Chemical Extraction of Pesticide-Treated Carrot Callus to Assess Cytotoxicity of the Pesticide Metabolites

To begin, mix 0.2 grams of ground powder of each freeze dried, pesticide-treated carrot callous sample, with three milliliters of acetonitrile in a 10 milliliter centrifuge tube. Vortex the centrifuge tube for eight minutes, and then sonicate it for five minutes at 150 watts and 40 kilohertz. Next, centrifuge the tube at 8, 000G and four degrees Celsius for 10 minutes, before collecting the supernatant by pipetting.

Using a nitrogen-blowing concentrator at 40 degrees Celsius, concentrate the pooled extracts to dryness. Redissolve the residues of the extract in one milliliter of DMEM, containing 0.3%DMSO to yield the pesticide metabolite exposure solution. In a similar manner, redissolve the residues of the extract obtained from the blank callous samples.

in one milliliter of DMEM, containing 0.3%DMSO. Then, dissolve one milligram of each pesticide used in different tubes containing the blank sample residue solution to yield apparent pesticide exposure solutions.