engleski

Organochlorine insecticides in selected vegetable and fruit samples from Zagreb open market and their cytotoxic effects on BHK (Baby Hamster Kidney) 21 C13 cells

Selected vegetable (carrott, parsley root and leaf, leek root and leaf, tomato) and fruit (apple peel and core, strawberry, banana, kiwi, pear peel and core) samples from the open market of Zagreb were analysed on the possible presence of organochlorine insecticides. By gas chromatographic analysis the presence of lindane, aldrin, heptachlor and alpha, beta, delta-HCH were detected. The concentrations found in the tested samples were around 2 to 10 times lower than the permitted levels according to the offical Croatian regulations. In order to reduce number of animals in performing the risk assessment from different chemicals present in the trace concentrations in food, different primary cells or commercial cell lines are introduced for toxicity assays. In the present investigation the toxicity of lindane, aldrin and heptachlor were determined in BHK (Baby Hamster Kidney)21 C13 cell line. Following parameters were examined: cell viability and number of cells measured by trypane-blue exclusion method in Fuchs-Rosenthal heamocytometer and lyzosomal activity measured by neutral red method. BHK21 C13 cell line was obtained from the American Type Culture Collection (CCL 10). The cells were maintained in Glasgow BHK 21 medium supplemented with 5% fetal bovine serum in an athmosphere of 95% air and 5% CO_2 at 37 ^0 C. The initial cell concentration was 2-2.3 x 105 cells/mL/well on the 24-Multiwell plates, and cells were seeded 24 hours before the addition of toxicant (lindane, aldrin and heptachlor). Lindane and heptachlor were added in the concentrations of 5 to 100 microg/mL/well, aldrin from 2 to 40 microg/mL/well and the cytotoxicity was determined after 24, 48 and 72 hours. The IC_20, IC_50 and IC_80 values were detected after 72 hours from the slope of % inhibition vs. log dose values. The increasing concentrations within one toxicant resulted in the increasing cytotoxic effect of the tested compounds in the rate as followed: aldrin>lindane>heptachlor.

gas chromatography; organochlorine insecticides; vegetable; fruit; cytotoxicity; BHK21 C13 cell line

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