title: Impact of protein binding on the availability and cytotoxic potency of organochlorine pesticides and chlorophenols in vitro.
abstract: In vitro toxicity data are generally based on nominal concentrations and thus depend on both activity and availability of a compound. The aims of the present study were to examine the influence of protein binding on the cytotoxicity of selected organochlorine pesticides and chlorophenols in Balb/c 3T3 cell cultures and to determine parameters of protein binding which can be used to estimate protein bound fractions and to model distribution in vitro. EC(50)-values derived from concentration-effect relationships determined in the presence of various concentrations of bovine serum albumin (BSA) were linearly correlated to BSA concentration. Increasing the BSA concentration from about 1.2 to 40 mg/ml increased the EC(50)-values by factors between 3.4 and 34.4. Molar ratios of substance bound to albumin ranged from 0.11 to 2.42. Calculated fractions bound to albumin in the normal growth medium were 0.075-0.17 (p,p'-DDT, p,p'-DDE, dieldrin, lindane), 0.09-0.1 (4-mono- and 2,4-dichlorophenol), 0.68 (2,4,5-trichlorphenol) and almost 1.0 (pentachlorophenol). At 40 mg/ml BSA any compound was largely bound to albumin (fractions bound > or = 0.74). Distribution modelling revealed that the availability of the highly hydrophobic organochlorines additionally was significantly reduced by partitioning into lipids. The results clearly demonstrate that nominal and relative toxic potencies of organochlorine pesticides and chlorophenols determined in vitro are substantially influenced by effects of protein binding on availability.
authors.affiliation: Institut für Experimentelle Toxikologie, Universitätsklinikum Kiel, Weimarer Str. 8, Haus 3, D-24106 Kiel, Germany. guelden@toxi.uni-kiel.de
authors.forename: Michael
authors.initials: M
authors.lastname: Gülden
authors.forename: Sabine
authors.initials: S
authors.lastname: Mörchel
authors.forename: Sail
authors.initials: S
authors.lastname: Tahan
authors.forename: Hasso
authors.initials: H
authors.lastname: Seibert
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001682:Biological Availability; D002452:Cell Count; D002733:Chlorophenols / Q000493:pharmacokinetics* / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D007306:Insecticides / Q000493:pharmacokinetics* / Q000633:toxicity; D007700:Kinetics; D050356:Lipid Metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008954:Models, Biological; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D002733:Chlorophenols; D007306:Insecticides; D012710:Serum Albumin, Bovine