In crude oil contaminant plumes,
the dissolved organic carbon (DOC)
is mainly hydrocarbon degradation intermediates only partly quantified
by the diesel range total petroleum hydrocarbon (TPHd) method. 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.