PT04 Endocrine-Disrupting Chemicals in Wastewater Treatment Effluents
Exhibit Hall
8:00 AM - Tuesday

(PT046) Identification and evaluation of hormonally-active compounds from chemical recovery condensates at a Canadian pulp and paper mill.

Belknap, A1, 3, Shaughnessy, K2, MacLatchy, D2, Solomon, K1, Hewitt, M3, 1 Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada3 National Water Research Institute, Environment Canada, Burlington, Ontario, Canada2 Department of Biology, University of New Brunswick, Saint John, New Brunswick, Canada

ABSTRACT- There is considerable evidence that kraft pulp and paper mill recovery condensates are a primary source of hormonally active compounds to fish. 5th effect softwood chemical recovery condensates have been collected from an Eastern Canadian pulp and paper mill and were extracted using an established solid phase extraction (SPE) protocol. In a series of whole fish bioassays, testosterone concentrations were measured in blood plasma following a 7-day static exposure of.mummichog (Fundulus heteroclitus). Fish were exposed to a 5-point dilution series of condensate SPE extract to establish an effective dose for ongoing toxicity identification evaluations (TIE). A significant depression in plasma testosterone was observed relative to a laboratory blank in the highest extract exposure concentration (4% v/v) only. A preparative high pressure liquid chromatography fractionation method for softwood condensate extracts was developed and fish were exposed to 4 fractions. Active fractions were subjected to a second round of HPLC subfractionation and analyzed by GC-MS. Unique constituents were identified by retention time and mass spectral fragmentation patterns. A plant based diterpene, 13-epimanool was confirmed in the condensate extracts and is being used as a reference standard for analytical fractionation recoveries and as a surrogate to estimate condensate potency. HPLC fractions and subfractions were screened by GC-MS in SIM mode for identification and quantification of manool (m/z = 55, 69, 81, 137) relative to a calibartion standard. The results presented here demonstrate a linkage between the hormonal activity associated with kraft condensates and the presence of polycyclic aromatic hydrocarbon-based extractives.

Key words: pulp mill effluent, mummichog, 13-epimanool, TIE

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