Alternative Green Preconcentration Approach Based on Ultrasound-Assisted Surfactant-Enhanced Emulsification Microextraction and HPLC for Determination of Benzimidazole Anthelmintics in Milk Formulae

An alternative green microextraction method based on ultrasound-assisted surfactant-enhanced emulsification microextraction (UASEME) using a low-density extraction solvent coupled with HPLC has been developed for preconcentration and determination of six benzimidazole anthelmintics, namely, oxfendazole, albendazole, mebendazole, flubendazole, fenbendazole, and niclosamide. The separation was achieved within 12 min, using an Inertsil(A (R)) C18 column (4.6 x 150 mm, 5.0 A mu m), with a gradient mobile phase of acetonitrile and 0.1 % (v/v) formic acid. Under the optimum UASEME conditions using Tergitol(A (R)) TMN-6 and 1-octanol as emulsifier and extraction solvent, respectively, linearity was in the range of 0.5-5,000 mu g L-1 with the coefficients of determination (R (2)) ranging from 0.9959 to 0.9999. Enrichment factors were obtained up to 89, corresponding to limits of detection ranging from 0.50 to 6.00 A mu g L-1. Intra-day (n = 8) and inter-day (n = 3 x 3) precisions were obtained with relative standard deviations for retention time and peak area of lower than 2 and 15 %, respectively. The proposed method was successfully applied to determine the target benzimidazoles in milk formulae.