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To understand potential biological effects of degradation intermediates, we tested three fractions of DOC: (1) solid-phase extract (HLB); (2) dichloromethane (DCM-total) extract used in TPHd; and (3) DCM extract with hydrocarbons isolated by silica gel cleanup (DCM-SGC). Bioactivity of extracts from five wells spanning a range of DOC was tested using an in vitro multiplex reporter system that evaluates modulation of the activity of 46 transcription factors; extracts were evaluated at concentrations equivalent to the well water samples. The aryl hydrocarbon receptor (AhR) and pregnane X receptor (PXR) transcription factors showed the greatest upregulation, with HLB exceeding DCM-total, and no upregulation in the hydrocarbon fraction (DCM-SGC). The HLB extracts were further studied with HepG2 chemically activated luciferase expression (CALUX) in vitro assays at nine concentrations ranging from 40 to 0.01 times the well water concentrations. Responses decreased with distance from the source but were still present at two wells without detectable hydrocarbons. Thus, our in vitro assay results indicate that risks associated with degradation intermediates of hydrocarbons in groundwater will be underestimated when protocols that remove these chemicals are employed.<\/jats:p>","DOI":"10.1021\/acs.est.0c02220","type":"journal-article","created":{"date-parts":[[2020,8,13]],"date-time":"2020-08-13T15:01:09Z","timestamp":1597330869000},"page":"11396-11404","source":"Crossref","is-referenced-by-count":37,"title":["Biological\nEffects of Hydrocarbon Degradation Intermediates:\nIs the Total Petroleum Hydrocarbon Analytical Method Adequate for\nRisk Assessment?"],"prefix":"10.1021","volume":"54","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1411-6018","authenticated-orcid":true,"given":"Barbara A.","family":"Bekins","sequence":"first","affiliation":[{"name":"USGS , , ,","place":["Menlo Park, California, United States, 94025"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0386-3496","authenticated-orcid":true,"given":"Jennifer C.","family":"Brennan","sequence":"additional","affiliation":[{"name":"USGS , , ,","place":["Columbia, Missouri, United States, 65201"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donald E.","family":"Tillitt","sequence":"additional","affiliation":[{"name":"USGS , , ,","place":["Columbia, Missouri, United States, 65201"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5123-1007","authenticated-orcid":true,"given":"Isabelle M.","family":"Cozzarelli","sequence":"additional","affiliation":[{"name":"USGS , , ,","place":["Reston, Virginia, United States, 20192"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer McGuire","family":"Illig","sequence":"additional","affiliation":[{"name":"University of St. Thomas , , ,","place":["St. Paul, Minnesota, United States, 55105"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dalma","family":"Martinovic\u0301-Weigelt","sequence":"additional","affiliation":[{"name":"University of St. Thomas , , ,","place":["St. Paul, Minnesota, United States, 55105"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,8,13]]},"reference":[{"key":"2026090303325249600_cit1","first-page":"292","volume-title":"Natural Attenuation for Groundwater Remediation","author":"NRC.","year":"2000"},{"key":"2026090303325249600_cit2","first-page":"422","volume-title":"Alternatives for Managing the Nation\u2019s Complex Contaminated\nGroundwater Sites","author":"NRC.","year":"2013"},{"key":"2026090303325249600_cit3","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1007\/BF01890379","article-title":"Transformation of monoaromatic hydrocarbons\nto organic acids in anoxic groundwater environment","volume":"16","author":"Cozzarelli","year":"1990","journal-title":"Environ. 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