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Abstract
The importance of sample preparation techniques cannot be overstated as the accurate sample preparation is essential. Often overlooked, it is the midway point where the analytes from the sample matrix are transformed so they are suitable for analysis. Even the best analytical techniques cannot rectify problems generated by sloppy sample pre-treatment. In the present work effectiveness of six different commonly applied extraction techniques for the determination of robenidine in poultry feed has been compared. The result analysis was performed using non-parametric Kruskal-Wallis (K-W) test and cluster analysis. The extraction performance of shaking, Soxhlet, Soxtec, ultrasonically assisted extraction (UAE), microwave–assisted extraction (MAE) and accelerated solvent extraction (ASE) was investigated. Operation parameters such as extraction pressure, extraction temperature and extraction time were evaluated as the variable parameters under the experimental ranges of 1000–2500 psi, 60–140°C and from 1 minute up to 6 hours, respectively. The effect of operation parameters during extraction on the recovery of robenidine from poultry feed, reproducibility of the methods and solvent consumption were investigated using spiked samples. Every single extract was subjected to clean–up using aluminium oxide column (Pasteur pipette filled with 1 g of aluminium oxide) from which robenidine was eluted with 10 ml of methanol. The eluate from the clean-up column was collected in a volumetric flask and analyzed by HPLC–DAD–MS technique. In general, all extraction techniques allowed the isolation of robenidine from poultry feed, but the recovery obtained by means of modern extraction techniques (ASE, MAE) was higher than that obtained with conventional techniques (Soxhlet, UAE). Also, RSD showed to be lower for novel extraction techniques.