Journal of AOAC International最新文献

{"title":"Analysis of Bovine Immunoglobulin G in Milk-Based Commodities: Liquid Chromatography-Tandem Mass Spectrometry vs. Affinity Liquid Chromatography.","authors":"Stefan Ehling, Ujwal S Patil, Paul J Baker","doi":"10.1093/jaoacint/qsaf095","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf095","url":null,"abstract":"<p><strong>Background: </strong>Current analytical methods for bovine immunoglobulin G (IgG) have certain limitations in terms of selectivity, reproducibility, and applicability to complex matrices. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) using peptide-based quantitation and a suitable calibrant can potentially overcome some of these limitations.</p><p><strong>Objective: </strong>Commercial bovine IgG standards and colostrum powder were evaluated for suitability as calibrants. The feasibility of using a single peptide marker for quantitation of total IgG by LC-MS/MS was assessed in a range of milk-based commodities. The LC-MS/MS method was compared to AOAC method 2010.01, based on affinity liquid chromatography, with and without the casein precipitation step. Heat stability of bovine IgG in a model nutritional product was evaluated.</p><p><strong>Method: </strong>After dilution in buffer, reduction, alkylation, and tryptic digestion, a single IgG1 + 3 isoform-specific peptide was used for quantitation of total IgG. The corresponding isotopically labeled peptide was used for internal standardization. Colostrum powder was used as matrix-matched calibrant.</p><p><strong>Results: </strong>Linearity was demonstrated in the 5-500 mg/L total IgG range. Within-day and intermediate precision were <4% and <6%, respectively, in two milk-based matrices and recovery was 95-99%. Major differences were noted between the LC-MS/MS method and AOAC method 2010.01 with and without casein precipitation, caused by factors such as loss of IgG during casein precipitation and inclusion of partially denatured IgG in the result. Heat processing loss of bovine IgG in a nutritional product was ca. 25% by the LC-MS/MS method.</p><p><strong>Conclusions: </strong>The LC-MS/MS method is applicable to milk-based commodities and nutritional products as well. A higher fraction of IgG (including denatured IgG) is accessible to measurement by LC-MS/MS than by AOAC method 2010.01. Casein precipitation in whey-based matrices causes loss of IgG.</p><p><strong>Highlights: </strong>A robust and widely applicable LC-MS/MS method using colostrum powder as calibrant and a single peptide marker for total IgG is being proposed.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145254360","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":"Elevated 7-Hydroxymitragynine Levels Found in Products Misbranded as Kratom.","authors":"Paula N Brown, Michael Chan, Xiaohui Zhang, Thomas Brendler","doi":"10.1093/jaoacint/qsaf094","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf094","url":null,"abstract":"<p><strong>Background: </strong>The leaves of Mitragyna speciosa, commonly called kratom, have been consumed for centuries for their energizing and analgesic properties. Kratom has gained popularity in North America with an estimated 16 million American consumers. Recently, a new product has emerged, labelled \"kratom extract\" but purportedly comprised of high concentrations of 7-hydroxymitragynine, an oxidation product of mitragynine, the primary alkaloid found in kratom.</p><p><strong>Objective: </strong>7-hydroxymitragynine, a potent mu-agonist, may pose a serious public health risk. Commercial products labelled as containing 7-hydroxymitragynine were evaluated to determine whether the constituent was present and in what concentration.</p><p><strong>Method: </strong>The concentrations of 7-hydroxymitragynine and mitragynine were determined by a previously validated HPLC-DAD method, AOAC 2017.14. The chromatographic profiles of products were also compared to authentic kratom leaf.</p><p><strong>Results: </strong>All products were described on their label as containing \"kratom extracts.\" The concentration of mitragynine ranged from 2.0-6.0 mg/g dry weight, substantially lower than typically found in kratom leaves or extracts. 7-hydroxymitragynine was determined at 22-75 mg/g dry weight, 5-28% higher than label claim. The chromatographic profiles of all products were inconsistent with kratom leaf, having exceptional levels of 7-hydroxymitragynine, lacking the other major naturally occurring alkaloids, and exhibiting peaks not observed in leaf.</p><p><strong>Conclusions: </strong>The products were verified to contain very high levels of 7-hydroxymitragynine, achievable only by synthetic means. These products differ significantly from authentic native kratom leaf and are not kratom extracts, as labelled, but rather synthetic derivatives that constitute unapproved new drugs being unlawfully marketed. There is an urgent need to differentiate kratom, which has been safely consumed for centuries, from 7-hydroxymitragynine that may pose a significant risk to public health.</p><p><strong>Highlights: </strong>Traditional kratom leaf is distinguished from products found to contain high concentrations of 7-hydroxymitragynine, only achievable by synthetic means. These unapproved new drugs contained other unidentified constituents, also not present in the native kratom leaf. The presence of concentrated 7-hydroxymitragynine products in the market masquerading as kratom is misleading, unlawful, and potentially dangerous to the public.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145254292","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":"Simultaneous and Quantitative Determination of Pyrrolizidine and Tropane Alkaloids in Food by LC-MS/MS, First Action 2025.02.","authors":"Thomas Bessaire, Claudia Mujahid, Pascal Mottier, Ke Du, Andrew Savage, Xun Fu, Leland Hunter, Charmaine Teo, Wai-Chinn Chan","doi":"10.1093/jaoacint/qsaf097","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf097","url":null,"abstract":"<p><strong>Background: </strong>Pyrrolizidine alkaloids (PAs) are natural toxins produced by > 6,000 flowering plant species. Some PAs are hepatotoxic and genotoxic carcinogens. Reported cases of direct poisoning in humans have already been documented, but the highest risk of human exposure occurs through the consumption of crops contaminated with PA-producing weeds. The European Commission Regulation EU 2023/915 set maximum levels (MLs) for the sum of 35 PAs in tea, herbal teas, herbs, and spice seeds at levels ranging from 75 to 1000 µg/kg (dry matter basis).</p><p><strong>Objective: </strong>The study describes the performance characteristics of a quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of the 35 PAs mentioned in AOAC INTERNATIONAL Standard Method Performance Requirements (SMPRs®) 2023.002 in tea, herbal infusions, herbs, seed spices, cereals, and honey. The analyte scope also comprised of nine additional PAs along with two tropane alkaloids (TAs, namely atropine and scopolamine) for which MLs are also set in plant-based products in European Union (EU 2023/915).</p><p><strong>Method: </strong>The protocol involves an extraction of toxins from food using aqueous sulfuric acid followed by solid-phase extraction purification. After elution and evaporation to dryness, the residue is reconstituted in water-methanol solution prior to LC-MS/MS analysis. Alkaline mobile phases are used to favor the separation of isomers. Identification of the analytes is conducted according to the confirmation criteria defined in EU 2023/2783 and SANTE 11312/2021v2 documents. Quantification is performed by isotopic dilution using four isotopically labelled PAs and two isotopically labelled TAs as internal standards.</p><p><strong>Results: </strong>Method validation workload was shared by three different laboratories on representative matrices from the following commodities: dried herbs, dried tea, dried herbal infusions, seed spices, cereals, and honey. The 35 target analytes specifically listed in the AOAC SMPR 2023.002 have been successfully validated (recovery 70-120%, repeatability < 20%, intermediate reproducibility < 20%), allowing LOQs down to 1 µg/kg.</p><p><strong>Conclusions: </strong>The performances of the method are in agreement with criteria defined in AOAC SMPR 2023.002.</p><p><strong>Highlights: </strong>The Expert Review Panel for Pyrrolizidine Alkaloids approved the present method as AOAC Official First Action 2025.02.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145254339","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":"Psychedelic Plants & Fungi: Regulatory Landscape, Analytical Challenges, and Quality Considerations.","authors":"Sharon L Brunelle","doi":"10.1093/jaoacint/qsaf093","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf093","url":null,"abstract":"","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Food Adulteration by Granular Formulation of Carbamates for Animal Poisoning: Chromatographic Determination and Vibrational Microspectroscopy.","authors":"Fernanda Ziegler Reginato, Karol Andriely de Vargas Paier, Maria Odete da Silva Dalan, Gustavo Andrade Ugalde, Sailer Santos Dos Santos, Marcelo Barcellos da Rosa, André Valle de Bairros","doi":"10.1093/jaoacint/qsaf092","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf092","url":null,"abstract":"<p><strong>Background: </strong>Carbamates have been used in intentional poisoning cases to cause death in both humans and animals. Granulated material containing carbamate pesticides is often applied to the victim's food, water, or drink. Toxicological analysis is essential to confirm such cases.</p><p><strong>Objective: </strong>This study aims to identify and quantify carbamates (methomyl, aldicarb, propoxur, carbofuran, and carbaryl) present in granular formulations used to adulterate food, through the application of chromatographic and vibrational microspectroscopic techniques.</p><p><strong>Methods: </strong>Thin layer chromatography (TLC), liquid chromatography with diode array detector (LC-DAD), gas chromatography-mass spectrometry (GC-MS), Attenuated total reflectance Fourier transform infrared (ATR-FTIR), and Raman microspectroscopy were performed. An aliquot of granular formulation was manually separated for chromatographic methods; dilutions were employed as test material treatment, while vibrational microspectroscopy techniques used raw material for toxicological analysis. A material similar to a shredded sausage containing suspected granular formulation was evaluated in chromatographic tests, and carbofuran was determined.</p><p><strong>Results: </strong>The proposed TLC methodology for identifying carbamates proved unfeasible due to insufficient chromatographic separation. The LC-DAD method was fully validated with success for methomyl, aldicarb, propoxur, carbofuran, and carbaryl, revealing a concentration of 744 µg/g of carbofuran in the suspected material, which was further confirmed by GC-MS. Raman microspectroscopy was not precise enough for carbamate identification, but ATR-FTIR demonstrated high specificity and robustness in identifying carbamate pesticides present in the test material.</p><p><strong>Conclusion: </strong>This study allowed determining the profile of carbamate pesticides in granular material when present in adulterated food/feed. To isolate the granules manually reduces the carrying of matrix interferents from food and/or feed. Furthermore, the techniques applied require only the raw material or its diluted versions, avoiding steps, high consumption of organic solvents, and reducing analysis time and cost.</p><p><strong>Highlights: </strong>LC-DAD was successfully validated for multiple carbamates. Minimal test material prep reduces solvent use, analysis time, and costs.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Determination and Confirmation of Psychedelic and Psychoactive Compounds in Botanical Materials by UPLC-ESI-MS/MS: Single Laboratory Validation.","authors":"Torey French, Anthony J Fontana, Robert LaBudde, Sharon L Brunelle, Sidney Sudberg","doi":"10.1093/jaoacint/qsaf081","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf081","url":null,"abstract":"<p><strong>Background: </strong>A multi-analyte method was developed for determination and confirmation of 26 natural and synthetic psychedelic and psychoactive compounds from botanical sources using a simple extraction procedure and UPLC-ESI-MS/MS analysis.</p><p><strong>Objective: </strong>The objective of this work was to develop and validate the method for analysis of psychedelic and psychoactive compounds in botanicals (fungi, cactus, leaf), extract (brewed tea), synthetic and simulated finished product (using maltodextrin).</p><p><strong>Methods: </strong>The method uses an acidified methanol solution for extraction and diphenhydramine as internal standard. Extracts are subjected to UPLC with tandem mass spectrometry using a phenyl-hexyl column and acidified mobile phase with acetonitrile gradient. Validation studies followed US Pharmacopeia <1225> and Official Methods of Analysis  SM Appendix K guidelines to assess specificity, accuracy (recovery), repeatability and intermediate precision, limits of detection and quantification, calibration curve linearity, system suitability, and robustness.</p><p><strong>Results: </strong>Average recovery of spiked replicates in matrix and surrogate matrixes varied between 88-124% for analytes in fungi, cactus, and leaf matrixes; 81-106% in brewed tea matrix; and 81-174% across all 26 analytes in maltodextrin. Norbaeocystin and harmaline were the only analytes outside the acceptance criterion (79-126%) in maltodextrin. The 90% upper confidence limit on repeatability was ≤33% for all analytes and matrixes except for norbaeocystin. Repeatability precision for analysis of matrixes with native analytes met the acceptance criterion of ≤ 33% in all cases and the 90% upper confidence limit on intermediate precision was ≤35% in all cases. LOQ values fell below the lowest calibration standard (0.016 µg/mL) for all 26 analytes.</p><p><strong>Conclusion: </strong>The method met the pre-determined acceptance criteria for recovery and precision in nearly all cases. Calibration curve linearity and system suitability measures were established.</p><p><strong>Highlights: </strong>A method for 26 psychedelic/psychoactive compounds was validated for analysis of botanical (fungi, cactus, leaf), extract (brewed tea), synthetic and simulated finished product (using maltodextrin) using native and surrogate botanicals.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145180739","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":"Plastic Additives in NIST Standard Reference Material 2585 and Settled Dust from Indoor Domestic and Working Environments.","authors":"Andrea Fricano, Patrizia Di Filippo, Donatella Pomata, Carmela Riccardi, Fabio Candiano, Giulia Simonetti, Francesca Buiarelli","doi":"10.1093/jaoacint/qsaf091","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf091","url":null,"abstract":"<p><strong>Background: </strong>Micro and nano plastics from textiles, rubber, cleaning products, etc. present in living and working environments can release toxic compounds, added to plastics to improve their properties.</p><p><strong>Objective: </strong>This study explores the presence of plastic additives in settled dust from domestic and workplace indoor environments.</p><p><strong>Methods: </strong>A Standard Reference Material was used to verify the validity of analytical method. Settled dust from two occupational environments (a sail loft and a tire shop), and from the home of a worker in the sail loft, is extracted, obtaining two fractions to be injected in GC-MS and in HPLC-MS/MS to detect and quantify 32 plastic additives. Results from working environments were also compared with those from a treating plant of waste electrical and electronic equipment (WEEE).</p><p><strong>Results: </strong>After a clean-up procedure and the use of matrix-matched calibration curves, the method proved to be reliable.Significant differences among the concentrations of analytes extracted from the workplace settled dust were not found except for higher values of Bis-2-ethylhexyl adipate (DEHA) and Bis-2-ethylhexyl phthalate (DEHP) in the tire shop and of dimethyl phthalate (DMP) and Diisodecyl phthalate (DiDP) in the sail loft. Comparing the results from house and work environments, higher concentrations of additives were found at home. The results from the present workplaces compared with those from a WEEE treating plant showed that the latter was a much more polluted environment.</p><p><strong>Conclusion: </strong>These preliminary results about the presence of plastic additives in the settled dust from living and working environments suggest that WEEE treating plant deserves more attention than others. Furthermore, the home environment hides some dangers for the presence of material that can release toxic compounds.</p><p><strong>Highlights: </strong>Optimization of a method for detecting plastic additives in environmental settled dust.NIST SRM 2585 analysis of phthalates, adipates, phosphates, citrates, trimellitate, benzoate, sebacate, dicarboxylate, benzo-triazole, -triazine.Exposure to plastic additives occurs both in workplace and living environments.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145093072","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":"Comparative Analysis of Volatile Constituents in Different Parts and Essential Oil of Pogostemon Cablin Using GC-MS Combined with Chemometrics.","authors":"Cheng Wang, Liang Hong, Weitong Gong, Qingwen Zhang, Shaoping Li, Jing Zhao","doi":"10.1093/jaoacint/qsaf090","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf090","url":null,"abstract":"<p><strong>Background: </strong>Pogostemon cablin (P. cablin) is a valuable medicinal plant used in traditional medicine and the fragrance industry, but quality control is challenging due to inconsistent stem-to-leaf ratios and frequent essential oil adulteration.</p><p><strong>Objective: </strong>This study compares volatile components in different parts (aerial parts, stems, leaves) and essential oil of P. cablin to support quality control (not less than 20% leaf) and its rational use.</p><p><strong>Methods: </strong>Volatile components in 21 batches of aerial parts, stems, leaves, and 13 batches of essential oils were analyzed using GC-MS. Multivariate curve resolution-alternating least squares (MCR-ALS) was used for resolving co-eluted peaks, and chemical fingerprinting with chemometric techniques like hierarchical cluster analysis (HCA), principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and orthogonal partial least squares discrimination analysis (OPLS-DA) was applied.</p><p><strong>Results: </strong>Volatile profiling identified 56, 47, 28, and 45 components in the aerial parts, leaves, stems, and essential oil of P. cablin, respectively. MCR-ALS resolved ten major volatile compounds to create chemical fingerprints for each analytical sample type. Quantitative analysis showed higher patchouli alcohol in leaves (12.47 mg/g) compared to stems (2.05 mg/g), while stems had more pogostone (2.71 mg/g vs. 1.40 mg/g in leaves). Aerial parts and essential oil showed significant compositional differences. Based on the results of qualitative and quantitative analysis, chemometric methods, including HCA, PCA, PLS-DA and OPLS-DA, clearly differentiated the four types of P. cablin analytical samples.</p><p><strong>Conclusions: </strong>Significant differences in volatile components across P. cablin parts and its essential oil support quality control (not less than 20% leaf) and rational use.</p><p><strong>Highlights: </strong>This study is the first to use MCR-ALS and other chemometrics for qualitative and quantitative analysis of P. cablin parts and essential oils, aiding quality control.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145093013","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":"Development of ELISA for Cyclopiazonic Acid in Various Foods and Its Validation Study Using LC/UV and LC/TOF-MS.","authors":"Koichi Saito, Ayako Kitora, Kana Hekizono, Maimi Kurata, Kazue Banba, Rie Ito, Hiroshi Akiyama","doi":"10.1093/jaoacint/qsaf088","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf088","url":null,"abstract":"<p><strong>Background: </strong>Food contamination with mycotoxins has become a concern in recent years. Cyclopiazonic acid (CPA) is a mycotoxin found in foods, such as peanuts, corn, and cheese, which poses health hazards to humans.</p><p><strong>Objective: </strong>To develop and validate an enzyme-linked immunosorbent assay (ELISA) for detecting CPA in various food matrices using appropriate pretreatment methods, with liquid chromatography/ultraviolet detection (LC/UV) and/or LC/time of flight mass spectrometry (TOF-MS).</p><p><strong>Methods: </strong>Pretreatment of food was optimized using liquid-liquid extraction and solid-phase dispersive extraction (SPDE), and validated across liquid and solid foods. Indirect competitive ELISA was performed. The data from recovery experiments were analyzed using a one-way analysis of variance.</p><p><strong>Results: </strong>All food specimens added at high concentrations yielded satisfactory ELISA results and exhibited a strong correlation with the LC/UV measurements. In the case of moderate-concentration additions, satisfactory accuracy was obtained for foods other than spices. At low concentrations, the results of ELISA and instrumental analyses (LC/UV and LC-TOF/MS) were comparable for sake, soy sauce, noodle soup, corn, and miso. However, elevated values were observed for the ELISA of cheese, peanuts, and spices.</p><p><strong>Conclusion: </strong>Although the low recovery levels of CPA in spices suggests the need for a more effective cleanup method, the developed ELISA is suitable as a rapid screening method for CPA in various foods, with LC confirmation recommended in complex matrices.</p><p><strong>Highlights: </strong>The use of SPDE for CPA analyses resulted in effective cleanup, which subsequently enabled the application of ELISA for screening.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145093047","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 HPLC-MS-Based Method for Determination of the D- and L- 5-Methytetrahydrofolate Isomer Ratio in Dietary Supplements.","authors":"Xiaojuan Wang, Deepesh Pandey, Suma Vavilala, Laura Oh, Pavel Gusev, Karen W Andrews, Pamela Pehrsson, James M Harnly, Jianghao Sun","doi":"10.1093/jaoacint/qsaf089","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf089","url":null,"abstract":"<p><strong>Background: </strong>5-Methytetrahydrofolate (5-MTHF), a reduced form of folate (vitamin B9) marketed as a potentially more bioavailable alternative to folic acid, is a key ingredient in dietary supplement (DS), particularly prenatal multivitamin/minerals. Its synthesis may produce racemic mixtures of bioactive L-5-MTHF and inactive D-5-MTHF, raising concerns about labeling accuracy, integrity, and health impacts. DS manufacturers consider D-5-MTHF an impurity and do not use racemic mixtures of 5-MTHF in their products.</p><p><strong>Objective: </strong>This study aimed to develop and validate an HPLC-MS-based method to separate and quantify the D and L-5-MTHF diastereomers in various DS matrices and dosage forms.</p><p><strong>Methods: </strong>A single-laboratory validated method was developed for determination of 5-MTHF isomers in different dosage forms of DS using targeted selected ion monitoring (Targeted SIM) with a ChiralPak HSA column. L-cysteine was selected as an extraction stabilizer, and the mobile phase was composed of 100 mM ammonium acetate and 1% v/v acetic acid in acetonitrile (ACN) at a 97:3 ratio.</p><p><strong>Results: </strong>The HPLC/MS method was able to separate D-and L-5-MTHF diastereomers by isocratic elution within 20 min. It showed great linearity with the concentration range of 1 µg/mL to 160 µg/mL of 5-MTHF with R  2>0.99. The method was validated for the limit of detection (LOD), limit of quantitation (LOQ), linear range, specificity, and recovery according to the guidelines described Q2(R1) Validation of Analytical Procedures: Text and Methodology Guidance for Industry. The method was then applied for the analysis of 101 samples, and two samples were found to exceed the USP limit of ≤ 1% D-5-MTHF relative to L-5-MTHF (1.39% and 50.28%).</p><p><strong>Conclusion: </strong>This performance of HPLC-MS method established effectively determines D- and L-5-MTHF ratios in complex DS, serving as a reliable tool to detect adulteration with racemic mixtures in prenatal and other 5-MTHF-labeled containing DS. It is significantly faster than previously reported method and possesses great selectivity in separation of the diastereomers in different form of DS.</p><p><strong>Highlights: </strong>Successfully validated an HPLC-MS method for quantifying D- and L-5-MTHF diastereomers in diverse DS dosage forms, aiding enantiomeric purity assessment.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145093008","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}