{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T15:56:01Z","timestamp":1792598161990,"version":"4.5.0"},"reference-count":50,"publisher":"American Chemical Society (ACS)","issue":"41","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/na","name":"National Basic Research Program of China","doi-asserted-by":"publisher","award":["2025YFA1805602"],"award-info":[{"award-number":["2025YFA1805602"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22225404"],"award-info":[{"award-number":["22225404"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,21]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Fatty acids (FAs), as key lipids in energy storage and signaling, are dynamically regulated to maintain cellular homeostasis. FA profiling has become a valuable approach for investigating lipid reprogramming in disease. Although isomer-resolved mass spectrometry (MS) methods facilitate differentiation of FA isomers, quantitation is limited by the scarcity of isomer standards. Here, leveraging isobaric intramolecular Paterno\u0300\u2013Bu\u0308chi (i2PB) reactions, we develop an isomer-resolved multiplexed quantitation method for FA profiling. The i2PB reagents, synthesized via amidation of benzoyl-phenylalanine with the 6-plex TMT tags at near-quantitative conversion, overcome the low-yield limitations of conventional PB reactions for low-abundance lipids. Rapid i2PB reactions (4 s) enable online integration with liquid chromatography, while subsequent MS2 and MS3 analyses generate reporter ions for 6-channel FA quantitation at the sum composition and C\u2550C location levels, respectively. For FA quantitation, this approach offers improved throughput, amol sensitivity, and a linear dynamic range of 2 orders of magnitude. Using i2PB, we achieve parallel quantitation of over 100 FAs with defined C\u2550C locations in normal and breast cancer cells, providing new insights into metabolic reprogramming of FAs in breast cancer cells.<\/jats:p>","DOI":"10.1021\/jacs.6c13601","type":"journal-article","created":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T10:01:43Z","timestamp":1791367303000},"page":"44629-44638","source":"Crossref","is-referenced-by-count":0,"title":["Isobaric Intramolecular\nPaterno\u0300\u2013Bu\u0308chi\nReactions Enable Multiplexed Quantitation of Fatty Acids down to C\u2550C\nLocations"],"prefix":"10.1021","volume":"148","author":[{"given":"Lipeng","family":"Qiao","sequence":"first","affiliation":[{"name":"Tsinghua University , , ,","place":["Beijing, China, 100084"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Zhao","sequence":"additional","affiliation":[{"name":"Tsinghua University , , ,","place":["Beijing, China, 100084"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8694-9900","authenticated-orcid":true,"given":"Yu","family":"Xia","sequence":"additional","affiliation":[{"name":"Tsinghua University , , ,","place":["Beijing, China, 100084"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,7]]},"reference":[{"key":"2026102111091161000_cit1","doi-asserted-by":"publisher","first-page":"1321","DOI":"10.1038\/s41589-020-00688-0","article-title":"Structural and functional consequences\nof reversible lipid asymmetry in living membranes","volume":"16","author":"Doktorova","year":"2020","journal-title":"Nat. Chem. Biol."},{"key":"2026102111091161000_cit2","doi-asserted-by":"publisher","first-page":"124","DOI":"10.1016\/j.plipres.2013.12.001","article-title":"Regulation of energy\nmetabolism by\nlong-chain fatty acids","volume":"53","author":"Nakamura","year":"2014","journal-title":"Prog. Lipid Res."},{"key":"2026102111091161000_cit3","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1038\/nm.4073","article-title":"Inflammatory signaling\nin human tuberculosis granulomas\nis spatially organized","volume":"22","author":"Marakalala","year":"2016","journal-title":"Nat. Med."},{"key":"2026102111091161000_cit4","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1038\/nrm.2017.138","article-title":"Understanding the diversity\nof membrane lipid composition","volume":"19","author":"Harayama","year":"2018","journal-title":"Nat. Rev. Mol.\nCell Biol."},{"key":"2026102111091161000_cit5","doi-asserted-by":"publisher","first-page":"101242","DOI":"10.1016\/j.plipres.2023.101242","article-title":"FADS2 function at\nthe major cancer hotspot 11q13 locus alters fatty acid metabolism\nin cancer","volume":"92","author":"Kothapalli","year":"2023","journal-title":"Prog. Lipid Res."},{"key":"2026102111091161000_cit6","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1038\/s41586-019-0904-1","article-title":"Evidence for an alternative fatty\nacid desaturation pathway increasing cancer plasticity","volume":"566","author":"Vriens","year":"2019","journal-title":"Nature"},{"key":"2026102111091161000_cit7","doi-asserted-by":"publisher","first-page":"648","DOI":"10.1016\/j.bbalip.2011.07.006","article-title":"High sensitivity\nquantitative lipidomics\nanalysis of fatty acids in biological samples by gas chromatography-mass\nspectrometry","volume":"1811","author":"Quehenberger","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"2026102111091161000_cit8","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1016\/j.jfda.2019.10.003","article-title":"Gas chromatography-mass\nspectrometry-based analytical\nstrategies for fatty acid analysis in biological samples","volume":"28","author":"Chiu","year":"2020","journal-title":"J. Food Drug Anal."},{"key":"2026102111091161000_cit9","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1002\/1438-9312(200201)104:1&lt;36::AID-EJLT111136&gt;3.0.CO;2-W","article-title":"Mass spectrometry of fatty acid derivatives","volume":"104","author":"Dobson","year":"2002","journal-title":"Eur. J. Lipid Sci. Technol."},{"issue":"1","key":"2026102111091161000_cit10","doi-asserted-by":"publisher","first-page":"3940","DOI":"10.1038\/s41467-023-39617-9","article-title":"Ozone-enabled fatty acid discovery reveals\nunexpected diversity in the human lipidome","volume":"14","author":"Menzel","year":"2023","journal-title":"Nat. Commun."},{"key":"2026102111091161000_cit11","doi-asserted-by":"publisher","first-page":"15681","DOI":"10.1021\/jacs.7b06416","article-title":"Pinpointing Double\nBond and sn-Positions in Glycerophospholipids via Hybrid 193 nm Ultraviolet\nPhotodissociation (UVPD) Mass Spectrometry","volume":"139","author":"Williams","year":"2017","journal-title":"J. Am. Chem. Soc."},{"key":"2026102111091161000_cit12","doi-asserted-by":"publisher","first-page":"2015","DOI":"10.1194\/jlr.M070177","article-title":"Structural identification\nof triacylglycerol isomers using electron impact excitation of ions\nfrom organics (EIEIO)","volume":"57","author":"Baba","year":"2016","journal-title":"J. Lipid Res."},{"key":"2026102111091161000_cit13","doi-asserted-by":"publisher","first-page":"7230","DOI":"10.1021\/acs.analchem.8b00322","article-title":"Structural Analysis\nof Phospholipid Using Hydrogen Abstraction Dissociation and Oxygen\nAttachment Dissociation in Tandem Mass Spectrometry","volume":"90","author":"Takahashi","year":"2018","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit14","doi-asserted-by":"publisher","first-page":"1981","DOI":"10.1021\/ac981387f","article-title":"Acetonitrile chemical ionization\ntandem mass spectrometry to locate double bonds in polyunsaturated\nfatty acid methyl esters","volume":"71","author":"Van Pelt","year":"1999","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit15","doi-asserted-by":"publisher","first-page":"2592","DOI":"10.1002\/anie.201310699","article-title":"Pinpointing double\nbonds in lipids by Paterno-Buchi reactions and\nmass spectrometry","volume":"53","author":"Ma","year":"2014","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026102111091161000_cit16","doi-asserted-by":"publisher","first-page":"2573","DOI":"10.1073\/pnas.1523356113","article-title":"Identification and\nquantitation of\nlipid C = C location isomers: A shotgun lipidomics approach enabled\nby photochemical reaction","volume":"113","author":"Ma","year":"2016","journal-title":"Proc. Natl. Acad.\nSci. U. S. A."},{"key":"2026102111091161000_cit17","doi-asserted-by":"publisher","first-page":"11905","DOI":"10.1021\/acs.analchem.9b02667","article-title":"Deep Lipidomics and Molecular Imaging\nof Unsaturated Lipid Isomers: A Universal Strategy Initiated by mCPBA\nEpoxidation","volume":"91","author":"Kuo","year":"2019","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit18","doi-asserted-by":"publisher","first-page":"1791","DOI":"10.1021\/acs.analchem.8b04905","article-title":"Identification of Double\nBond Position Isomers in Unsaturated Lipids by m-CPBA Epoxidation\nand Mass Spectrometry Fragmentation","volume":"91","author":"Feng","year":"2019","journal-title":"Anal. Chem."},{"issue":"39","key":"2026102111091161000_cit19","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202207098","article-title":"Lipid Mass Tags via\nAziridination for Probing Unsaturated Lipid Isomers and Accurate Relative\nQuantification","volume":"61","author":"Yang","year":"2022","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026102111091161000_cit20","doi-asserted-by":"publisher","first-page":"6216","DOI":"10.1021\/acs.analchem.1c05607","article-title":"Chloramine-T-Enabled\nMass Spectrometric Analysis of C = C Isomers of Unsaturated Fatty\nAcids and Phosphatidylcholines in Human Thyroids","volume":"94","author":"Zhang","year":"2022","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit21","doi-asserted-by":"publisher","first-page":"4263","DOI":"10.1038\/s41467-023-40046-x","article-title":"Deep-profiling of phospholipidome\nvia rapid orthogonal separations and isomer-resolved mass spectrometry","volume":"14","author":"Xia","year":"2023","journal-title":"Nat. Commun."},{"key":"2026102111091161000_cit22","doi-asserted-by":"publisher","first-page":"693","DOI":"10.1002\/mas.21492","article-title":"Selection\nof internal standards for\naccurate quantification of complex lipid species in biological extracts\nby electrospray ionization mass spectrometry-What, how and why?","volume":"36","author":"Wang","year":"2017","journal-title":"Mass Spectrom. Rev."},{"key":"2026102111091161000_cit23","doi-asserted-by":"publisher","first-page":"5293","DOI":"10.1021\/pr500880b","article-title":"Isobaric labeling-based relative\nquantification in shotgun proteomics","volume":"13","author":"Rauniyar","year":"2014","journal-title":"J. Proteome\nRes."},{"issue":"8","key":"2026102111091161000_cit24","doi-asserted-by":"publisher","first-page":"1895","DOI":"10.1021\/ac0262560","article-title":"Tandem mass tags: a\nnovel quantification strategy for comparative analysis of complex\nprotein mixtures by MS\/MS","volume":"75","author":"Thompson","year":"2003","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit25","doi-asserted-by":"publisher","first-page":"399","DOI":"10.1038\/s41592-020-0781-4","article-title":"TMTpro reagents:\na set of isobaric labeling mass tags enables simultaneous proteome-wide\nmeasurements across 16 samples","volume":"17","author":"Li","year":"2020","journal-title":"Nat. Methods"},{"key":"2026102111091161000_cit26","doi-asserted-by":"publisher","first-page":"1154","DOI":"10.1074\/mcp.M400129-MCP200","article-title":"Multiplexed protein quantitation\nin Saccharomyces cerevisiae using amine-reactive isobaric tagging\nreagents","volume":"3","author":"Ross","year":"2004","journal-title":"Mol. Cell. Proteomics"},{"key":"2026102111091161000_cit27","doi-asserted-by":"publisher","first-page":"3651","DOI":"10.1002\/pmic.200700316","article-title":"8-plex\nquantitation of changes in cerebrospinal fluid protein expression\nin subjects undergoing intravenous immunoglobulin treatment for Alzheimer\u2019s\ndisease","volume":"7","author":"Choe","year":"2007","journal-title":"Proteomics"},{"key":"2026102111091161000_cit28","doi-asserted-by":"publisher","first-page":"235","DOI":"10.1016\/bs.mie.2021.10.011","article-title":"Multiplexed quantitative neuropeptidomics\nvia DiLeu isobaric tagging","volume-title":"Methods in Enzymology","author":"Sauer","year":"2022"},{"key":"2026102111091161000_cit29","doi-asserted-by":"publisher","first-page":"8228","DOI":"10.1021\/acs.analchem.0c00473","article-title":"21-plex DiLeu Isobaric\nTags for High-Throughput\nQuantitative Proteomics","volume":"92","author":"Frost","year":"2020","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit30","doi-asserted-by":"publisher","first-page":"1606","DOI":"10.1021\/acs.jproteome.7b00896","article-title":"Systematic\nAnalysis of Fatty Acids\nin Human Cells with a Multiplexed Isobaric Tag (TMT)-Based Method","volume":"17","author":"Sun","year":"2018","journal-title":"J. Proteome Res."},{"key":"2026102111091161000_cit31","doi-asserted-by":"publisher","first-page":"762","DOI":"10.1038\/s41557-023-01436-2","article-title":"Diazobutanone-assisted\nisobaric labelling\nof phospholipids and sulfated glycolipids enables multiplexed quantitative\nlipidomics using tandem mass spectrometry","volume":"16","author":"Gu","year":"2024","journal-title":"Nat.\nChem."},{"key":"2026102111091161000_cit32","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1039\/D2AN01699K","article-title":"Isobaric 6-plex and tosyl dual tagging for the determination\nof positional isomers and quantitation of monounsaturated fatty acids\nusing rapid UHPLC-MS\/MS","volume":"148","author":"Armbruster","year":"2023","journal-title":"Analyst"},{"key":"2026102111091161000_cit33","doi-asserted-by":"publisher","first-page":"13036","DOI":"10.1021\/acs.analchem.2c01917","article-title":"Quantitative Analysis\nand Structural Elucidation of Fatty Acids by Isobaric Multiplex Labeling\nReagents for Carbonyl-Containing Compound (SUGAR) Tags and m-CPBA\nEpoxidation","volume":"94","author":"Feng","year":"2022","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit34","doi-asserted-by":"publisher","first-page":"5117","DOI":"10.1021\/acs.analchem.3c00085","article-title":"Visible-Light\nPaterno-Buchi Reaction for Lipidomic Profiling at Detailed Structure\nLevels","volume":"95","author":"Shi","year":"2023","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit35","doi-asserted-by":"publisher","first-page":"1523","DOI":"10.1194\/jlr.M033506","article-title":"Shorthand\nnotation for lipid structures derived from mass spectrometry","volume":"54","author":"Liebisch","year":"2013","journal-title":"J. Lipid Res."},{"key":"2026102111091161000_cit36","doi-asserted-by":"publisher","first-page":"13470","DOI":"10.1021\/acs.analchem.0c02896","article-title":"Next-Generation\nPaterno-Buchi Reagents for Lipid Analysis by Mass Spectrometry","volume":"92","author":"Zhao","year":"2020","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit37","doi-asserted-by":"publisher","first-page":"7173","DOI":"10.1021\/acs.analchem.9b00374","article-title":"Analysis of Conjugated Fatty Acid Isomers by the Paterno-Buchi\nReaction\nand Trapped Ion Mobility Mass Spectrometry","volume":"91","author":"Xie","year":"2019","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit38","doi-asserted-by":"publisher","first-page":"555","DOI":"10.1021\/acs.analchem.4c04797","article-title":"Tracking the\nGeometric and Positional Isomerization of Lipid C horizontal lineC\nBonds in the Bacterial Stress Responses by Mass Spectrometry","volume":"97","author":"Fu","year":"2025","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit39","doi-asserted-by":"publisher","first-page":"3544","DOI":"10.1021\/acs.analchem.3c05431","article-title":"Studies of Labware Contamination\nduring Lipid Extraction in Mass\nSpectrometry-Based Lipidome Analysis","volume":"96","author":"Canez","year":"2024","journal-title":"Anal.\nChem."},{"key":"2026102111091161000_cit40","doi-asserted-by":"publisher","first-page":"100110","DOI":"10.1016\/j.jlr.2021.100110","article-title":"A liquid chromatography-mass spectrometry\nworkflow for in-depth quantitation of fatty acid double bond location\nisomers","volume":"62","author":"Zhao","year":"2021","journal-title":"J. Lipid Res."},{"key":"2026102111091161000_cit41","doi-asserted-by":"publisher","first-page":"7150","DOI":"10.1021\/ac502040v","article-title":"MultiNotch\nMS3 enables accurate, sensitive, and multiplexed detection of differential\nexpression across cancer cell line proteomes","volume":"86","author":"McAlister","year":"2014","journal-title":"Anal. Chem."},{"key":"2026102111091161000_cit42","doi-asserted-by":"publisher","first-page":"763","DOI":"10.1038\/nrc2222","article-title":"Fatty acid\nsynthase and the lipogenic phenotype in cancer pathogenesis","volume":"7","author":"Menendez","year":"2007","journal-title":"Nat. Rev. Cancer"},{"key":"2026102111091161000_cit43","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1038\/s41585-022-00654-6","article-title":"Fatty acid\nmetabolism reprogramming in ccRCC: mechanisms and potential targets","volume":"20","author":"Tan","year":"2023","journal-title":"Nat. Rev. Urol."},{"key":"2026102111091161000_cit44","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1038\/s41568-022-00537-3","article-title":"STAT proteins\nin cancer: orchestration of metabolism","volume":"23","author":"Li","year":"2023","journal-title":"Nat.\nRev. Cancer"},{"key":"2026102111091161000_cit45","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.plefa.2018.05.002","article-title":"A rare eicosanoid\nprecursor analogue, sciadonic acid (5Z,11Z,14Z-20:3), detected in\nvivo in hormone positive breast cancer tissue","volume":"134","author":"Park","year":"2018","journal-title":"Prostaglandins, Leukotrienes Essent. Fatty Acids"},{"key":"2026102111091161000_cit46","doi-asserted-by":"publisher","first-page":"100410","DOI":"10.1016\/j.jlr.2023.100410","article-title":"Alternative\nfatty acid desaturation pathways revealed by deep profiling\nof total fatty acids in RAW 264.7 cell line","volume":"64","author":"Xia","year":"2023","journal-title":"J. Lipid Res."},{"key":"2026102111091161000_cit47","doi-asserted-by":"publisher","first-page":"537","DOI":"10.1007\/s11745-010-3436-x","article-title":"Characteristics of the fatty acid composition of a\ndeep-sea vent gastropod, Ifremeria nautilei","volume":"45","author":"Saito","year":"2010","journal-title":"Lipids"},{"key":"2026102111091161000_cit48","doi-asserted-by":"publisher","first-page":"bqz046","DOI":"10.1210\/endocr\/bqz046","article-title":"CPT1A-mediated Fat Oxidation, Mechanisms, and Therapeutic Potential","volume":"161","author":"Schlaepfer","year":"2020","journal-title":"Endocrinology"},{"key":"2026102111091161000_cit49","doi-asserted-by":"publisher","first-page":"732","DOI":"10.1038\/nrc.2016.89","article-title":"The multifaceted roles\nof fatty acid synthesis in cancer","volume":"16","author":"Ro\u0308hrig","year":"2016","journal-title":"Nat. Rev.\nCancer"},{"key":"2026102111091161000_cit50","doi-asserted-by":"publisher","first-page":"2850","DOI":"10.1158\/0008-5472.CAN-13-0382-T","article-title":"An Essential\nRequirement for the SCAP\/SREBP Signaling Axis to Protect Cancer Cells\nfrom Lipotoxicity","volume":"73","author":"Williams","year":"2013","journal-title":"Cancer Res."}],"container-title":["Journal of the American\nChemical Society"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jacs.6c13601","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/41\/44629\/68767499\/jacs.6c13601.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/41\/44629\/68767499\/jacs.6c13601.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T15:27:53Z","timestamp":1792596473000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jacsat\/article\/148\/41\/44629\/5445870\/Isobaric-Intramolecular-Paterno-Bu-chi-Reactions"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,7]]},"references-count":50,"journal-issue":{"issue":"41","published-online":{"date-parts":[[2026,10,7]]},"published-print":{"date-parts":[[2026,10,21]]}},"URL":"https:\/\/doi.org\/10.1021\/jacs.6c13601","relation":{},"ISSN":["0002-7863","1520-5126"],"issn-type":[{"value":"0002-7863","type":"print"},{"value":"1520-5126","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,7]]}}}