of a separate set of soymilk samples, the CNN models could predict the remaining shelf-life (R=0.80, RMSE=2.54days) and microbial growth (R=0.73, RMSE=1.14log10CFU/ml). Therefore, the SERS fiber combined with CNN can predict soymilk quality and shelf-life in real-time. Further studies are needed to advance the practical application of this method to more food products.

Real-time prediction of shelf-life of soymilk using a surface-enhanced Raman spectroscopy (SERS) fiber and convolutional neural networks

Thistle is a perennial plant belonging to Asteraceae, which has thorns in its leaves and blooms in summer which grows common in Korea. Thistle has been used for traditional medicine of blood coagulation and hepatoprotection because of known to it contains various biologically active substances along centuries. According to recent research, It has reported to have outstanding antioxidant activity and liver function protection effect. Interests have been increased in thistle. In this study, we extract species of thistles (C. japonicum, C. nipponicum) using extraction solvents((Methanol(MeOH), Dichloromethane(MC), n-Butanol(BuOH), Ethyl acetate(EA), Water), then identify antioxidant activity, total polyphenol content, total flavonoid content. DPPH radical scavenging activity of 1 mg/mL of extract is measured the highest activity(89%) of C. nipponicum and ABTS radical scavenging activity of 0.5 mg/mL of extract is measured the highest activity(94%) of C. nipponicum. The total polyphenol content is the highest in 320mg GAE/g from BuOH fraction of C. nipponicum. Also the total flavonoid content is the highest in 600mg QE/g from BuOH fraction of C. nipponicum. In addition, through 13 Polyphenols and Flavonoids HPLC analysis, EA and BuOH fraction have a high polyphenol contents most of which is chlorogenic acid(EA 40 mg/g, BuOH 65 mg/g), and apigenin, sylingic acid are also contained in extract. 

Comparison of Physiological Activity and Nutritional Components Substance in Extract of Thistle Species

Microdosing is the practice of regularly administering very low doses of a psychoactive substance. Recent studies have indicated microdosing shows promise for the management and treatment of anxiety, depression and other mental health disorders. Emerging research has been focusing on microdosing psilocin and psilocybin, both of which are frequently found in psychedelic mushrooms. With recent legalization of psilocybin and psilocin in certain areas of the United States, there is a growing interest among research and production labs to understand more about the stability and testing of these compounds. This study examines four distinct strains of Psilocybe mushrooms, focusing on the processes of homogenization, extraction efficiency, and extract stability. Primary analysis was performed using LC-UV. A total of seven psychoactive alkaloids commonly found in psychedelic mushrooms were monitored using a simple gradient on a Force Inert Biphenyl column with a total cycle time of 8 minutes. 

Homogenization, Extraction, Stability and Testing by LC-UV of Psychoactive Alkaloids Found in Psychedelic Mushrooms

Seafood substitution poses a significant threat to the global seafood market, driven by increasing consumer demand and fraudulent practices like species mislabeling. Among seafoods, crustaceans are valuable commodities and are particularly vulnerable to substitutionary activities due to their morphological similarities. This study focuses on developing rapid and accurate identification methods to combat seafood fraud. To address this, we developed species-specific C-LAMP and mPCR-LFD assays targeting the COI gene of three mussel species (Perna canaliculus, Mytilus galloprovincialis, and Perna viridis) and three crab species (Portunus pelagicus, Portunus sanguinolentus, and Charybdis feriata). The C-LAMP and mPCR-LFD assay conditions were optimized by varying the temperature and time. The C-LAMP assay demonstrated high accuracy at 63°C for 30 mins. The C-LAMP exhibited high sensitivity for P. viridis and P. canaliculus, with limits of detection of 0.003 and 0.01 ng/reaction, respectively. Similarly, the mPCR-LFD assay achieved high sensitivity, detecting DNA concentrations as low as 0.1 ng, 0.006 ng, and 0.01 ng for blue swimming crab, three spotted crab, and crucifix crab, respectively. Further, in-house validation using various processing methods (boiling, steaming, frying, and canning) using the C-LAMP and mPCR-LFD assay demonstrated high sensitivity and robustness. Out-house validation of crab species revealed a 28% mislabeling rate. Therefore, the C-LAMP and mPCR-LFD assays developed in this study are simple, rapid, and cost-effective for authenticating seafood products. Hence, this novel technology offers a promising solution for routine fishery examinations to stakeholders, industry people, and regulatory authorities, which contributes to seafood safety and authenticity. 

C-LAMP and mPCR – LFD assay: A novel approach for combatting seafood fraud in crustaceans

AOAC 2017.16, for the measurement of total dietary fiber, in alignment with CODEX definitions, achieved final action status in 2020, and replaced AOAC 2009.01 as a CODEX Type 1 method in 2021.  AOAC 2022.01, for the measurement of soluble, insoluble and total dietary fiber is following a similar track with AOAC final action status obtained in 2024 and it is expected to replace AOAC 2011.25 as a CODEX Type 1 method this year. As these methods are implemented across the industry, new insights into analytical challenges continue to emerge. This poster highlights chromatography-related insights , particularly the performance of recommended columns (Waters SugarPak, AOAC 2009.01; Tosoh TSKgel, AOAC 2001.03/2017.16/2022.01) alongside alternative columns sourced globally. A comprehensive carbohydrate dataset was analysed across these columns to evaluate their performance. The comparative chromatographic performance results will be outlined herein.
Additionally, regulatory developments in pet food labelling are driving a transition from crude fiber to dietary fiber analysis, as signalled by the Association of American Feed Control Officials (AAFCO). By 2029, pet food labelling will require dietary fiber determination by approved analytical methods, necessitating a major shift in analytical approaches. To support this transition, we generated relevant data using these methods, assessing their applicability to pet food matrices, the results of which will also be described.

Chromatographic Considerations and Emerging Applications in Dietary Fiber Analysis

The intensive use of pesticides in agriculture has led to the need for rigorous and extensive use of analytical technologies to ensure that there is no impact on human populations.  Maximum residue limits (MRL) are set for regulated residues that define the highest level of a pesticide or mycotoxin residue that is legally tolerated in food such that it is safe for consumers. Often these MRLs are set at low ppb level, to ensure highest safety, very sensitive, robust instrumentation and fast workflows are require quantify these compounds down to their MRL. 
When performing a method with such a vast number of compounds it is important to ensure that the quality of the data is not compromised and that every compound can be effectively quantified. Methods show very high sensitivity to meet different regulations and guidelines. For an assay with over 1500 MRM transitions to analyze, time scheduling of MRMs and instrument switching speed are keys to maintain high quantitative quality. Fast polarity switching time of the instrument ensures good data sampling rates are obtained across the LC peaks for different retention times. Accurate, robust fast quantitation results are presented align with SANTE guideline. Multiple MRMs per analyte also enabled ion ratio monitoring to ensure confident detection. Long batch of several days sample measurements with the representative matrices was run to demonstrate robustness of the method. Improved polarity switching speed allowed for improved quantitation quality for early eluting, polar pesticides. 
 

Ultra-fast MRM acquisition and quantitation of food contaminants in multiple food matrices

Solid Fat Content (SFC) analysis is a cornerstone in the characterization and quality control of fats, fat blends, and finished food products. Understanding the melting behavior of fats is essential for optimizing texture, stability, and processing performance, as well as for timely quality control decision-making. Time-Domain Nuclear Magnetic Resonance (TD-NMR) has emerged as the internationally recognized standard for SFC determination (ISO / IUPAC / AOCS), offering rapid, non-destructive, and solvent-free analysis.
Different SFC workflows based on TD-NMR benchtop technology will be highlighted — including direct and indirect modes for non-invasive determination of solid fat fractions, and parallel and serial tempering techniques for workflow optimization. SFC versus temperature curves obtained from samples of pure and oil-mixed cocoa butter, as well as a fat composite sample are presented. Fully automated SFC workflows, from sample tempering until readings through real time reporting, are highlighted to ensure the strictest compliance with all established international standards, as well as to maximize throughput and results reliability.
Finally, the Dynamic Fat Crystallization (DFC) method, will be discussed as a fundamental technique for the development of new fat substitutes, allowing real time tracking of crystallization behaviors, providing a detailed timeline of initial, main, and final crystallization phases.  Both SFC and DFC techniques empower food manufacturers to ensure consistent product quality, optimize processing conditions, and accelerate R&D cycles. A comparative between DFC curves obtained from samples of cocoa butter and a cocoa butter substitute (palm mid-fraction) are shown.

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The Pros and Cons of Molecular Confirmation and its Impact on Colony Confirmation: A User's Perspective

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