title: Investigations of cholesterol transformation during sewage treatment: relevance to estrogen formation pathways?
abstract: There is currently a great deal of concern over the observation of so-called estrogenic effects (specifically increases in the concentrations of the egg yolk precursor, vitellogenin) in male fish living in some UK rivers. The effects have been attributed to chemicals, including estrogenic steroids, which enter the rivers via sewage effluents. The origins of these estrogenic steroids in sewage may include contributions from the influents and possibly in situ transformation processes of other steroids occurring during sewage treatment. The present study examined the latter possibility. The bacterial metabolism of radiolabelled cholesterol during laboratory-simulated aerobic sewage treatment was studied by reverse phase radio-high performance liquid chromatography (rHPLC) and radio-gas chromatography (rGC) to examine the hypothesis that cholesterol could undergo A-ring aromatisation to form first, 19-norcholest-1,3,5(10)-trien-3-ol (NCT) and hence, by known bacterial pathways, the estrogenic steroid, estrone. The results showed that, to the contrary, much of the cholesterol (approx. 50% in 96 h) underwent rapid mineralisation to carbon dioxide, consistent with A-ring rupture (rather than aromatisation) and beta-oxidation of the alkyl side chain as major transformation routes. Some polar (early-eluting) rHPLC products were observed, possibly steroidal conjugates and/or fatty acids. Among the minor metabolites detected by rGC and GC-mass spectrometry (GC-MS) were cholest-3,5-diene and a second cholestadiene isomer. However, since alkenes were unexpected in this rHPLC fraction, they may arise as artefacts from thermal decomposition of cholesteryl esters, indicating that some cholesterol esterification had also occurred. In the alcohol rHPLC fractions, cholestadienol was identified by GC and GC-MS but neither NCT or estrone were detected. This suggests that, at least under these simulated conditions, in situ aromatisation of cholesterol to NCT and formation of estrone from NCT were not major processes.
authors.affiliation: Department of Environmental Sciences, Plymouth Environmental Research Centre, University of Plymouth, UK.
authors.forename: S J
authors.initials: SJ
authors.lastname: Niven
authors.forename: J
authors.initials: J
authors.lastname: Snape
authors.forename: M
authors.initials: M
authors.lastname: Hetheridge
authors.forename: M
authors.initials: M
authors.lastname: Evans
authors.forename: J
authors.initials: J
authors.lastname: McEvoy
authors.forename: S J
authors.initials: SJ
authors.lastname: Rowland
mesh_terms: D000818:Animals; D001711:Biotransformation; D002245:Carbon Dioxide / Q000032:analysis; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004781:Environmental Exposure; D004967:Estrogens / Q000737:chemistry; D005399:Fishes; D008401:Gas Chromatography-Mass Spectrometry; D006854:Hydrocortisone / Q000031:analogs & derivatives* / Q000737:chemistry; D010084:Oxidation-Reduction; D012722:Sewage / Q000382:microbiology*; D014819:Vitellogenins / Q000032:analysis
chemical_list: D004967:Estrogens; D012722:Sewage; D014819:Vitellogenins; D002245:Carbon Dioxide; C027515:19-norcortisol; D002784:Cholesterol; D006854:Hydrocortisone