{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T05:15:47Z","timestamp":1788671747229,"version":"build-2803163510"},"reference-count":41,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T00:00:00Z","timestamp":1788480000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004539","name":"Kasetsart University","doi-asserted-by":"crossref","award":["FF(KU)22.68"],"award-info":[{"award-number":["FF(KU)22.68"]}],"id":[{"id":"10.13039\/501100004539","id-type":"DOI","asserted-by":"crossref"}]},{"award":["FF(KU)22.68"],"award-info":[{"award-number":["FF(KU)22.68"]}],"id":[{"id":"https:\/\/ror.org\/05gzceg21","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Methods Protoc.","MPs"],"abstract":"<jats:p>Carnosine and anserine are bioactive, histidine-containing dipeptides recognized for their antioxidant, pH-buffering, and neuroprotective functions. However, their precise quantification is frequently compromised by their high structural similarity and complex matrix-induced analytical interferences. This study developed and validated an optimized, simplified extraction protocol coupled with high-performance liquid chromatography diode array detection (HPLC-DAD) for the simultaneous determination of both dipeptides across diverse meat matrices, including Thai native black-bone chicken (Nin-Kaset), commercial broiler, pork, beef, and buffalo meat. Extraction efficiency was systematically optimized by modulating the solution pH. Under optimized conditions, the validated method demonstrated excellent linearity (1\u2013200 \u00b5g mL\u22121, r2 \u2265 0.9990) with limits of detection and quantification (LOD\/LOQ) values at 0.0093\/0.0283 mg g\u22121 for carnosine and 0.0139\/0.0420 mg g\u22121 for anserine. Method accuracy was confirmed by satisfactory recoveries ranging from 96.64% to 102.68%, with repeatability values (RSDs) of 0.70\u201314.18%. Application to real matrices revealed distinct dipeptide profiles. Pork contained the highest carnosine level (3.59 \u00b1 0.19 mg g\u22121), whereas white-feather Nin-Kaset exhibited the highest anserine level (6.77 \u00b1 0.40 mg g\u22121). Overall, the validated framework offers a rapid, cost-effective, and robust approach for routine dipeptide profiling, supporting future applications in food authentication, nutritional evaluation and functional food development.<\/jats:p>","DOI":"10.3390\/mps9050131","type":"journal-article","created":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T16:07:32Z","timestamp":1788538052000},"page":"131","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Quantification of Carnosine and Anserine in Diverse Meat Matrices via Optimized Extraction and Validated HPLC-DAD"],"prefix":"10.3390","volume":"9","author":[{"given":"Chalita","family":"Kaewkhow","sequence":"first","affiliation":[{"name":"Department of Chemistry, Faculty of Science, Kasetsart University, Bangkok 10900, 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Meat Consumption Worldwide from 1990 to 2024, by Meat Type. Available online: https:\/\/www.statista.com\/statistics\/274522\/global-per-capita-consumption-of-meat\/."},{"key":"ref_2","unstructured":"U.S. Department of Agriculture, and USDA (2026, February 14). Chicken Meat Production Worldwide from 2012 to 2025. Available online: https:\/\/www.statista.com\/statistics\/237637\/production-of-poultry-meat-worldwide-since-1990\/?srsltid=AfmBOoqii4u-QUhNlseXokxTIKaRaIYlQNvrCKWdsm2cdkPyxrpmhx8N."},{"key":"ref_3","first-page":"218","article-title":"Functional Food Science and Bioactive Compounds","volume":"8","author":"Martirosyan","year":"2025","journal-title":"Bioact. Compd. Health Dis."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Andrade, M.A., Barbosa, C.H., Shah, M.A., Ahmad, N., Vilarinho, F., Khwaldia, K., Silva, A.S., and Ramos, F. (2022). Citrus By-Products: Valuable Source of Bioactive Compounds for Food Applications. Antioxidants, 12.","DOI":"10.3390\/antiox12010038"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"148054","DOI":"10.1016\/j.foodchem.2026.148054","article-title":"Efficient capillary electrophoretic analysis of anthraquinones and phenolic acids in functional foods using a novel deep eutectic solvent\u2013in\u2013deep eutectic solvent microemulsion","volume":"506","author":"Fan","year":"2026","journal-title":"Food Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"141675","DOI":"10.1016\/j.foodchem.2024.141675","article-title":"Targeted metabolite and molecular profiling of carotenoids in rose petals: A step forward towards functional food applications","volume":"464","author":"Cheng","year":"2025","journal-title":"Food Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s40550-025-00122-0","article-title":"Comparative analysis on plant-based formulated tiger nut (Cyperus esculuntus) yoghurts (prebiotic, probiotic and symbiotic) fortified with fresh moringa leaf tea as functional foods","volume":"13","author":"Yeboah","year":"2026","journal-title":"Food Saf. Risk"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"105607","DOI":"10.1016\/j.tifs.2026.105607","article-title":"Difructose anhydrides as emerging functional food ingredients: Production methods, therapeutic potentials, and analytical perspectives","volume":"170","author":"Sojan","year":"2026","journal-title":"Trends Food Sci. Technol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"144673","DOI":"10.1016\/j.foodchem.2025.144673","article-title":"Oat-grape beverages enriched with anthocyanins from jambolan (Syzygium cumini): A novel plant-based probiotic functional food","volume":"487","author":"Damasceno","year":"2025","journal-title":"Food Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1939","DOI":"10.1016\/j.foodchem.2010.12.036","article-title":"Determination of carnosine, anserine, homocarnosine, pentosidine and thiobarbituric acid reactive substances contents in meat from different animal species","volume":"126","author":"Peiretti","year":"2011","journal-title":"Food Chem."},{"key":"ref_11","first-page":"147","article-title":"Quantitative analysis of carnosine and anserine in foods by performing high performance liquid chromatography","volume":"26","author":"Mori","year":"2015","journal-title":"Biol. Trace Elem. Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Charoensin, S., Laopaiboon, B., Boonkum, W., Phetcharaburanin, J., Villareal, M.O., Isoda, H., and Duangjinda, M. (2021). Thai Native Chicken as a Potential Functional Meat Source Rich in Anserine, Anserine\/Carnosine, and Antioxidant Substances. Animals, 11.","DOI":"10.3390\/ani11030902"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1007\/s00210-024-03427-7","article-title":"A comprehensive review on physiological and biological activities of carnosine: Turning from preclinical facts to potential clinical applications","volume":"398","author":"Kumar","year":"2025","journal-title":"Naunyn-Schmiedeberg\u2019s Arch. Pharmacol."},{"key":"ref_14","first-page":"FC01","article-title":"Evaluation of Antidiabetic and Antioxidant Activities of L-carnosine using Enzyme Inhibition and Free Radical Scavenging Assays: An In-vitro Study","volume":"17","author":"Malathy","year":"2023","journal-title":"J. Clin. Diagn. Res."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Maugeri, S., Sibbitts, J., Privitera, A., Cardaci, V., Di Pietro, L., Leggio, L., Iraci, N., Lunte, S.M., and Caruso, G. (2023). The Anti-Cancer Activity of the Naturally Occurring Dipeptide Carnosine: Potential for Breast Cancer. Cells, 12.","DOI":"10.3390\/cells12222592"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Caruso, G., Godos, J., Castellano, S., Micek, A., Murabito, P., Galvano, F., Ferri, R., Grosso, G., and Caraci, F. (2021). The Therapeutic Potential of Carnosine\/Anserine Supplementation against Cognitive Decline: A Systematic Review with Meta-Analysis. Biomedicines, 9.","DOI":"10.3390\/biomedicines9030253"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1016\/j.archger.2014.04.008","article-title":"Anserine and carnosine supplementation in the elderly: Effects on cognitive functioning and physical capacity","volume":"59","author":"Szczesniak","year":"2014","journal-title":"Arch. Gerontol. Geriatr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"149","DOI":"10.3233\/JAD-150767","article-title":"Effect of Anserine\/Carnosine Supplementation on Verbal Episodic Memory in Elderly People","volume":"50","author":"Hisatsune","year":"2016","journal-title":"J. Alzheimer\u2019s Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e00541","DOI":"10.1016\/j.neurot.2025.e00541","article-title":"Carnosine supplementation improves cognitive outcomes in younger participants of the NEAT trial","volume":"22","author":"Amraotkar","year":"2025","journal-title":"Neurotherapeutics"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Sch\u00f6n, M., Mousa, A., Berk, M., Chia, W.L., Ukropec, J., Majid, A., Ukropcov\u00e1, B., and de Courten, B. (2019). The Potential of Carnosine in Brain-Related Disorders: A Comprehensive Review of Current Evidence. Nutrients, 11.","DOI":"10.3390\/nu11061196"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4664","DOI":"10.1021\/jf0703809","article-title":"Hydrophilic Chromatographic Determination of Carnosine, Anserine, Balenine, Creatine, and Creatinine","volume":"55","author":"Mora","year":"2007","journal-title":"J. Agric. Food Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1016\/j.chroma.2007.11.075","article-title":"Anserine and carnosine determination in meat samples by pure micellar liquid chromatography","volume":"1189","year":"2008","journal-title":"J. Chromatogr. A"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3275","DOI":"10.3382\/ps.2013-03441","article-title":"Carnosine, anserine, creatine, and inosine 5\u2032-monophosphate contents in breast and thigh meats from 5 lines of Korean native chicken","volume":"92","author":"Jung","year":"2013","journal-title":"Poult. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3555","DOI":"10.1038\/s41598-022-07575-9","article-title":"An attempt to valorize the only black meat chicken breed of India by delineating superior functional attributes of its meat","volume":"12","author":"Sharma","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1817","DOI":"10.5713\/ajas.19.0736","article-title":"Effect of genotypes on macronutrients and antioxidant capacity of chicken breast meat","volume":"33","author":"Lengkidworraphiphat","year":"2020","journal-title":"Asian-Australas. J. Anim. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Han, Y., Gao, B., Zhao, S., Wang, M., Jian, L., Han, L., and Liu, X. (2019). Simultaneous Detection of Carnosine and Anserine by UHPLC-MS\/MS and Its Application on Biomarker Analysis for Differentiation of Meat and Bone Meal. Molecules, 24.","DOI":"10.3390\/molecules24020217"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"121826","DOI":"10.1016\/j.jchromb.2019.121826","article-title":"LC-ESI-MS\/MS quantification of carnosine, anserine, and balenine in meat samples","volume":"1132","author":"Uenoyama","year":"2019","journal-title":"J. Chromatogr. B"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"124660","DOI":"10.1016\/j.jchromb.2025.124660","article-title":"Rapid separation and quantification of imidazole dipeptides in meats using a PBr column packed with 3-(pentabromobenzyloxy)propyl group modified silica gel","volume":"1262","author":"Ozaki","year":"2025","journal-title":"J. Chromatogr. B"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"142445","DOI":"10.1016\/j.foodchem.2024.142445","article-title":"Development and validation of a quantitative method for discriminating Korean and Japanese red seabream (Pagrus major) via UHPLC\u2013QqQ\u2013MS\/MS","volume":"468","author":"Lee","year":"2025","journal-title":"Food Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"36","DOI":"10.17221\/192\/2019-CJFS","article-title":"Monitoring of imidazole dipeptides in meat products by capillary zone electrophoresis","volume":"38","author":"Szerdahelyi","year":"2020","journal-title":"Czech. J. Food Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6548","DOI":"10.1016\/S1452-3981(23)06741-X","article-title":"Voltammetric Characterization and Determination of Histidine Dipeptides\u2014Carnosine and Anserine","volume":"10","year":"2015","journal-title":"Int. J. Electrochem. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.chroma.2004.02.066","article-title":"Determination of carnosine in feed and meat by high-performance anion-exchange chromatography with integrated pulsed amperometric detection","volume":"1035","author":"Nardiello","year":"2004","journal-title":"J. Chromatogr. A"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1006\/fstl.1999.0602","article-title":"HPLC Determination of Carnosine in Commercial Canned Soups and Natural Meat Extracts","volume":"33","author":"Takeuchi","year":"2000","journal-title":"LWT-Food Sci. Technol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/0378-4347(92)80361-S","article-title":"Determination of carnosine and other biogenic imidazoles in equine plasma by isocratic reversed-phase ion-pair high-performance liquid chromatography","volume":"579","author":"Dunnett","year":"1992","journal-title":"J. Chromatogr. B Biomed. Sci. Appl."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"139746","DOI":"10.1016\/j.cplett.2022.139746","article-title":"Theoretical insights into the linear relationship between pKa values and vibrational frequencies","volume":"803","author":"Quintano","year":"2022","journal-title":"Chem. Phys. Lett."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"104323","DOI":"10.1016\/j.jfca.2021.104323","article-title":"Development of a quantitative method for analyzing three imidazole dipeptides using high-performance liquid chromatography and its application for meat and fish","volume":"106","author":"Shinya","year":"2022","journal-title":"J. Food Compos. Anal."},{"key":"ref_37","unstructured":"Latimer, G.W. (2023). AOAC SMPR\u00ae 2025.003: Standard Method Performance Requirements (SMPRs\u00ae) for Determination of Selected Trace Elements in Food and Beverages. Official Methods of Analysis of AOAC INTERNATIONAL, Oxford University Press. [22nd ed.]."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"103590","DOI":"10.1016\/j.psj.2024.103590","article-title":"Genome-wide association studies of anserine and carnosine contents in the breast meat of Korean native chickens","volume":"103","author":"Kim","year":"2024","journal-title":"Poult. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Khumpeerawat, P., Duangjinda, M., and Phasuk, Y. (2021). Carnosine Content and Its Association with Carnosine-Related Gene Expression in Breast Meat of Thai Native and Black-Bone Chicken. Animals, 11.","DOI":"10.3390\/ani11071987"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"101776","DOI":"10.1016\/j.psj.2022.101776","article-title":"Effects of \u03b2-alanine and L-histidine supplementation on carnosine contents in and quality and secondary structure of proteins in slow-growing Korat chicken meat","volume":"101","author":"Suwanvichanee","year":"2022","journal-title":"Poult. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"5680","DOI":"10.1002\/jsfa.9829","article-title":"Differences in muscle histidine-containing dipeptides in broilers","volume":"99","author":"Barbaresi","year":"2019","journal-title":"J. Sci. Food Agric."}],"container-title":["Methods and Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2409-9279\/9\/5\/131\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T04:41:36Z","timestamp":1788669696000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2409-9279\/9\/5\/131"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,4]]},"references-count":41,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,10]]}},"alternative-id":["mps9050131"],"URL":"https:\/\/doi.org\/10.3390\/mps9050131","relation":{},"ISSN":["2409-9279"],"issn-type":[{"value":"2409-9279","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,4]]}}}