{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T15:21:23Z","timestamp":1788448883509,"version":"build-2803163510"},"reference-count":43,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000066","name":"National Institute of Environmental Health Sciences","doi-asserted-by":"publisher","award":["R21ES034457"],"award-info":[{"award-number":["R21ES034457"]}],"id":[{"id":"10.13039\/100000066","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000066","name":"National Institute of Environmental Health Sciences","doi-asserted-by":"publisher","award":["R56ES013527"],"award-info":[{"award-number":["R56ES013527"]}],"id":[{"id":"10.13039\/100000066","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000071","name":"National Institute of Child Health and Human Development","doi-asserted-by":"publisher","award":["R01HD108165"],"award-info":[{"award-number":["R01HD108165"]}],"id":[{"id":"10.13039\/100000071","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Chem."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Biomonitoring of commercial endocrine-disrupting chemicals (EDCs) increasingly underpins exposure assessment and risk evaluation, yet most large-scale studies still rely on indirect, enzymatic hydrolysis\u2013based methods whose quantitative performance has not been systematically verified. Here, we describe the development and rigorous validation of a direct liquid chromatography-tandem mass spectrometry (LC-MS\/MS) assay using validated standards to simultaneously quantify bisphenol S (BPS), propylparaben (PrP), monobutyl phthalate (MBP), and their major urinary glucuronide and sulfate conjugates in human urine.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A direct LC\u2013MS\/MS assay was developed for the simultaneous quantification of BPS, PrP, MBP and their major urinary metabolites. The method was validated in accordance with U.S. Food and Drug Administration bioanalytical guidelines, including assessments of linearity, accuracy, precision, selectivity, specificity, matrix effects, recovery, and carryover. The validated assay was then used to evaluate the accuracy of a conventional \u03b2\u2010glucuronidase\u2013based hydrolysis workflow across a wide concentration range in spiked synthetic urine by comparing indirect measurements with direct totals for each analyte. Method utility in human biomonitoring was demonstrated by analysis of urine samples from 30 pregnant women in their second trimester.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Using isotope-dilution calibration, solid-phase extraction, and negative-ion electrospray multiple reaction monitoring, the method achieved sub\u2010ng\/mL limits of detection for all analytes, linear response over 3-4 orders of magnitude, and intra\u2010 and inter\u2010day precision and accuracy within contemporary bioanalytical criteria, confirming fitness for trace-level biomonitoring. Indirect, hydrolysis-based measurements closely tracked direct totals for BPS and PrP, but systematically underestimated MBP, with a concentration-dependent negative bias that increased at higher levels, demonstrating that hydrolysis efficiency is analyte-specific and cannot be inferred from surrogate substrates alone. Application of the direct method to archived urine samples from 30 pregnant individuals enabled the first simultaneous resolution of the free and conjugated forms of these three EDCs in a maternal cohort and revealed that glucuronides accounted for the majority of the total urinary burden, with total concentrations exceeding contemporary NHANES estimates.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>Collectively, these findings show that indirect methods can introduce substantial, analyte-dependent underestimation of internal dose, with implications for exposure misclassification, attenuation of epidemiologic effect estimates, and underestimation of population risk. The data support the position that direct LC\u2013MS\/MS quantification of parent and conjugated species, coupled with analyte-resolved assessment of hydrolysis efficiency, should become standard practice in method validation and national biomonitoring programs to ensure accurate exposure assessment for non\u2010persistent EDCs.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fchem.2026.1894102","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T14:53:13Z","timestamp":1788447193000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Biomonitoring: a demonstration of the importance of direct analytical methods for the assessment of chemical exposures"],"prefix":"10.3389","volume":"14","author":[{"given":"Roy","family":"Gerona","sequence":"first","affiliation":[{"name":"Department of Obstetrics, Gynecology and Reproductive Sciences, Center for Reproductive Sciences, University of California, San Francisco","place":["San Francisco, CA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alia","family":"Sovereign","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, Gynecology and Reproductive Sciences, Center for Reproductive Sciences, University of California, San Francisco","place":["San Francisco, CA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"Hunt","sequence":"additional","affiliation":[{"name":"School of Molecular Biosciences and Center for Reproductive Biology, Washington State University","place":["Pullman, WA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"568","DOI":"10.2133\/dmpk.dmpk-10-rg-013","article-title":"Metabolism of parabens (4-hydroxybenzoic acid esters) by hepatic esterases and UDP-Glucuronosyltransferases in man","volume":"25","author":"Abbas","year":"2010","journal-title":"Drug Metab. 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