{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T07:26:39Z","timestamp":1786692399827,"version":"build-2736575974"},"reference-count":27,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["K08AI152942"],"award-info":[{"award-number":["K08AI152942"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000025","name":"National Institute of Mental Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000025","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["UM1AI068632"],"award-info":[{"award-number":["UM1AI068632"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["UM1AI068616"],"award-info":[{"award-number":["UM1AI068616"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["UM1AI106716"],"award-info":[{"award-number":["UM1AI106716"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biomedical Chromatography"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Isoniazid (INH) is a key component of tuberculosis treatment regimens but is also associated with hepatotoxicity. This toxicity is mediated in part by its metabolites, but some degrade rapidly in plasma. A rapid freeze\/thaw process with a methanol protein precipitation extraction was developed to slow the rapid degradation of acetylisoniazid (AcINH) into isonicotinic acid (INA) and acetylhydrazine (AcHZ) in human plasma. An assay consisting of two UHPLC\u2013MS\/MS methods was developed and validated to determine concentrations of INH and four of its metabolites in human plasma: the \u201cpolar\u201d method covering AcINH and INA, and the \u201cnonpolar\u201d method covering p\u2010tolualdehyde derivatized AcHZ, hydrazine (HZ), and INH. Polar and nonpolar method chromatographic separation was accomplished with biphenyl or C18 columns, respectively, mobile phases of 4\u2009mM ammonium formate in water or acetonitrile, and method specific gradient conditions. The validated range for AcINH, INA, and INH was 10.0 to 5000\u2009ng\/mL and for AcHZ and HZ was 10.0 to 500\u2009ng\/mL from 0.1\u2009mL of plasma, respectively. Though other groups have validated methods to quantitate some or all of these analytes, none have adequately characterized or addressed AcINH degradation. This validated assay will help facilitate a better understanding of INH metabolite disposition and relationships with toxicity.<\/jats:p>","DOI":"10.1002\/bmc.70550","type":"journal-article","created":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T10:55:34Z","timestamp":1784285734000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Validation of an UHPLC\u2013MS\/MS Assay for the Quantification of Isoniazid and Four Major Metabolites in Human Plasma"],"prefix":"10.1002","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-1222-1590","authenticated-orcid":false,"given":"David","family":"Nerguizian","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences Skaggs School of Pharmacy and Pharmaceutical Sciences University of Colorado Anschutz Medical Campus  Aurora Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brandon","family":"Klein","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences Skaggs School of Pharmacy and Pharmaceutical Sciences University of Colorado Anschutz Medical Campus  Aurora Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farah","family":"Abdelmawla","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences Skaggs School of Pharmacy and Pharmaceutical Sciences University of Colorado Anschutz Medical Campus  Aurora Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joe","family":"Gomez","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences Skaggs School of Pharmacy and Pharmaceutical Sciences University of Colorado Anschutz Medical Campus  Aurora Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter L.","family":"Anderson","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences Skaggs School of Pharmacy and Pharmaceutical Sciences University of Colorado Anschutz Medical Campus  Aurora Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lane","family":"Bushman","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences Skaggs School of Pharmacy and Pharmaceutical Sciences University of Colorado Anschutz Medical Campus  Aurora Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3832-8699","authenticated-orcid":false,"given":"Kristina M.","family":"Brooks","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences Skaggs School of Pharmacy and Pharmaceutical Sciences University of Colorado Anschutz Medical Campus  Aurora Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,17]]},"reference":[{"key":"e_1_2_7_2_1","volume-title":"StatPearls","author":"Badrinath M.","year":"2026"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/ardp.70204"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1177\/096032718500400210"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.2100"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.00655\u20102019"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF01740658"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1080\/10715760400008854"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1813060"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2014.09.019"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2008.12.024"},{"key":"e_1_2_7_12_1","volume-title":"ICH M10: Bioanalytical Method Validation","author":"International Council for Harmonisation (ICH)","year":"2022"},{"key":"e_1_2_7_13_1","unstructured":"Klein B. Z.Wei D.Nerguizian et\u00a0al.2024. \u201cPharmacokinetics of Isoniazid Metabolites During Pregnancy and Postpartum [CROI Abstract 871].\u201d Abstracts from the CROI 2024 Conference on Retroviruses and Opportunistic Infections. 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