United States

The National Institute of Standards and Technology (NIST) Dietary Supplement Laboratory Quality Assurance Program (DSQAP) was established in 2007 as part of a collaboration with the National Institutes of Health (NIH) Office of Dietary Supplements (ODS). The DSQAP was created to facilitate improved measurement capabilities of the dietary supplement (DS) and natural product communities. Participating laboratories are able to assess their in-house analytical methods through a variety of studies including toxic and nutritional elements, fat- and water-soluble vitamins, fatty acids, organic contaminants, and marker compounds in samples that represent DS ingredients or finished products. These studies are designed to address analytical challenges in the community, e.g. emerging ingredients and products, material content, and lowering limits of detection. Insights into community measurement performance and analytical needs derived from DSQAP studies also informs NIST and NIH ODS in the production and maintenance of references materials for DSs. Results from DSQAP studies can be utilized to identify problematic matrices or analytes within the DS measurement communities. These results can also determine the need for new reference materials and/or which analytes should be targeted to aid in the standardization of measurements. This presentation will highlight results from elemental studies from previous and current DSQAP exercises that will benefit the DS analytical communities through the continued improvement of nutritional and toxic element characterization in DS ingredients and the development and maintenance of reference materials at NIST.

2025 AOAC INTERNATIONAL Annual Meeting &amp; Exposition

NIST Dietary Supplement Laboratory Quality Assurance Program (DSQAP): Overview of Elemental Studies and Development of Reference Materials

#### The AOAC INTERNATIONAL Annual Meeting: A Unique Analytical Science Opportunity

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The 2025 AOAC INTERNATIONAL Annual Meeting & Exposition was held in person in San Diego, USA.

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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.

Whether for ethical, dietary, health, or social reasons, there are increasing numbers of people globally who follow a vegan or vegetarian diet.  The food industry is aware of the rise in popularity of meat-free food and is actively investigating processes and products that can help meet the demand.
To ensure that non-meat based protein or cultured protein products are safe for human consumption, manufacturers must comply with Good Manufacturing Practice (GMP).  The FDA currently implements comprehensive methods for the elemental analysis in food digests.  This presentation will discuss the sample preparation using microwave assisted decomposition and the associated elemental analysis by ICP-MS.  The validation of the method will also be discussed to ensure data accuracy and instrument performance.

Elemental Analysis of Plant-Based Alternative Meat Sample and Cell Culture Solutions by ICP-MS

Heavy metals contaminate our food and drinking water by soil and air pollution via pesticides, industrial waste or accidental release. Plants uptake metals from the environment, depositing them throughout the plant including flowers, pollen and fruit. In some cases, these are beneficial minerals necessary for bone structure and cognitive health, but when grown in contaminated soil heavy metals can contaminate our food sources as well. Some forms of these heavy metals have more toxic forms, like methylmercury or hexavalent chromium when compared to other mercury or chromium forms. ICP-MS alone cannot identify concentrations of these more harmful species, as a total value is collected for the element.
We have paired separation by HPLC followed by ICP-MS to better understand the concentrations of the more harmful forms contaminating our food and water. Demonstrated will be examples including the separation of inorganic mercury and methyl mercury in 5 minutes, followed by detection using ICP-MS. All tested elements were tested for sensitivity at 10% and 20% of the Food Safety and Standards Authority of India and reported excellent repeatability and RSD below 3%.

Improving Risk Assessment and Mediation of Heavy Metals in Foods Using Speciation by HPLC Coupled ICP-MS

The trend towards a healthier diet and lifestyle has expanded the functional mushroom market and continues to grow exponentially. Finished products now come in a variety of forms and flavours. However, the boom of the medicinal mushroom industry has led to an urgent need for the development of fit-for-purpose methods to support authenticity claims and improve consumer trust.   
The key active component found in mushroom products is 1,3/1,6-β-glucan. While the measurement of 1,3:1,4-β-glucan found in cereals and the 1,3/1,6-β-glucan found in yeast can be achieved using a selective enzymatic hydrolysis approach, an equivalent method is not available for the measurement of 1,3/1,6-β-glucan in mushrooms, due to the complex structure presented. Instead, an indirect method involving subtraction of enzymatically determined a-glucan content from an acid hydrolysis/chemo-enzymatic derived “total glucan” measurement, can be employed as described by McCleary et al. in 2016. Although this method readily allows for the measurement of β-glucan in mushroom products which are contaminated with α-glucans comprising starch, maltodextrins and/or sucrose, other components may present challenges.
As such, a number of assay parameters have been thoroughly investigated, and the results of this study are described herein. 

Measurement of β-glucan in Mushrooms and Derived Products: Critical Evaluation and Improved Methodology

Co-author:Arun Krishnamurthy & Amber Thelen, Purity-IQ Inc
Functional mushroom products are available in various forms, including whole vouchers, extracts, blends, and mycelia fermented grains, each adding complexity to product analysis. While targeted methods provide valuable species-level identification, they may not fully capture the broader chemical profile that reflects product quality, which can be influenced by manufacturing practices, processing, and geographical origin. To complement these targeted techniques, robust, non-targeted analytical tools are essential for a comprehensive evaluation of both species-authenticity and product quality — key components of consumer trust.
Nuclear Magnetic Resonance (NMR) spectroscopy offers both targeted and non-targeted chemical profiling capabilities, providing deep insight into the species identity, purity, and quality of functional mushroom products. When combined with multivariate statistical analysis, NMR enhances fraud detection and supports quality control by identifying chemical variations that might otherwise go unnoticed. A validated NMR spectral library, developed using authentic materials obtained from credible sources and cross-verified with other orthogonal methods, enables robust species discrimination, extract characterization, and quality assessment.
NMR’s versatility strengthens existing analytical frameworks by offering enhanced detection of adulteration, consistency verification, and chemical characterization. This study demonstrates the practical application of NMR metabolomics and statistical analysis in fungal product evaluation, showcasing its value as a powerful, routine testing method for authenticity, purity, and overall quality in the mushroom supplement market.

Beyond Species Identification: Leveraging NMR for Comprehensive Quality Assurance in Mushroom Supplements

Fennel fruit consists of the dried ripe fruit of Foeniculum vulgare Mill. (Fam. Apiaceae); it has been used in traditional medicines for a wide range of ailments related to digestive, endocrine, reproductive, and respiratory systems. The marketed fennel fruit is sometimes adulterated with fruits of Carum carvi L., Cuminum cyminum L. and Anethum graveolens L. or is much lower quality due to unsuitable processing and storage.
USP is developing Fennel Fruit quality standards to help protect customers from using incorrect and low-quality fennel fruit. The USP Fennel Fruit monographs provide analytical quality control procedures and acceptance criteria with suitable reference standards.
The monograph contains identification methods utilizing GC, HPTLC and botanical characteristics, and a GC assay method. Both GC and HPTLC identification methods in the monograph can detect anethole, a major component of fennel fruit. The three potential confounders do not contain anethole, which can efficiently differentiate the fennel fruit from the potential confounders. The GC assay can quantify content of anethole, limonene, fenchone, and estragole concurrently. Test results indicated the content ratio of anethole to fenchone in fennel fruit is much different from bitter fennel (Foeniculum vulgare Mill. var. vulgare) fruit a variety of fennel used in Europe. The USP Laboratory tested some commercial fennel products using the monograph methods and found that some of the products did not contain any related components in both GC and HPTLC chromatograms, implying that the fennel fruit maybe an exhausted product which may not be acceptable for use as a dietary ingredient.

USP Monograph Methods Differentiate Fennel Fruit from the Potential Confounders and Closely Related Species

The objective of this work was to develop methods for the analysis of antibiotics and veterinary drugs in milk listed by Food Safety and Standards Authority of India (FSSAI) using a PerkinElmer QSight 420 LC-MS/MS system with a StayClean™ ion source. To improve spiked recoveries and better method performance, two separate methods were developed for the analysis of over 30 analytes in milk due to their different polarities. For polar analytes, a mixed mode LC method was used for analyte separation and a trichloroacetic acid/acetonitrile mixed solution was used for protein precipitation. For other less polar analytes, a reversed phase UHPLC method was used for analyte separation and an acidified acetonitrile extraction was applied for sample preparation. For both methods, EDTA was used to reduce analyte-metal (i.e., calcium) ion complex formation and d-SPE was performed to remove fatty matrix using end-capped C18 sorbent. To reduce matrix effects, matrix-matched calibration with isotopic mass dilution method was performed. Both methods were validated by spiking different concentrations of analytes in a milk sample matrix. The recoveries of most analytes are between 70 to 120%. The results demonstrated that all the analytes could be determined with LOQ below the tolerance limits set by FSSAI. In addition, StayClean ion source allows continuing injections of large amounts “dirty” samples with virtually no instrument cleaning and maintenance.  An instrument robust test result dataset with continuing injections of 25,500 samples will be presented. This methodology can be extended to analysis of similar analytes in milk and milk products regulated by other governmental agencies.

Stay Clean LC-MS/MS for Determination of Antibiotics and Veterinary Drugs in Milk

A collaborative study was conducted to validate the suitability of an LC-MS/MS method for quantifying 128 residual veterinary drugs in six food matrices: honey, milk, egg, beef, pork liver, and lard. Twelve laboratories participated, including seven governmental, four commercial, and one incorporated foundation. Standard compounds, extraction powders, and matrices were provided to minimize material-related variations. Laboratories spiked samples with 5 ppb, 25 ppb, and 50 ppb levels, and reported recoveries along with chromatograms. At the 5 ppb spiking level, a significant number of compounds showed recoveries below 60% or above 125%, with the highest occurrence in milk (82 compounds), followed by honey (70), lard (68), pork liver (59), beef (54), and egg (49). However, at the 25 ppb level, the number of compounds with recoveries outside the range of 70%–120% significantly decreased, with pork liver showing the highest number (8  compounds), followed by honey (4), and only 2 compounds in other matrices. At the 50 ppb level, the numbers outside the range further decreased, with pork liver (5  compounds), beef (4), egg (4), lard (3), milk (1), and honey (1). These results indicate that the LC-MS/MS method performs optimally at a limit of quantification (LOQ) of 25 ppb, as the number of compounds with unacceptable recoveries significantly decreases at this level. By setting the LOQ at 25 ppb and filtering out poorly performing compounds, this method can be effectively optimized to serve as a reliable and robust approach for veterinary drug residue analysis in complex food matrices.

Collaborative Test of an LC-MS/MS Method for the Determination of 128 Veterinary Drugs in Six Food Matrices

Biofilm formation on food contact surfaces poses significant challenges to food safety due to increased resistance to disinfectants and risk of cross-contamination. This study aims to develop a machine learning–enhanced nanosensor array for the classification and prediction of bacterial biofilms on food processing surfaces, particularly stainless steel. A fluorescence-based sensor array composed of 2D nanoparticles and ssDNA probes was designed to capture compositional and developmental features of biofilms formed by four pre-screened species: Escherichia coli, Staphylococcus aureus, Salmonella enterica, and Pseudomonas fluorescens. Biofilms were cultivated in 96-well plates and on stainless steel coupons across three timepoints (days 1, 3, and 5). Sensor responses were recorded and analyzed using three machine learning algorithms—random forest (RFC), support vector classification (SVC), and multilayer perceptron (MLP)—to classify species and biofilm maturity (12 total classes). Data preprocessing included normalization, imputation, and stratified train-test splitting. Model performance was evaluated using accuracy, F1 score, and ROC-AUC with a one-vs-rest strategy. Preliminary results indicate species-specific fluorescence patterns and distinct clustering in PCA analysis. The sensor array is expected to achieve over 90% classification accuracy, with slightly lower performance for early-stage biofilms. This approach enables rapid, label-free biofilm identification and maturity assessment, offering a scalable solution for real-time monitoring of microbial contamination in food environments. The findings support the integration of advanced sensing and machine learning techniques for biofilm control strategies in food safety applications.

Machine Learning–Integrated Nanosensor Array for Predictive Biofilm Classification on Food Processing Surfaces

Introduction 
Food allergens are proteins that can trigger an immune response in allergic individuals such as nausea, vomiting, asthma, or in extreme cases anaphylactic shock.  One of the most common food allergies in children is a milk allergy.  Typically caused by cow’s milk, this allergy can also come from sheep, goats, buffalo, and other mammals.  Symptoms include wheezing, vomiting, hives, digestive problems, and even anaphylaxis.  Fortunately, studies have shown that most children will outgrow a milk allergy.
 
Objective 
To evaluate the performance of two rapid lateral flow milk assays on various market basket products. 
 
Materials and Methods 
10 milk-containing (n=6) and non-milk-containing (n=10) products were analyzed.  Milk containing samples were analyzed neat and diluted to extinction.  Non milk containing samples were spiked using a qualified milk spike stock increasing in 1 ppm increments until a positive result was achieved. In addition, milk spikes were swabbed off a stainless-steel surface (n=3 per level) to verify the detection level of a swab for each test kit.  
 
Results 
Both rapid milk test kits performed exceptionally well.  For non-milk containing samples, spiking data showed recovery down to 5 ppm total food (1.8 ppm protein) on all samples.  With market basket samples containing milk, the test kits detected milk below kit detection levels to ensure kit sensitivity within various matrices, down to 3 ppm total food (1.1 ppm protein).  In addition, the surface swabbing analysis indicated that the test kits can detect low levels of milk on food contact surfaces.   
 
Significance 
To ensure the safety of consumers with a milk allergy, it is of high importance to have a quick and reliable test for food manufacturers to use at their facilities.  The data shows that Neogen has two such rapid test kits allowing food manufacturers to have cost effective options when analyzing for milk allergens. 

An Evaluation of Rapid Lateral Flow Assays for Total Milk Detection

Per- and polyfluoroalkyl substances (PFAS) are known potential contaminants in eggs which are commonly consumed by humans. Recently, the European Union has set regulatory limits for four PFAS compounds (PFOA, PFOS, PFNA, PFHxS) in eggs, and interest in testing food for PFAS is growing in the United States. Individuals who consume eggs from backyard chickens may assume they are safer or healthier yet remain unaware of potential PFAS contamination due to unknown exposures in the chickens feed, bedding, water, and environment – potential routes through which PFAS can enter the eggs they produce.
This study compares PFAS concentrations in cage-free store-bought eggs and eggs from backyard chickens, which may be exposed to a broader range of environmental factors and dietary variations. A workflow for analysis of this challenging matrix will be presented which utilizes an automated sample preparation method that reduces analyst involvement, limits variability, and enhances throughput. The automated sample extraction process takes less than 15 minutes per sample, and the automated solid-phase extraction (SPE) system can process up to 8 samples simultaneously in under 70 minutes. This method uses dual-phase Graphitized Carbon Black and Weak Anion Exchange SPE cartridges to clean up challenging samples to ensure precise and repeatable results across samples. The automation of both the sample preparation and SPE limits variability, reduces method time, and increases throughput. Ultimately, this study aims to provide a clearer understanding of PFAS contamination in both commercially and locally sourced eggs, contributing valuable insights for public health and food safety policies.

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