IR: ν = 1595 cm−1 (CfN), 1296 cm−1 (CfS), 1087 cm−1 (O–N), 3251 cm−1 (N–H), 3420 cm−1 (O–H).

The 3-(phenylhydrazono) butan-2-one oxime (oxime 2) was prepared by a simple mixture and reflux for 3 h of 1 mol diacetylmonoxime with 1 mol of phenylhydrazine chloride both dissolved in a mixture of ethanol–H2O (2:1, v/v) and 0.5 ml of sodium acetate 6 M. On heating, a dark orange product was formed, collected by filtration, washed with water, and dried in vacuum (yield 70%, mp 190 °C). Pralidoxime (2-PAM; 1-methyl-2-hydroxyiminomethylpyridinium chloride) and Obidoxime (1,3-bis (4-hydroxyiminomethylpyridinium)-2-oxapropane dichloride) JQ1 cell line were purchased from Sigma–Aldrich (Brazil). Chlorpyrifos (O,O-diethyl O-3,5,6-trichloropyridin-2-yl phosphorothioate) was purchased from La Forja S.A. (Montevideo, Uruguay); Diazinon (O,O-diethyl Selleckchem Roxadustat O-[4-methyl-6-(propan-2-yl) pyrimidin-2-yl] phosphorothioate) was purchased from Lusa S.A. (Montevideo, Uruguay) and Malathion

(diethyl 2-[(dimethoxyphosphorothioyl) sulfanyl] butanedioate) was purchased from Nitrosin S.A. (Curitiba, Brazil). All other chemicals used in this study were of analytical purity and were purchased from Sigma–Aldrich (Brazil). Structure of studied oximes and organophosphates is given in Fig. 1. Human blood samples were obtained from healthy volunteers. The blood samples were collected with heparin and then centrifuged for 10 min at 5000 rpm. The plasma was removed as supernatant, 17-DMAG (Alvespimycin) HCl as source of BChE. The erythrocytes were hemolyzed in phosphate buffer (0.1 M, pH7.4) in a ratio 1:10 (w/w), as source of AChE (Jun et al., 2008). The time of enzyme inhibition with the OP was of 1 h. The concentrations of OP used were based on the IC50 values previously experimentally calculated (8.06; 20.72 and 73.00 μM for chlorpyrifos, diazinon and malathion-inhibited AChE, respectively and 1.15; 1.20 and 1.80 μM for chlorpyrifos,

diazinon and malathion-inhibited BChE, respectively). For BChE inhibition, OP solution diluted in phosphate buffer (0.1 M, pH 7.4) was added to the plasma. The same was performed for the inhibition of AChE only that instead of plasma a hemolyzed solution content ethopropazine dichloride 6 mM (to avoid plasma esterase interference) was used. After inhibition, the solution of reactivator (final concentrations of tested reactivators were 1, 10, 50 and 100 μM) in phosphate buffer (0.1 M, pH 7.4) was added to the mixture containing the inhibited enzyme (AChE or BChE). After 10 min of reactivation (Jun et al., 2008), 5,5-dithiobis-2-nitrobenzooic acid (DTNB) in phosphate buffer (0.1 M, pH 7.4) was added and the enzymatic reaction was initiated by addition of acetylthiocholine (ATCh) or butyrylthiocholine (BTCh) substrates. The final concentration of DTNB in the mixture was 0.3 mM and ATCh or BTCh in the mixture was 0.45 mM. The final volume of sample in cuvette was 1 ml.