Journal of AOAC International最新文献

{"title":"Development of Blocking ELISA Kit Based on Recombinant Monoclonal Antibody for the Serological Diagnosis of Peste Des Petits Ruminants Virus.","authors":"Chunyan Feng, Runze Li, Caixia Wang, Yujing Geng, Bin Wu, Haoyang Yu, Haoxuan Li, Zhou Zhang, Zhen Yang, Shaoqiang Wu, XiangMei Lin","doi":"10.1093/jaoacint/qsaf077","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf077","url":null,"abstract":"<p><strong>Background: </strong>Peste des petits ruminants (PPR) was a highly contagious and economically devastating disease affecting sheep and goats. The performance and cost of serological diagnostic kit remained a major challenge for PPR eradication.</p><p><strong>Objective: </strong>The nucleocapsid (N) protein of PPR virus and its monoclonal antibody (mAb) were engineered for development of a blocking ELISA.</p><p><strong>Methods: </strong>The N protein was truncated and expressed by eukaryotic expression system. The mAb 65A8 was recovered, followed by sequencing and cloning of its variable regions into expression vector. The recombinant mAb was expressed in 293F cells. Key characteristics of the recombinant mAb were evaluated. A blocking ELISA was developed and validated. Comparison between the blocking ELISA and a commercial blocking ELISA kit was performed.</p><p><strong>Results: </strong>The truncated N protein exhibited a higher yield compared to the full-length N protein. Recombinant mAb 65A8 demonstrated the same blocking efficacy as that of ascites-derived mAb. The yield of recombinant mAb reached 5.3 mg/100 mL. The affinity constant of recombinant mAb 65A8 was comparable to that of the ascites -derived 65A8 (3.48 × 109 L/mol vs 6.76 × 109 L/mol). Based on the truncated N and recombinant mAb, a blocking ELISA was established. The detection limit of blocking ELISA was 1:128 and 1:32 based on strong positive serum and weak positive serum, respectively. Inter- and intra-assay coefficients of variation were <10%, indicating robust reproducibility. The blocking ELISA kit showed high coincidence with the commercial ELISA kit recommended by World Organization for Animal Health reference lab.</p><p><strong>Conclusion: </strong>This study had successfully developed a reliable and cost-effective blocking ELISA by protein engineering of the antigen and mAb.</p><p><strong>Highlights: </strong>The mAb of PPR Virus was recombinantly expressed in 239F cells, which was employed in development of a reliable blocking ELISA. This work provided novel ideas and methods for the developing of the blocking ELISA kits.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144983966","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":"Modification of the D3 Array Combined (Formerly the DetectX Combined) for the Detection of Aspergillus, Salmonella, and Shiga Toxin-Producing Escherichia coli in Dried Cannabis Flower with Optional Enrichment: AOAC Performance Tested MethodSM 012201.","authors":"Meghan Hottinger, Mike Leasia, Mark Schwartz, Benjamin A Katchman","doi":"10.1093/jaoacint/qsaf080","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf080","url":null,"abstract":"<p><strong>Background: </strong>The D3 Array Combined assay (PTM 012201) is a qualitative test for the detection of Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus terreus, Salmonella species, and a broad range of Shiga toxin-producing Escherichia coli (STEC) serogroups in cannabis matrixes. The test involves extraction of nucleic acids from samples followed by preamplification, Loci polymerase chain reaction (PCR), and subsequent Labeling PCR. The PCR amplification product is then hybridized to a DNA microarray. Detection of bacterial and fungal pathogens is determined by fluorescence.</p><p><strong>Objective: </strong>The objective of this study was to validate the D3 Array Combined method with an optional enrichment step in dried cannabis flower (delta 9-tetrahydrocannabinol (THC) >0.3%), with the use of the Live/Dead Enrichment (LDE) reagent. The method modification was validated with optional enrichment time points at 0, 2, and 24 hours.</p><p><strong>Methods: </strong>The evaluation followed study designs from the AOAC Microbiology guidelines and AOAC Standard Method Performance Requirements (SMPRs®) 2019.001, 2020.02, and 2020.012. The modified method was compared to SMPR confirmation procedures and an unpaired Aspergillus consensus method. The results were evaluated utilizing a probability of detection (POD) statistical model.</p><p><strong>Results: </strong>Results showed no statistically significant difference, at the 95% and 90% confidence intervals, between the candidate method presumptive D3 Array Combined positive results and the confirmed cultural positive results at all contamination levels and enrichment time points.</p><p><strong>Conclusions: </strong>This study provides data that demonstrate the PathogenDx D3 Array Combined is a reliable method for detection of Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus terreus, Salmonella species, and a broad range of STEC serogroups in dried cannabis flower from 0-24 hours enrichment, for all these organisms.</p><p><strong>Highlights: </strong>This highly sensitive method provides results within hours of sample acquisition, compared to qPCR or cultural methods where results are obtained in 2-7 days. This method also provides the option of enrichment, providing compliance options for all cannabis testing state regulations.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144984113","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":"High-Throughput Quantification of Pesticide Residues in Complex Matrix (Chilli Powder) Using Liquid Chromatography Tandem Mass Spectrometry: Inter- and Intra-Day Validation.","authors":"Raviraj Chandrakant Shinde, Macky Suraliwala, Dharmendra Kumar, Sagar Atugade, Pandit Shiragave","doi":"10.1093/jaoacint/qsaf079","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf079","url":null,"abstract":"<p><strong>Background: </strong>Chilli powder is a widely consumed spice, however during cultivation of chilli crops are often subjected to pesticide treatments to control pests and diseases. Thus, monitoring pesticide residues in such matrices is crucial for food safety and to comply with national as well as international regulations for export/import purposes. However, accurate analysis of pesticide residue in chilli powder is challenging due to its complex nature.</p><p><strong>Objective: </strong>To develop and validate a high-throughput LC-MS/MS method for the quantification of multiclass pesticide residues in a complex chilli powder matrix.</p><p><strong>Methods: </strong>The acetonitrile-based extraction method was optimized for chilli powder samples. The targeted analytes were separated using reverse-phase liquid chromatography and detected using tandem mass spectrometry. Validation was conducted following SANTE guideline to comply with regulatory requirements.</p><p><strong>Results: </strong>The method with an optimised sample preparation workflow demonstrated a lower matrix effect of < 35% for the target pesticides. The LOQ was determined to be 0.005 mg/kg for 135 analytes, with recovery ranging from 70 to 110%, and intra-day and inter-day precision (%RSD) were below 15%. Analysis of market/incurred samples and measurement uncertainty further provided more confidence on the method performance.</p><p><strong>Conclusions: </strong>The developed LC-MS/MS method provides a robust, sensitive, and high-throughput approach for the quantification of pesticide residues in complex chilli powder. Its intra- and inter-day validation confirms suitability for routine analysis in food safety laboratories.</p><p><strong>Highlights: </strong>A high-throughput LC-MS/MS method is developed for pesticide analysis in complex chilli powder. The method was validated according to SANTE 11312/2021-v2 with excellent precision and accuracy. Suitable for routine food safety monitoring of wide range of pesticide residues in a testing laboratory.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144984023","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":"Multivariate Modeling-Enhanced Stable Isotopic Origin Traceability of Qinghai-Tibet Plateau Rape Honey.","authors":"Bin Li, Guigong Geng, Luqiong Miao, Xianxian Mei, Jialu Zhou, Yuyan Fei, Rui Zou, Zhi Liu, Dongfeng Yang","doi":"10.1093/jaoacint/qsaf076","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf076","url":null,"abstract":"<p><strong>Rationale: </strong>Qinghai-Tibet Plateau (QTP) rape honey, recognized as a Protected Geographical Indication (PGI) product in China, has faced significant challenges due to fraudulent mislabeling of its origins in the market. To ensure the authenticity of PGI honey products and uphold market integrity, it is crucial to develop a rapid, precise, and efficient geographical traceability technology.</p><p><strong>Methods: </strong>A total of 208 honey samples were collected from QTP (n = 71) and 5 provinces in the southern region (SR, n = 137) of China. Stable isotope ratios (δ13C, δ15N, δ2H, and δ18O) of bulk honey, endogenous proteins, and saccharides (glucose, fructose, and sucrose) were measured. One-way analysis of variance (ANOVA) was employed to analyze regional differences among the variables. Partial least squares discriminant analysis (PLS-DA) and linear discriminant analysis (LDA) models were constructed based on stable isotopic data to discriminate honey sample origins.</p><p><strong>Results: </strong>ANOVA indicated the geospatial differences (P < 0.05) in δ2H and δ18O of bulk honey, as well as all four ratios of honey protein, are significant between QTP and SR. LDA exhibited superior discrimination performance, with leave-one-out cross-validation accuracies of 87.3% for QTP, 89.1% for SR.</p><p><strong>Conclusion: </strong>An integrated strategy combining stable isotope ratios analysis with multivariate modeling provides an accurate and effective verification method for geographical origin traceability of high-value honey from QTP. This approach provides a reliable tool to address the issue of fraudulent mislabeling of PGI rape honey.</p><p><strong>Highlights: </strong>Stable isotopic signatures of Qinghai-Tibet Plateau rape honey were discussed. Bulk and component-specific isotopic ratios were informative geospatial indicators. Machine learning algorithms significantly enhanced honey origin discrimination. LDA accuracy for Qinghai-Tibet Plateau honey samples reached up to 87.3%. This strategy was developed to combat origin mislabeling and ensure food integrity.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144984056","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":"Opportunities to Fill Gaps in Reference Methods for Vitamins in Foods and Dietary Supplements.","authors":"Erik J M Konings, Philippe J Eugster, Jinchuan Yang, Shulin Feng, Kai Liu, Elaine Jobgen, Eystein Oveland","doi":"10.1093/jaoacint/qsaf074","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf074","url":null,"abstract":"<p><strong>Background: </strong>A recent review concluded that most current international reference methods to accurately determine vitamin content in foods and food supplements are no longer fit-for-purpose. Most reference methods originate from the 1990's and science, technology, and statutory regulations have changed considerably since.</p><p><strong>Objective: </strong>AOAC INTERNATIONAL hosted a symposium during its annual meeting in Baltimore in August 2024 to highlight and discuss method gaps and share cutting-edge technology/methods for future consideration as reference methods for vitamins in foods and dietary supplements.</p><p><strong>Methods: </strong>Five invited speakers discussed gaps, opportunities and priorities when it comes to future reference method needs in the area of vitamins in foods and dietary supplements.</p><p><strong>Results: </strong>Speakers successively advocated for: 1. Harmonization of analytical methods which is closely related to the harmonization of definitions on what compounds to include and the expression of results with a focus on retinol and retinyl esters as an example. 2. At present there is no way to verify or validate the type of vitamin E on nutrition labels. Chiral analysis is needed to differentiate the type of vitamin E between natural and synthetic, which is needed to express the amount of bioactive vitamin E. 3. With the objective to develop a cost- and time-efficient method that is accurate and able to quantify all B-vitamins, a LC-ECI-MS/MS method for the simultaneous determination of six B-vitamins: B1, B2, B3, B5, B6 and B7 in foods was presented as potential new reference method. 4. Attention was requested for the different types of ingredient manufacturing processes, which can cause significant challenges for accurate and precise testing of vitamins.</p><p><strong>Conclusion: </strong>Opportunities discussed gave a good perspective of future priorities to be considered when establishing new reference methods for the determination of vitamins in foods and dietary supplements.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144877618","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":"Towards Standardized Products Containing Biomass of Psilocybe cubensis Fungi.","authors":"Kimberley Foster, Isaac Morrison, Shemar Daniel, Johann Antoine, Babumon Thankappan, Winston De La Haye, Marshall Tyler, Charles Grant, Rupika Delgoda","doi":"10.1093/jaoacint/qsaf072","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf072","url":null,"abstract":"<p><strong>Background: </strong>The consumption of dried fruiting bodies of Psilocybe cubensis can be traced over centuries, guided by Mesoamerican curanderas, Western medical practitioners, and fungal enthusiasts, all seeking mental wellbeing. There's a notable resurgence in interest both in the fungal biomass and psilocybin, the psychoactive tryptamine, albeit the global regulatory restrictions, following enlistment in the UN convention on psychotropic substances.</p><p><strong>Objectives: </strong>To evaluate consistency in psilocybin potency and to determine levels of microbial, pesticidal and heavy metal content, in products encompassing biomass of uniformly cultivated P. cubensis.</p><p><strong>Methods: </strong>In a legally sanctioned, unique lab in Jamaica, we cultivated P. cubensis according to published methods, then dried, pulverized, extracted and tested fruiting bodies for tryptamine content using an Agilent HPLC 1290 Infinity assembly. Colony counting was employed for E. coli, yeast, mold, coliform presence, while a Neogen's Veratox® ELISA assay assessed mycotoxin content. Agilent GCMS and LC assemblies evaluated for pesticidal content while heavy metals (As, Cd, Pb, Hg) were determined using instrumental neutron activation analysis (INAA), energy-dispersive X-ray fluorescence (ED-XRF, and direct mercury analysis (DMA) by thermal decomposition-amalgamation-atomic absorption spectrometry (TDA-AAS), respectively.</p><p><strong>Results: </strong>Mean psilocybin and psilocin content in dried cultivated P. cubensis was 1.14 ± 0.17% by weight, however there was batch variability, potentiating significant differences in projected dosage, particularly for and above 3 g. The homogenized biomass was deemed safe, with acceptable levels of microbial, mycotoxin, pesticidal, and heavy metal contents, and no significant carcinogenic or other health hazards. Encapsulated biomass stably maintained tryptamine content for 11 months.</p><p><strong>Conclusion: </strong>Standardized, safe biomass suitable for human consumption can be achieved using P. cubensis cultivated under stringent, aseptic conditions. Given the observed variability, it is highly recommended that each batch is tested for tryptamine content. Our results may be useful for policy makers, cultivators, clinicians and consumers.</p><p><strong>Highlight: </strong>The present study provides a basis for regular potency testing of P. cubensis biomass and substantiates their potential use in clinical trials as a high quality, standardized and safe product.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144850184","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":"Raman Spectroscopy Coupled with Chemometric Techniques for Authenticity Assessment of Camel Milk Powder.","authors":"Omar Ait El Alia, Abdennacer El Mrabet, Soumaya Boukrouh, Morad Kaddouri, Khalid Boutoial, Aimen El Orche","doi":"10.1093/jaoacint/qsaf075","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf075","url":null,"abstract":"<p><strong>Background: </strong>Adulteration of camel milk powder with cheaper alternatives such as cow milk powder has become a growing concern, compromising both nutritional quality and consumer trust. Detecting such adulteration is critical for ensuring product authenticity, especially given the rising demand for camel milk in niche health markets.</p><p><strong>Objective: </strong>This study explores the application of Raman spectroscopy in conjunction with chemometric techniques for the detection and quantification of adulteration in camel milk powder with cow milk powder.</p><p><strong>Method: </strong>Camel milk powder was adulterated with cow milk powder across a range of concentrations from 0% to 50%. Raman spectra of these mixtures were analyzed using PCA for dimensionality reduction, followed by PLSR modeling with different spectral pretreatments (raw, Savitzky-Golay, gap derivative). Interval PLS (IPLS) in backward mode was applied to enhance variable selection.</p><p><strong>Results: </strong>PCA captured 99.6% of spectral variance. The raw PLSR model already showed strong predictive power (R2cv = 95.93%). Savitzky-Golay further boosted performance (R2test = 99.47%), while the gap derivative achieved near-perfect prediction (R2test = 99.94%, RMSEtest = 1.10). IPLS modeling significantly improved robustness, yielding high accuracy (R2test = 98%) with fewer variables.</p><p><strong>Conclusion: </strong>The findings indicate that the integration of Raman spectroscopy with PCA, PLSR, and IPLS constitutes a robust, precise, and reliable approach for the detection of adulteration in camel milk powder.</p><p><strong>Highlights: </strong>The application of Raman spectroscopy coupled with chemometric modeling proves to be an efficient and robust analytical tool for quality control in the dairy industry, enabling the accurate detection of adulteration and ensuring the authenticity and safety of camel milk powder.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144850183","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":"Use of a Spectrophotometric Method for the Detection of Adulterants in Commercial Fulvic Acid Products.","authors":"Elena A Vialykh, Shelby Buckley, Julia Gentile, Fernando L Rosario-Ortiz, Richard T Lamar, Jarrod Psutka, Mohammad Rahbari","doi":"10.1093/jaoacint/qsaf073","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf073","url":null,"abstract":"<p><strong>Background: </strong>Numerous products manufactured from non-humic sources have flooded the market claiming to be fulvic acids. The challenge is finding an easy method to distinguish between products containing genuine fulvic fractions and those containing adulterants. UV spectrophotometry has been widely used to study the fulvic fraction extracted from humic substances, with multiple metrics derived from UV absorption spectra developed and implemented by researchers.</p><p><strong>Objective: </strong>Leverage ten indices that are characteristic features of the UV spectra of hydrophobic fulvic acids to differentiate products containing authentic fulvic fractions from those containing adulterants.</p><p><strong>Methods: </strong>Fulvic fractions were diluted to 5 ppm carbon and UV spectra were obtained. Spectra were normalized and analyzed to calculate 10 different indices. The percent difference between the index values of the product and the corresponding index values for the SRFA and PPFA standards were calculated. An equally weighted average for all 10 indices was calculated and a 70% cut-off value was used for the average percent error as a screening tool to distinguish products containing fulvic fractions from adulterants.</p><p><strong>Results: </strong>Fifty-four test samples were analyzed, with nine samples being analyzed by two different labs using the established method. Fourteen of the 25 commercial products studied were found to contain fulvic fractions. Increased metal ion concentration within the investigated range did not impact the average percent error calculated, nor did varying the total organic carbon concentrations of the test portions within the range of 1-10 ppm.</p><p><strong>Conclusion: </strong>The method investigated could be a suitable screening tool for most commercial products and is capable of accurately distinguishing products that contain fulvic fractions.</p><p><strong>Highlights: </strong>The method accurately found all 11 fulvic fractions isolated from known humic substances as fulvic, and all 11 test samples prepared from non-humified materials as non-fulvic.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144850185","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 an RP-HPLC Method for the Determination of Lidocaine Hydrochloride in Injectable Formulation: Combining White Analytical Chemistry and Experimental Design with Eco-Friendly and Cost-Effective Method.","authors":"Elif Özdemir, Şule Dinç-Zor","doi":"10.1093/jaoacint/qsaf071","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf071","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;In recent years, environmental impact, human health, and cost have become increasingly important in chromatographic analysis of pharmaceutical compounds. Traditional methods use organic solvents like acetonitrile (ACN) and methanol (MeOH), which are volatile, flammable, toxic, and environmentally harmful. By using the available comprehensive data on the effects of chemicals on human health and the environment, informed choices should be made about which chemicals are more suitable for a given synthesis or process, taking even a small step toward green chemistry.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Objective: &lt;/strong&gt;In this study, the aim is to develop a green high-performance liquid chromatography (HPLC) method for the analysis of lidocaine by replacing the toxic solvents traditionally used in the mobile phases of classical chromatographic methods with greener alternatives. To achieve this, ethanol is used as the organic modifier in the mobile phase without compromising analytical performance, thereby enabling a transition to green chromatography.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;The independent variables considered were the pH of the mobile phase, flow rate, and ethanol content in the mobile phase for the optimization step. Experimental runs were selected randomly, and a total of 15 experiments were conducted. Response parameters for each HPLC chromatogram were calculated, evaluated using regression analysis, and the accuracy of the results was tested using ANOVA.The Derringer desirable function was utilized to optimize the conditions. Accordingly, the optimal conditions determined were a mobile phase pH of 4.0, with a 1.3 mL/min flow rate and an ethanol content in the mobile phase of 25%.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The developed new green method offers an environmentally friendly, sensitive, and reliable alternative as lidocaine is determined using a high-performance liquid chromatography technique.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;The developed and validated green HPLC method can be proposed as an alternative to the conventional HPLC methods reported by the USP and other sources, which are not environmentally or human health-friendly, for the analysis of lidocaine in pharmaceutical preparations. The use of ethanol instead of potentially toxic organic solvents minimizes harm to both the environment and analyst health. Additionally, the method offers advantages such as reduced analysis time, solvent and time savings, and the absence of labor-intensive sample and solvent preparation steps, making it an attractive option. It is considered that the developed method could be particularly useful in the pharmaceutical industry, especially in quality control laboratories where rapid and high-throughput analyses are conducted, as well as in R&D studies.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Highlights: &lt;/strong&gt;A green and sustainable HPLC method for lidocaine analysis was developed and validated. Chromatographic optimization was achieved using Design of Expe","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144777421","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":"Validation of Enzytec™ Liquid Combi Lactose/D-Galactose for Enzymatic Determination of Lactose and D-Galactose in Selected Foods: Official Method 2024.10 First Action.","authors":"Markus Lacorn, Thomas Hektor","doi":"10.1093/jaoacint/qsaf070","DOIUrl":"https://doi.org/10.1093/jaoacint/qsaf070","url":null,"abstract":"<p><strong>Background: </strong>Lactose is a disaccharide composed of D-galactose and D-glucose. Lactose composes 2-8% of milk by weight. Lactose is digested in the small intestine by the enzyme lactase. Lactase deficiency can lead to lactose intolerance in adults.</p><p><strong>Objective: </strong>Enzytec™ Liquid Combi Lactose/D-Galactose is an enzymatic test kit for the determination of lactose and D-galactose in milk, milk products, infant formula, chocolate, sausage meat, almond milk, salad dressing, bread and cookies. Excluded are lactose-free dairy products that have been enzymatically treated with lactase.</p><p><strong>Methods: </strong>The kit contains two sets of reagents in a ready-to-use format and is suitable for automation. Lactose is cleaved with ß-galactosidase. Resulting D-galactose is oxidized by a galactose-dehydrogenase and NAD+. The amount of resulting NADH is equivalent to the converted amount of lactose and free D-galactose. For measurement of free D-galactose alone the second set of reagents is used.</p><p><strong>Results: </strong>Trueness of the lactose measurement was checked using certified reference materials with recoveries for milk, milk products, infant formula, and chocolate between 94 and 103%. Spiking experiments at low levels in almond milk, salad dressing, feta cheese, cookies, and bread revealed recoveries between 92 and 121%. Intermediate precision varied between 3.0 and 10.6% for lactose and between 6.5 and 7.8% for D-galactose. For automation, three applications with different test volumes were validated. Linearity is given from 12.5 mg/L to 12.5 g/L for lactose and 4.0 mg/L up to 6.25 g/L for galactose.</p><p><strong>Conclusions: </strong>The method is robust and accurate for manual and automated applications. The method was approved as AOAC Official Method of Analysis℠.</p><p><strong>Highlights: </strong>The ready-to-use components of the test kit have a shelf life of at least 24 months.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144710398","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}