title: Intra- and interlaboratory reliability of a cryopreserved trout hepatocyte assay for the prediction of chemical bioaccumulation potential.
abstract: Measured rates of intrinsic clearance determined using cryopreserved trout hepatocytes can be extrapolated to the whole animal as a means of improving modeled bioaccumulation predictions for fish. To date, however, the intra- and interlaboratory reliability of this procedure has not been determined. In the present study, three laboratories determined in vitro intrinsic clearance of six reference compounds (benzo[a]pyrene, 4-nonylphenol, di-tert-butyl phenol, fenthion, methoxychlor and o-terphenyl) by conducting substrate depletion experiments with cryopreserved trout hepatocytes from a single source. O-terphenyl was excluded from the final analysis due to nonfirst-order depletion kinetics and significant loss from denatured controls. For the other five compounds, intralaboratory variability (% CV) in measured in vitro intrinsic clearance values ranged from 4.1 to 30%, while interlaboratory variability ranged from 27 to 61%. Predicted bioconcentration factors based on in vitro clearance values exhibited a reduced level of interlaboratory variability (5.3-38% CV). The results of this study demonstrate that cryopreserved trout hepatocytes can be used to reliably obtain in vitro intrinsic clearance of xenobiotics, which provides support for the application of this in vitro method in a weight-of-evidence approach to chemical bioaccumulation assessment.
authors.affiliation: U.S. EPA, ORD, NHEERL, Mid-Continent Ecology Division, Duluth, Minnesota 55804, United States.
authors.forename: Kellie A
authors.initials: KA
authors.lastname: Fay
authors.forename: Robert T
authors.initials: RT
authors.lastname: Mingoia
authors.forename: Ina
authors.initials: I
authors.lastname: Goeritz
authors.forename: Diane L
authors.initials: DL
authors.lastname: Nabb
authors.forename: Alex D
authors.initials: AD
authors.lastname: Hoffman
authors.forename: Barbra D
authors.initials: BD
authors.lastname: Ferrell
authors.forename: Heather M
authors.initials: HM
authors.lastname: Peterson
authors.forename: John W
authors.initials: JW
authors.lastname: Nichols
authors.forename: Helmut
authors.initials: H
authors.lastname: Segner
authors.forename: Xing
authors.initials: X
authors.lastname: Han
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002469:Cell Separation; D002470:Cell Survival; D015925:Cryopreservation; D022781:Hepatocytes / Q000378:metabolism*; D007753:Laboratories; D008297:Male; D017686:Oncorhynchus mykiss / Q000378:metabolism*; D015203:Reproducibility of Results; D015262:Xenobiotics / Q000378:metabolism
chemical_list: D015262:Xenobiotics