{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T18:29:19Z","timestamp":1791570559310,"version":"4.3.4"},"reference-count":50,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,2,3]],"date-time":"2025-02-03T00:00:00Z","timestamp":1738540800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Japan Racing Association"},{"name":"Ito Foundation"},{"name":"Agriculture and Livestock Industries Co., Ltd."}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Background: Japanese Black beef is known for its high intramuscular fat content, an important factor in its distinctive Wagyu aroma. Wet aging, which involves vacuum-packing meat and storing it at low temperatures, enhances flavor, texture, and tenderness and is essential for maintaining and improving meat quality. In this study, changes in metabolites and lipid profiles were investigated during the wet aging of Japanese Black and Holstein beef. Methods\/Results: Gas chromatography\u2013mass spectrometry identified 113 metabolites in Japanese Black beef and 94 in Holstein beef, with significant increases in metabolites like aspartic acid and maleic acid over the aging period. Regarding lipid composition, total free fatty acids significantly increased with wet aging, with Japanese Black beef showing significantly higher concentrations of oleic and linoleic acids than Holstein beef. Additionally, lipid analysis by liquid chromatography\u2013mass spectrometry revealed a reduction in specific phospholipids, particularly lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), with notable decreases in LPC (18:1), LPC (18:2), LPE (18:1), and LPE (18:2). Conclusions: These results suggest that wet aging influences the stability of membrane lipids, facilitating the degradation of phospholipids into free fatty acids, and improving the flavor of Japanese Black beef.<\/jats:p>","DOI":"10.3390\/metabo15020094","type":"journal-article","created":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T05:13:43Z","timestamp":1738646023000},"page":"94","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Metabolites and Free Fatty Acids in Japanese Black Beef During Wet Aging"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5702-4471","authenticated-orcid":false,"given":"Shuji","family":"Ueda","sequence":"first","affiliation":[{"name":"Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University, Kobe 657-8501, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuka","family":"Yoshida","sequence":"additional","affiliation":[{"name":"Japan Meat Science and Technology Institute, Inc., Tokyo 150-0013, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuka","family":"Tateoka","sequence":"additional","affiliation":[{"name":"Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University, Kobe 657-8501, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5774-9539","authenticated-orcid":false,"given":"Biniam","family":"Kebede","sequence":"additional","affiliation":[{"name":"Department of Food Science, University of Guelph, Guelph, ON N1G 2W1, Canada"},{"name":"Department of Food Science, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masakazu","family":"Shinohara","sequence":"additional","affiliation":[{"name":"The Integrated Center for Mass Spectrometry, Kobe University, Kobe 650-0017, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroki","family":"Nakanishi","sequence":"additional","affiliation":[{"name":"Lipidome Lab Co., Ltd., Akita 010-0825, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5969-4295","authenticated-orcid":false,"given":"Itsuko","family":"Fukuda","sequence":"additional","affiliation":[{"name":"Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University, Kobe 657-8501, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0654-5461","authenticated-orcid":false,"given":"Yasuhito","family":"Shirai","sequence":"additional","affiliation":[{"name":"Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University, Kobe 657-8501, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"933","DOI":"10.5713\/ajas.18.0333","article-title":"The Japanese Wagyu beef industry: Current situation and future prospects\u2014A review","volume":"31","author":"Gotoh","year":"2018","journal-title":"Asian-Australas J. Anim. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1080\/1828051X.2023.2216712","article-title":"Biodiversity and genetics of beef quality, a review","volume":"22","author":"Bittante","year":"2023","journal-title":"Ital. J. Anim. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e13841","DOI":"10.1111\/asj.13841","article-title":"Effect of intramuscular fat content on the sensory characteristics and dynamic flavor attributes of Japanese black cattle beef","volume":"94","author":"Hirai","year":"2023","journal-title":"Anim. Sci. J."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ueda, S., Yamanoue, M., Sirai, Y., and Iwamoto, E. (2021). Exploring the Characteristic Aroma of Beef from Japanese Black Cattle (Japanese Wagyu) via Sensory Evaluation and Gas Chromatography-Olfactometry. Metabolites, 11.","DOI":"10.3390\/metabo11010056"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1292\/jvms.21-0441","article-title":"Congenital syringohydromyelia in a crossbred (Holstein-Friesian \u00d7 Japanese Black) beef calf","volume":"84","author":"Ishiyama","year":"2022","journal-title":"J. Vet. Med. Sci.\/Jpn. Soc. Vet. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e70073","DOI":"10.1111\/1541-4337.70073","article-title":"Superheated steam technology: Recent developments and applications in food industries","volume":"23","author":"Zhang","year":"2024","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Huang, W., Guo, Y., Du, W., Zhang, X., Li, A., and Miao, X. (2017). Global transcriptome analysis identifies differentially expressed genes related to lipid metabolism in Wagyu and Holstein cattle. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-05702-5"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Yang, Z., Ma, X., Zhang, D., Li, B., Gao, N., Li, X., Mei, C., and Zan, L. (2024). Bta-miR-330 promotes bovine intramuscular pre-adipocytes adipogenesis via targeting SESN3 to activate the Akt-mTOR signaling pathway. Int. J. Biol. Macromol., 275.","DOI":"10.1016\/j.ijbiomac.2024.133650"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Yamada, T., Kamiya, M., and Higuchi, M. (2020). Gas Chromatography-Mass Spectrometry-Based Metabolomic Analysis of Wagyu and Holstein Beef. Metabolites, 10.","DOI":"10.3390\/metabo10030095"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1080\/09168451.2018.1528139","article-title":"Comparative metabolomics of Japanese Black cattle beef and other meats using gas chromatography\u2013mass spectrometry","volume":"83","author":"Ueda","year":"2019","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.5187\/jast.2023.e110","article-title":"Aging mechanism for improving the tenderness and taste characteristics of meat","volume":"65","author":"Joo","year":"2023","journal-title":"J. Anim. Sci. Technol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"953","DOI":"10.5851\/kosfa.2022.e63","article-title":"Effects of Aging Methods and Periods on Quality Characteristics of Beef","volume":"42","author":"Kim","year":"2022","journal-title":"Food Sci. Anim. Resour."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e17091","DOI":"10.1016\/j.heliyon.2023.e17091","article-title":"Analysis of physicochemical properties of dry-cured beef made from Hanwoo and Holstein meat distributed in South Korea","volume":"9","author":"Lee","year":"2023","journal-title":"Heliyon"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1332","DOI":"10.5713\/ajas.19.0589","article-title":"Meat quality characteristics of the Arabian camel (Camelus dromedarius) at different ages and post-mortem ageing periods","volume":"33","author":"Suliman","year":"2020","journal-title":"Asian-Australas J. Anim. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"589","DOI":"10.5851\/kosfa.2021.e27","article-title":"Endogenous Proteolytic Systems and Meat Tenderness: Influence of Post-Mortem Storage and Processing","volume":"41","author":"Kaur","year":"2021","journal-title":"Food Sci. Anim. Resour."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"374","DOI":"10.5713\/ab.22.0370","article-title":"Invited Review\u2014Postmortem skeletal muscle metabolism of farm animals approached with metabolomics","volume":"36","author":"Muroya","year":"2023","journal-title":"Anim. Biosci."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Di Paolo, M., Ambrosio, R.L., Lambiase, C., Vuoso, V., Salzano, A., Bifulco, G., Barone, C.M.A., and Marrone, R. (2023). Effects of the Aging Period and Method on the Physicochemical, Microbiological and Rheological Characteristics of Two Cuts of Charolais Beef. Foods, 12.","DOI":"10.3390\/foods12030531"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Chai, W., Wang, L., Li, T., Wang, T., Wang, X., Yan, M., Zhu, M., Gao, J., Wang, C., and Ma, Q. (2024). Liquid Chromatography-Mass Spectrometry-Based Metabolomics Reveals Dynamic Metabolite Changes during Early Postmortem Aging of Donkey Meat. Foods, 13.","DOI":"10.3390\/foods13101466"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Ueda, S., Yoshida, Y., Kebede, B., Kitamura, C., Sasaki, R., Shinohara, M., Fukuda, I., and Shirai, Y. (2024). New Implications of Metabolites and Free Fatty Acids in Quality Control of Crossbred Wagyu Beef during Wet Aging Cold Storage. Metabolites, 14.","DOI":"10.3390\/metabo14020095"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Shahidi, F., and Hossain, A. (2022). Role of Lipids in Food Flavor Generation. Molecules, 27.","DOI":"10.3390\/molecules27155014"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"17536","DOI":"10.1021\/acs.jafc.4c05295","article-title":"Lipid Hydrolysis, Oxidation, and Fatty Acid Formation Pathway Mapping of Synergistically Fermented Sausage and Characterization of Lipid Mediating Genes","volume":"72","author":"Woldemariam","year":"2024","journal-title":"J. Agric. Food Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/S0021-9258(18)64849-5","article-title":"A simple method for the isolation and purification of total lipides from animal tissues","volume":"226","author":"Folch","year":"1957","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1007\/978-1-60761-322-0_15","article-title":"Qualitative and Quantitative Analyses of Phospholipids by LC\u2013MS for Lipidomics","volume":"579","author":"Nakanishi","year":"2009","journal-title":"Methods Mol. Biol. (Clifton N.J.)"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Muroya, S., Horiuchi, Y., Iguchi, K., Higuchi, T., Sakamoto, S., Ojima, K., and Matsukawa, K. (2024). Depth of Interbreed Difference in Postmortem Bovine Muscle Determined by CE-FT\/MS and LC-FT\/MS Metabolomics. Metabolites, 14.","DOI":"10.3390\/metabo14050261"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Sakai, K., Okada, M., and Yamaguchi, S. (2024). Umami and saltiness enhancements of vegetable soup by enzyme-produced glutamic acid and branched-chain amino acids. Front. Nutr., 11.","DOI":"10.3389\/fnut.2024.1436113"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/s41538-023-00178-2","article-title":"The flavor-enhancing action of glutamate and its mechanism involving the notion of kokumi","volume":"7","author":"Yamamoto","year":"2023","journal-title":"NPJ Sci. Food"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1111\/j.1740-0929.2009.00648.x","article-title":"Effects of the fattening period on the fatty acid composition of fat deposits and free amino acid and inosinic acid contents of the longissimus muscle in carcasses of Japanese Black steers","volume":"80","author":"Iwamoto","year":"2009","journal-title":"Anim. Sci. J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1111\/1750-3841.16101","article-title":"Molecular insights into human taste perception and umami tastants: A review","volume":"87","author":"Diepeveen","year":"2022","journal-title":"J. Food Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"109621","DOI":"10.1016\/j.meatsci.2024.109621","article-title":"Lipidome and metabolome profiling of longissimus lumborum beef with different ultimate pH and postmortem aging","volume":"217","author":"Ribeiro","year":"2024","journal-title":"Meat Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"54","DOI":"10.5851\/kosfa.2019.e3","article-title":"Effects of Aging and Aging Method on Physicochemical and Sensory Traits of Different Beef Cuts","volume":"39","author":"Kim","year":"2019","journal-title":"Food Sci. Anim. Resour."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2129","DOI":"10.1080\/10408398.2021.1971156","article-title":"The effect of aging on beef taste, aroma and texture, and the role of microorganisms: A review","volume":"63","author":"Xu","year":"2023","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Dom\u00ednguez, R., Pateiro, M., Gagaoua, M., Barba, F.J., Zhang, W., and Lorenzo, J.M. (2019). A Comprehensive Review on Lipid Oxidation in Meat and Meat Products. Antioxidants, 8.","DOI":"10.3390\/antiox8100429"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Ueda, S., Hosoda, M., Kasamatsu, K., Horiuchi, M., Nakabayashi, R., Kang, B., Shinohara, M., Nakanishi, H., Ohto-Nakanishi, T., and Yamanoue, M. (2022). Production of Hydroxy Fatty Acids, Precursors of \u03b3-Hexalactone, Contributes to the Characteristic Sweet Aroma of Beef. Metabolites, 12.","DOI":"10.3390\/metabo12040332"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1152\/physrev.00061.2021","article-title":"A systematic review of the biological mediators of fat taste and smell","volume":"103","author":"Brooks","year":"2023","journal-title":"Physiol. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1242\/jeb.124941","article-title":"The sarcomeric cytoskeleton: From molecules to motion","volume":"219","author":"Gautel","year":"2016","journal-title":"J. Exp. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1111\/j.1365-2621.2003.tb14121.x","article-title":"Relationship Between Weakening of Z-disks and Liberation of Phospholipids During Postmortem Aging of Pork and Beef","volume":"68","author":"Ahn","year":"2003","journal-title":"J. Food Sci."},{"key":"ref_37","first-page":"221","article-title":"Adipose Triglyceride Lipase Regulation: An Overview","volume":"19","author":"Cerk","year":"2018","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Ueda, S., Hosoda, M., Yoshino, K.-i., Yamanoue, M., and Shirai, Y. (2021). Gene Expression Analysis Provides New Insights into the Mechanism of Intramuscular Fat Formation in Japanese Black Cattle. Genes, 12.","DOI":"10.3390\/genes12081107"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1016\/j.jbior.2017.10.009","article-title":"Review of four major distinct types of human phospholipase A(2)","volume":"67","author":"Vasquez","year":"2018","journal-title":"Adv. Biol. Regul."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"109402","DOI":"10.1016\/j.meatsci.2023.109402","article-title":"Genome-wide scans identify biological and metabolic pathways regulating carcass and meat quality traits in beef cattle","volume":"209","author":"Arikawa","year":"2024","journal-title":"Meat Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"587","DOI":"10.3892\/ijmm.2014.2047","article-title":"Phylogenetic and structural analysis of the phospholipase A2 gene family in vertebrates","volume":"35","author":"Huang","year":"2015","journal-title":"Int. J. Mol. Med."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Khan, S.A., and Ilies, M.A. (2023). The Phospholipase A2 Superfamily: Structure, Isozymes, Catalysis, Physiologic and Pathologic Roles. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24021353"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"22030","DOI":"10.1074\/jbc.272.35.22030","article-title":"Regiospecificity and catalytic triad of lysophospholipase I","volume":"272","author":"Wang","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"131139","DOI":"10.1016\/j.foodchem.2021.131139","article-title":"Dissecting grilled red and white meat flavor: Its characteristics, production mechanisms, influencing factors and chemical hazards","volume":"371","author":"Bassam","year":"2022","journal-title":"Food Chem."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Oh, D.K., Lee, T.E., Lee, J., Shin, K.C., and Park, J.B. (2024). Biocatalytic oxyfunctionalization of unsaturated fatty acids to oxygenated chemicals via hydroxy fatty acids. Biotechnol. Adv., 79.","DOI":"10.1016\/j.biotechadv.2024.108510"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"15325","DOI":"10.1021\/acs.jafc.2c04527","article-title":"Saturated Linear Aliphatic \u03b3- and \u03b4-Lactones in Wine: A Review","volume":"70","author":"Miller","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"20738","DOI":"10.1021\/acs.jafc.4c05591","article-title":"Mechanochemistry in Glycation Research","volume":"72","author":"Xing","year":"2024","journal-title":"J. Agric. Food Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e13813","DOI":"10.1111\/asj.13813","article-title":"Comparison of 2,5-dimethyl-4-hydroxy-3(2H)-furanone, a volatile odor compound generated by the Maillard reaction, in cooked meat of various animal species and parts","volume":"94","author":"Yokoyama","year":"2023","journal-title":"Anim. Sci. J."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1800370","DOI":"10.1002\/ejlt.201800370","article-title":"Decrease in Intramuscular Levels of Phosphatidylethanolamine Bearing Arachidonic Acid During Postmortem Aging Depends on Meat Cuts and Breed","volume":"121","author":"Yamashita","year":"2019","journal-title":"Eur. J. Lipid Sci. Technol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"W147","DOI":"10.1093\/nar\/gkw419","article-title":"Heatmapper: Web-enabled heat mapping for all","volume":"44","author":"Babicki","year":"2016","journal-title":"Nucleic Acids Res."}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/2\/94\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:26:22Z","timestamp":1760027182000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/2\/94"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,3]]},"references-count":50,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["metabo15020094"],"URL":"https:\/\/doi.org\/10.3390\/metabo15020094","relation":{},"ISSN":["2218-1989"],"issn-type":[{"value":"2218-1989","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,3]]}}}