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  • Evaluation of in vitro and in vivo toxic effects of newly synthesized benzimidazole-based organophosphorus compounds.

Evaluation of in vitro and in vivo toxic effects of newly synthesized benzimidazole-based organophosphorus compounds.

Ecotoxicology and environmental safety (2012-11-03)
Abbas Güngördü, Nihat Sireci, Hasan Küçükbay, Ayşe Birhanli, Murat Ozmen
ABSTRACT

This paper reports the toxic properties of eight newly synthesized benzimidazole-based organophosphorus (OP) compounds in Xenopus laevis in both in vivo and in vitro conditions. For both experiments, a commercial solution of azinphos methyl (AzM, Gusathion M WP25) was used as a reference compound. The 24-h median lethal concentrations (LC₅₀) of all tested compounds were determined for 46th stage tadpoles in the range of 9.54-140.0 μM. For evaluation of the lethality of the compounds, the activity of the enzyme biomarkers acetylcholinesterase (AChE), carboxylesterase, glutathione S-transferase (GST), glutathione peroxidase, glutathione reductase, lactate dehydrogenase, aspartate aminotransferase, and alanine aminotransferase were determined in vivo in X. laevis tadpoles exposed to three concentrations (LC₅₀, LC₅₀/2, and LC₅₀/4) of tested compounds. All exposure concentrations of AzM and seven of eight tested compounds caused CaE inhibition in in vivo conditions. Furthermore, the AChE inhibition capacity of tested compounds in commercial electric eel AChE and in X. laevis homogenates and also CaE inhibition capacity in only X. laevis homogenates were assayed for a 30-min in vitro exposure period. Eight OP compounds did not inhibit AChE activity more than 23 percent, but AzM exposure inhibited AChE activity by 26 percent for X. laevis homogenates and 97 percent for electric fish AChE in in vitro conditions. Also, CaE inhibition levels in X. laevis tadpole homogenates were 46 percent for AzM and between 8 percent and 33 percent for other compounds in in vitro conditions.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Esterase from Bacillus subtilis, recombinant, expressed in E. coli, ≥10 U/mg
Supelco
Azinphos-methyl, PESTANAL®, analytical standard
Sigma-Aldrich
Esterase from porcine liver, lyophilized, powder, slightly beige, ≥50 U/mg
Sigma-Aldrich
Esterase from rabbit liver, lyophilized powder, ≥30 units/mg protein
Sigma-Aldrich
Esterase from porcine liver, ammonium sulfate suspension, ≥150 units/mg protein (biuret)
Sigma-Aldrich
Esterase from porcine liver, lyophilized powder, ≥15 units/mg solid
Sigma-Aldrich
Esterase Isoenzyme 1 porcine liver, recombinant, recombinant, expressed in E. coli, ≥30.0 U/g
Sigma-Aldrich
Esterase Pseudomonas fluorescens, recombinant from E. coli, ≥4 U/mg
Sigma-Aldrich
Benzimidazole, 98%
Sigma-Aldrich
Esterase from Bacillus stearothermophilus, ≥0.2 U/mg
Sigma-Aldrich
Carboxylesterase 1 isoform b human, recombinant, expressed in baculovirus infected BTI insect cells
Sigma-Aldrich
Carboxylesterase 1 isoform c human, recombinant, expressed in baculovirus infected BTI insect cells
Sigma-Aldrich
Esterase from Bacillus stearothermophilus, recombinant, expressed in E. coli, ≥4.0 U/mg
Sigma-Aldrich
Carboxylesterase 2 human, recombinant, expressed in mouse NSO cells, ≥95% (SDS-PAGE)