Copper-dependent hydrolysis of trichloronate by turkey serum studied with use of new analytical procedure based on application of chiral chromatography and UV/Vis spectrophotometry

Abstract Trichloronate is a racemic organophosphate, which has been used for the manufacture of insecticides. This compound induces delayed neuropathy in hen and humans. This study shows the Cu 2+ -dependent hydrolysis of trichloronate by turkey serum using UV/Vis spectrophotometry and chiral chromatography. The CHIRALCEL OD column and mobile phase of heptane allowed a resolution of 1.15 of its two enantiomers, while the liquid-liquid extraction showed a recovery of 95–98%. The optimum linear response was of 50 to 800 μM with a detection and quantification limit of 0.6 and 2 μM for (+)-trichloronate, and 0.7 and 2.3 μM for (−)-trichloronate. The levels of Cu 2+ -dependent hydrolysis (μM remaining concentration) quantified for 60 min at 37 °C and pH 7.4 were statistically higher (p ˂ 0.05) for (−)-trichloronate (65%) than (+)-trichloronate (32%). This stereoselective hydrolysis was confirmed by UV/Vis spectrophotometry using 2,4,5‑trichlorophenol as standard, each of the enantiomers (93–95% purity) collected by HPLC, as well as aminoantipyrine and ferricyanide reagents to yield a colored product. This method exhibited an optimal linearity ( r  > 0.99) and a higher Cu 2+ -dependent hydrolysis (p  2+ -dependent stereoselective hydrolysis of a racemic OP in its thio form (P = S) by an A-esterase of the turkey serum through the development of a colorimetric method and optimization of an existing chiral chromatographic method.