{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T19:57:58Z","timestamp":1790798278451,"version":"4.1.0"},"reference-count":36,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R35GM153450"],"award-info":[{"award-number":["R35GM153450"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000171","name":"Division of Human Resource Development","doi-asserted-by":"publisher","award":["HRD-1547798"],"award-info":[{"award-number":["HRD-1547798"]}],"id":[{"id":"10.13039\/100000171","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000171","name":"Division of Human Resource Development","doi-asserted-by":"publisher","award":["HRD-2111661"],"award-info":[{"award-number":["HRD-2111661"]}],"id":[{"id":"10.13039\/100000171","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,10,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The escalating prevalence and diversity of fentanyl analogues poses an immediate concern for the global community. Fentanyl and its analogues are the primary contributors to both fatal and nonfatal overdoses in the United States. The most recent instances of fentanyl-related overdoses have been attributed to the illicit production of fentanyl, characterized by its exceptionally potent nature. In this study, we present a high-throughput mass spectrometry based method for effective screening of fentanyl analogues with focus on the isomeric separation using commercially available platforms combining liquid chromatography, trapped ion mobility spectrometry, and tandem mass spectrometry (LC-TIMS-q-TOF MS\/MS). The proposed analysis allows for effective separation and identification of 250 synthetic opioids based on the isotopic pattern, retention time, mobility profile, and MS\/MS pattern. Our approach capitalizes on the advancements incorporating parallel accumulation in the mobility trap followed by sequential fragmentation (PASEF) using collision-induced dissociation on the liquid chromatography time scale. While a single chromatography band is commonly observed for single isomeric analogues, a dual mobility band profile attributed to two protonation sites is commonly observed for most fentanyl analogues. Reference mobility values are reported from single standards with 0.2% RSD collected at high resolution (RIMS \u2248 80\u2013120). The added mobility separation resulted in the separation of isomeric compounds without compromising the sensitivity of the LC-q-TOF MS\/MS analysis; that is, a good linear dynamic and (R2 &amp;gt; 0.98) and low limits of detection (LOD) in the 0.08\u20134 ng\/mL range were observed for all synthetic analogues (\u223c100 analogues can be observed with LOD &amp;lt; 1 ng\/mL).<\/jats:p>","DOI":"10.1021\/jasms.5c00203","type":"journal-article","created":{"date-parts":[[2025,9,10]],"date-time":"2025-09-10T04:53:52Z","timestamp":1757480032000},"page":"2206-2212","source":"Crossref","is-referenced-by-count":1,"title":["Complementary Separation\nof Novel Synthetic Opioids"],"prefix":"10.1021","volume":"36","author":[{"given":"Andrew R.","family":"Forero","sequence":"first","affiliation":[{"name":"Florida International University , , , ,","place":["Miami, Florida, United States, 33199"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel A.","family":"Miller","sequence":"additional","affiliation":[{"name":"Florida International University , , , ,","place":["Miami, Florida, United States, 33199"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lilian Valadares","family":"Tose","sequence":"additional","affiliation":[{"name":"Florida International University , , , ,","place":["Miami, Florida, United States, 33199"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew","family":"Willetts","sequence":"additional","affiliation":[{"name":"Bruker Daltonics, Inc. , , ,","place":["Billerica, Massachusetts, United States, 01821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark E.","family":"Ridgeway","sequence":"additional","affiliation":[{"name":"Bruker Daltonics, Inc. , , ,","place":["Billerica, Massachusetts, United States, 01821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-1448-2896","authenticated-orcid":true,"given":"Melvin A.","family":"Park","sequence":"additional","affiliation":[{"name":"Bruker Daltonics, Inc. , , ,","place":["Billerica, Massachusetts, United States, 01821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisa N.","family":"Shoff","sequence":"additional","affiliation":[{"name":"Miami-Dade County Medical Examiner Department , , ,","place":["Miami, Florida, United States, 33330"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1283-4390","authenticated-orcid":true,"given":"Francisco","family":"Fernandez-Lima","sequence":"additional","affiliation":[{"name":"Florida International University , , , ,","place":["Miami, Florida, United States, 33199"]},{"name":"Florida International University , , , ,","place":["Miami, Florida, United States, 33199"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2025,9,10]]},"reference":[{"issue":"5","key":"2026093015110114200_cit1","doi-asserted-by":"publisher","first-page":"1643","DOI":"10.1111\/1556-4029.15282","article-title":"Review of analytical methods for screening and quantification\nof fentanyl analogs and novel synthetic opioids in biological specimens","volume":"68","author":"Palmquist","year":"2023","journal-title":"J. Forensic Sci."},{"key":"2026093015110114200_cit2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.trac.2018.01.007","article-title":"Giulio Mannocchi, Enrico\nMarinelli,\nFrancesco Paolo Busardo, Simona Pichini. New synthetic opioids in\nbiological and non-biological matrices: A review of current analytical\nmethods","volume":"102","author":"Marchei","year":"2018","journal-title":"Trends Anal. Chem."},{"key":"2026093015110114200_cit3","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/j.neubiorev.2018.12.005","article-title":"Fentanyl: Receptor\npharmacology, abuse potential, and\nimplications for treatment","volume":"106","author":"Comer","year":"2019","journal-title":"Neurosci. Biobehav.\nR"},{"issue":"5","key":"2026093015110114200_cit4","doi-asserted-by":"publisher","first-page":"801","DOI":"10.1021\/jm00299a003","article-title":"Structure-Activity\nCorrelations of Morphine-Like Analgetics Based on Efficiencies Following\nIntravenous and Intraventricular Application","volume":"13","author":"Kutter","year":"1970","journal-title":"J. Med. Chem."},{"issue":"4\u20135","key":"2026093015110114200_cit5","first-page":"386","article-title":"Correlations of Structure-Activity\nof Morphine-Like Analgesics Using Regression-Analysis","volume":"266","author":"Kutter","year":"1970","journal-title":"N-S Arch. Pharmakol."},{"issue":"12","key":"2026093015110114200_cit6","doi-asserted-by":"publisher","first-page":"1215","DOI":"10.1016\/j.jpain.2014.08.010","article-title":"The Fentanyl\nStory","volume":"15","author":"Stanley","year":"2014","journal-title":"J. Pain"},{"issue":"33","key":"2026093015110114200_cit7","doi-asserted-by":"publisher","first-page":"837","DOI":"10.15585\/mmwr.mm6533a2","article-title":"Fentanyl Law\nEnforcement Submissions\nand Increases in Synthetic Opioid-Involved Overdose Deaths \u2500\n27 States, 2013\u20132014","volume":"65","author":"Gladden","year":"2016","journal-title":"Morbid. Mortal.\nWeekly Rep."},{"issue":"50 & 51","key":"2026093015110114200_cit8","doi-asserted-by":"publisher","first-page":"1445","DOI":"10.15585\/mmwr.mm655051e1","article-title":"Increases in Drug and\nOpioid-Involved Overdose Deaths \u2500 United States, 2010\u20132015","volume":"65","author":"Rudd","year":"2016","journal-title":"Morbid. Mortal. Weekly Rep."},{"issue":"4","key":"2026093015110114200_cit9","doi-asserted-by":"publisher","first-page":"385","DOI":"10.4155\/fmc.13.215","article-title":"Fentanyl-related\ncompounds and derivatives: current\nstatus and future prospects for pharmaceutical applications","volume":"6","author":"Vardanyan","year":"2014","journal-title":"Future Med. Chem."},{"key":"2026093015110114200_cit10","unstructured":"Krotulski, A. J.; Walton, S. E.; Mohr, A. L. A.; Logan, B. K.\n          2022 Q4 NPS Opioids\nin the United States; The Center for Forensic\nScience Research & Education, 2022."},{"key":"2026093015110114200_cit11","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1021\/jasms.4c00049","article-title":"Identification of\nUnique Fragmentation Patterns of Fentanyl Analog Protomers Using Structures\nfor Lossless Ion Manipulations Ion Mobility-Orbitrap Mass Spectrometry","volume":"35","author":"Hollerbach","year":"2024","journal-title":"J. Am. Soc. Mass Spectrom."},{"issue":"8","key":"2026093015110114200_cit12","doi-asserted-by":"publisher","first-page":"1938","DOI":"10.1080\/10408347.2021.1927668","article-title":"Gas Chromatography-Mass\nSpectrometry Analysis of Synthetic Opioids Belonging to the Fentanyl\nClass: A Review","volume":"52","author":"Valdez","year":"2022","journal-title":"Crit. Rev. Anal. Chem."},{"issue":"6","key":"2026093015110114200_cit13","doi-asserted-by":"publisher","first-page":"490","DOI":"10.1093\/jat\/bkv043","article-title":"An Acute Acetyl Fentanyl Fatality: A Case Report With\nPostmortem Concentrations","volume":"39","author":"McIntyre","year":"2015","journal-title":"J. Anal. Toxicol."},{"key":"2026093015110114200_cit14","unstructured":"Liu, J.; Franckowski, R. E. (Cayman Chemical). Supplemental Material: Differentiation of Isobaric\nand Isomeric Fentanyl Analogs by Gas Chromatography-Mass Spectrometry\n(GC-MS). Poster presented at  AAFS 71st Annual\nScientific Meeting, Baltimore, MD, February 18\u201323, 2019."},{"issue":"12","key":"2026093015110114200_cit15","doi-asserted-by":"publisher","first-page":"2852","DOI":"10.1021\/jasms.1c00267","article-title":"Use of Diagnostic\nIons for the Detection of Fentanyl\nAnalogs in Human Matrices by LC-QTOF","volume":"32","author":"Swanson","year":"2021","journal-title":"J. Am.\nSoc. Mass Spectrom."},{"key":"2026093015110114200_cit16","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jmsacl.2022.07.005","article-title":"Development\nand\napplication of a High-Resolution mass spectrometry method for the\ndetection of fentanyl analogs in urine and serum","volume":"26","author":"Zhang","year":"2022","journal-title":"J. Mass Spectrom Adv. Clin Lab"},{"issue":"21","key":"2026093015110114200_cit17","doi-asserted-by":"publisher","first-page":"6391","DOI":"10.1039\/C9AN01771B","article-title":"Discriminative\npotential of ion mobility spectrometry for the detection of fentanyl\nand fentanyl analogues relative to confounding environmental interferents","volume":"144","author":"Forbes","year":"2019","journal-title":"Analyst"},{"issue":"5","key":"2026093015110114200_cit18","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1007\/s13361-019-02175-w","article-title":"Effects of Solvent Vapor Modifiers\nfor the Separation of Opioid Isomers in Micromachined FAIMS-MS","volume":"30","author":"Wei","year":"2019","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"2026093015110114200_cit19","doi-asserted-by":"publisher","first-page":"338257","DOI":"10.1016\/j.aca.2021.338257","article-title":"Ion mobility-high\nresolution mass spectrometry in anti-doping analysis. Part I: Implementation\nof a screening method with the assessment of a library of substances\nprohibited in sports","volume":"1152","author":"Plachka\u0301","year":"2021","journal-title":"Anal. Chim. Acta"},{"issue":"10","key":"2026093015110114200_cit20","doi-asserted-by":"publisher","first-page":"1904","DOI":"10.1021\/jasms.2c00186","article-title":"A High-Throughput\nIon Mobility Spectrometry-Mass Spectrometry\nScreening Method for Opioid Profiling","volume":"33","author":"Butler","year":"2022","journal-title":"J. Am.\nSoc. Mass Spectrom."},{"issue":"7","key":"2026093015110114200_cit21","doi-asserted-by":"publisher","first-page":"749","DOI":"10.1039\/C7AY02808C","article-title":"Ion mobility\nderived collision cross\nsection as an additional measure to support the rapid analysis of\nabused drugs and toxic compounds using electrospray ion mobility time-of-flight\nmass spectrometry","volume":"10","author":"Lian","year":"2018","journal-title":"Analytical Methods"},{"issue":"1","key":"2026093015110114200_cit22","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s12127-019-00244-0","article-title":"Analyzing\nfentanyl and fentanyl analogues\nby ion mobility spectrometry","volume":"22","author":"Zaknoun","year":"2019","journal-title":"Int. J. Ion Mobil.\nSpectrom."},{"key":"2026093015110114200_cit23","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1016\/j.talanta.2018.02.077","article-title":"Analysis of isomeric opioids in urine\nusing LC-TIMS-TOF MS","volume":"183","author":"Adams","year":"2018","journal-title":"Talanta"},{"key":"2026093015110114200_cit24","doi-asserted-by":"publisher","first-page":"127191","DOI":"10.1016\/j.talanta.2024.127191","article-title":"High-throughput\nscreening of fentanyl analogs","volume":"283","author":"Miller","year":"2025","journal-title":"Talanta"},{"issue":"12","key":"2026093015110114200_cit25","doi-asserted-by":"publisher","first-page":"5378","DOI":"10.1021\/acs.jproteome.5b00932","article-title":"Parallel accumulation-serial\nfragmentation\n(PASEF): multiplying sequencing speed and sensitivity by synchronized\nscans in a trapped ion mobility device","volume":"14","author":"Meier","year":"2015","journal-title":"J. Proteome\nRes."},{"issue":"16","key":"2026093015110114200_cit26","doi-asserted-by":"publisher","first-page":"6139","DOI":"10.1021\/acs.analchem.1c05090","article-title":"Coupling Stable\nIsotope Labeling and Liquid Chromatography-Trapped\nIon Mobility Spectrometry-Time-of-Flight-Tandem Mass Spectrometry\nfor Mosquito Ovarian Lipid Studies","volume":"94","author":"Tose","year":"2022","journal-title":"Anal. Chem."},{"key":"2026093015110114200_cit27","doi-asserted-by":"publisher","first-page":"100485","DOI":"10.1016\/j.forc.2023.100485","article-title":"Application\nof mass spectrometry fragmentation patterns for rapid screening and\nstructure identification of fentanyl analogues in suspicious powder","volume":"33","author":"Fan","year":"2023","journal-title":"Forensic Chem."},{"key":"2026093015110114200_cit28","doi-asserted-by":"publisher","first-page":"496","DOI":"10.1002\/dta.2758","article-title":"The characterization of isobaric\nproduct ions of fentanyl using multi-stage mass spectrometry, high-resolution\nmass spectrometry and isotopic labeling","volume":"12","author":"Davidson","year":"2020","journal-title":"Drug\nTest Anal."},{"key":"2026093015110114200_cit29","doi-asserted-by":"publisher","first-page":"117185","DOI":"10.1016\/j.ijms.2023.117185","article-title":"Determining\nprotonation site in fentanyl protomers\nusing ion mobility-aligned MS\/MS fragmentation","volume":"496","author":"Aderorho","year":"2024","journal-title":"Int. J. Mass Spectrom."},{"issue":"4","key":"2026093015110114200_cit30","doi-asserted-by":"publisher","first-page":"369","DOI":"10.1002\/dta.3550","article-title":"Improved separation of fentanyl isomers using metal\ncation adducts and high-resolution ion mobility-mass spectrometry","volume":"16","author":"Aderorho","year":"2024","journal-title":"Drug Test. Anal."},{"issue":"10","key":"2026093015110114200_cit31","doi-asserted-by":"publisher","first-page":"2323","DOI":"10.1039\/C8AN00138C","article-title":"Metal ions induced secondary structure\nrearrangements: mechanically interlocked lasso vs. unthreaded branched-cyclic\ntopoisomers","volume":"143","author":"Jeanne Dit Fouque","year":"2018","journal-title":"Analyst"},{"issue":"37","key":"2026093015110114200_cit32","doi-asserted-by":"publisher","first-page":"8869","DOI":"10.1021\/acs.jpcb.4c02696","article-title":"Gas-Phase\nStructures of Fucosylated Oligosaccharides: Alkali Metal and Halogen\nInfluences","volume":"128","author":"Miller","year":"2024","journal-title":"J. Phys. Chem. B"},{"issue":"8","key":"2026093015110114200_cit33","doi-asserted-by":"publisher","DOI":"10.1002\/jms.5070","article-title":"Dependency\nof fentanyl analogue protomer ratios on solvent conditions as measured\nby ion mobility-mass spectrometry","volume":"59","author":"Johnson","year":"2024","journal-title":"J. Mass Spectrom."},{"issue":"5","key":"2026093015110114200_cit34","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1093\/jat\/30.5.335","article-title":"Analysis of 13 Fentanils,\nIncluding Sufentanil and\nCarfentanil, in Human Urine by Liquid Chromatography-Atmospheric-Pressure\nIonization-Tandem Mass Spectrometry","volume":"30","author":"Wang","year":"2006","journal-title":"J. Anal.\nToxicol."},{"key":"2026093015110114200_cit35","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1016\/j.jchromb.2014.05.025","article-title":"Comparison\nof two automated solid phase extractions for the detection of ten\nfentanyl analogs and metabolites in human urine using liquid chromatography\ntandem mass spectrometry","volume":"962","author":"Shaner","year":"2014","journal-title":"J. Chromatogr. B"},{"issue":"1","key":"2026093015110114200_cit36","doi-asserted-by":"publisher","first-page":"36","DOI":"10.23907\/2017.004","article-title":"Emerging\nSynthetic Fentanyl Analogs","volume":"7","author":"Schueler","year":"2017","journal-title":"Acad. Forensic Pathol."}],"container-title":["Journal of the American\nSociety for Mass Spectrometry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jasms.5c00203","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jamsef\/article-pdf\/36\/10\/2206\/40773865\/js5c00203.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jamsef\/article-pdf\/36\/10\/2206\/40773865\/js5c00203.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T19:15:26Z","timestamp":1790795726000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jamsef\/article\/36\/10\/2206\/3651182\/Complementary-Separation-of-Novel-Synthetic"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,10]]},"references-count":36,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,9,10]]},"published-print":{"date-parts":[[2025,10,1]]}},"URL":"https:\/\/doi.org\/10.1021\/jasms.5c00203","relation":{},"ISSN":["1044-0305","1879-1123"],"issn-type":[{"value":"1044-0305","type":"print"},{"value":"1879-1123","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,10]]}}}