Development and application of a mini-QuEChERS method for the determination of pesticide residues in anuran adipose tissues

Abstract

The expansion of monocultures to regions close to conservation areas has put biodiversity at risk, mainly due to the intense use of pesticides. Anurans are highly susceptible to pesticides and may be a biological marker in the contamination of an area. However, methods for determining pesticides in anurans are incipient. In this work, a miniaturized QuEChERS method was developed for the extraction of atrazine, chlorpyrifos, α- and β-endosulfan, α-, β-, θ- and ζ-cypermethrin in anuran adipose tissues. The method was optimized for the tissue sample size scale according to sample mass availability. Extracting solvent and adsorbents for the clean-up step was evaluated, achieving recoveries next to 100% with acetonitrile and without a clean-up step. The mini-QuEChERS method, using 500 mg of adipose tissue, 50 mg of NaCl and 200 mg of MgSO₄, 100 μL of ultrapure water, and 1.50 mL of acetonitrile with no purification step, followed by high-performance liquid chromatography analysis and photodiode array detection was validated following the European Community guidelines. The methodology showed a moderate matrix effect for some pesticides, which was corrected using the matrix-matched calibration. The limits of quantification for the pesticide residues in adipose tissues ranged from 10 to 75 μg kg⁻¹. Pesticide recoveries ranged from 74% to 115%, and repeatability and within-lab reproducibility showed relative standard deviations < 11%. The mini-QuEChERS method was applied to extract pesticide residues from the adipose tissues of two species of anurans: Leptodactylus macrosternum and Scinax x-signatus. 25% of samples were positive, detecting endosulfan and chlorpyriphos, confirmed by liquid chromatography coupled to tandem mass spectrometry. The mini-QuEChERS was a simple, economical, and eco-friendly method for extracting pesticide residues in anuran adipose tissue samples.