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Although urinary 2,5\u2010HD is widely used for biological monitoring, its interpretation remains complicated by analytical heterogeneity, particularly the distinction between free 2,5\u2010HD and acid\u2010hydrolyzed total 2,5\u2010HD. This review integrates mechanistic toxicology and analytical biomonitoring evidence to clarify how different biomarkers reflect different stages of n\u2010hexane neurotoxicity. 2,5\u2010HD reacts with lysine residues to form pyrrole adducts, promotes neurofilament cross\u2010linking, disrupts axonal transport, and contributes to distal axonal degeneration. Urinary 2,5\u2010HD is an established occupational exposure biomarker, although free and acid\u2010hydrolyzed total 2,5\u2010HD are not interchangeable. Pyrrole adducts are candidate toxicodynamic biomarkers supported predominantly by animal studies and limited worker evidence. Neurofilament light chain (Nf\u2010L) is an exploratory, nonspecific axonal\u2010injury biomarker with 2,5\u2010HD evidence confined to animal models and no human occupational validation for n\u2010hexane. Accordingly, we present the three analytes as a research framework with unequal validation maturity rather than as co\u2010equal, clinically actionable layers. Future longitudinal occupational studies should evaluate them together with air monitoring, electrophysiology, and clinical outcomes to determine whether the candidate pyrrole\u2010adduct and exploratory Nf\u2010L layers add temporal or practical value beyond established exposure monitoring.<\/jats:p>","DOI":"10.1002\/jat.70429","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T06:00:09Z","timestamp":1788760809000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An Evidence\u2010Based Biomarker Framework for n\u2010Hexane\u2010Induced Neuropathy: Urinary 2,5\u2010Hexanedione, Pyrrole Adducts, and Neurofilament Light Chain"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-9447-327X","authenticated-orcid":false,"given":"Bahad\u0131r","family":"Aliko\u00e7","sequence":"first","affiliation":[{"name":"Department of Medical Biochemistry Ankara Bilkent City Hospital  Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9630-3852","authenticated-orcid":false,"given":"G\u00fclsen","family":"Y\u0131lmaz","sequence":"additional","affiliation":[{"name":"Department of Medical Biochemistry Ankara Bilkent City Hospital  Ankara T\u00fcrkiye"},{"name":"Ankara Y\u0131ld\u0131r\u0131m Beyaz\u0131t University  Ankara T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,6]]},"reference":[{"key":"e_1_2_16_2_1","doi-asserted-by":"publisher","DOI":"10.1136\/oem.33.2.92"},{"key":"e_1_2_16_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/0041\u2010008X(85)90114\u20100"},{"key":"e_1_2_16_4_1","doi-asserted-by":"publisher","DOI":"10.1155\/2014\/239240"},{"key":"e_1_2_16_5_1","volume-title":"Toxicological Profile for N\u2010Hexane","author":"Agency for Toxic Substances and Disease Registry","year":"2025"},{"key":"e_1_2_16_6_1","doi-asserted-by":"publisher","DOI":"10.1136\/oem.45.2.133"},{"key":"e_1_2_16_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010020\u201076990\u20107"},{"key":"e_1_2_16_8_1","doi-asserted-by":"publisher","DOI":"10.1136\/bmjno\u20102021\u2010000143"},{"key":"e_1_2_16_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells11162499"},{"key":"e_1_2_16_10_1","doi-asserted-by":"publisher","DOI":"10.1539\/joh.41.95"},{"key":"e_1_2_16_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022\u2010510X(82)90028\u20104"},{"key":"e_1_2_16_12_1","volume-title":"Threshold Limit Values for Chemical Substances and Physical Agents & Biological Exposure Indices","author":"American Conference of Governmental Industrial Hygienists","year":"2026"},{"key":"e_1_2_16_13_1","doi-asserted-by":"publisher","DOI":"10.1097\/00005072\u2010198309000\u201000007"},{"key":"e_1_2_16_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/acn3.51234"},{"key":"e_1_2_16_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF03028034"},{"key":"e_1_2_16_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00586062"},{"key":"e_1_2_16_17_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00381240"},{"key":"e_1_2_16_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac800065d"},{"key":"e_1_2_16_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0021\u20109673(92)85092\u20108"},{"key":"e_1_2_16_20_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.pa.22.040182.001045"},{"key":"e_1_2_16_21_1","doi-asserted-by":"publisher","DOI":"10.3390\/foods11213412"},{"key":"e_1_2_16_22_1","doi-asserted-by":"publisher","DOI":"10.1177\/096032719701600301"},{"key":"e_1_2_16_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0009\u20102797(85)80167\u20108"},{"key":"e_1_2_16_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/0041\u2010008x(85)90287\u2010x"},{"key":"e_1_2_16_25_1","volume-title":"MAK\u2010 und BAT\u2010Werte\u2010Liste 2025: St\u00e4ndige Senatskommission zur Pr\u00fcfung gesundheitssch\u00e4dlicher Arbeitsstoffe Mitteilung 61","author":"Deutsche Forschungsgemeinschaft","year":"2025"},{"key":"e_1_2_16_26_1","volume-title":"2,5\u2010Hexanedione in Urine: Back\u2010Up Data Report. 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