Determination of Piperazine in Eggs Using Accelerated Solvent Extraction (ASE) and Solid Phase Extraction (SPE) with High-Performance Liquid Chromatography – Fluorescence Detection (HPLC-FLD) and Pre-Column Derivatization with Dansyl Chloride

Abstract

A high-performance liquid chromatography – fluorescence detection (HPLC-FLD) method was established for the determination of piperazine residues in whole eggs, albumen, and yolk employing pre-column derivatization with dansyl chloride. The egg samples were treated by accelerated solvent extraction (ASE) and the analytes purified by solid phase extraction (SPE). Acetonitrile/ultrapure water was used as the mobile phase for gradient elution. The linear dynamic range was from 6.80 to 200.0 µg/kg for whole eggs, 7.50 to 200.0 µg/kg for albumen, and 6.50 to 200.0 µg/kg for yolk with coefficients of determination (R2) greater than or equal to 0.9992. The average recoveries of piperazine from egg samples were from 72.86 to 89.26% when the concentrations were equal to the limit of quantification, 0.5 times the maximum residue limit, equal to the maximum residue limit, and twice the maximum residue limits. The relative standard deviations were between 1.73 and 4.99%. The limits of detection of piperazine in eggs were from 1.92 to 2.50 µg/kg, and the limits of quantification were between 6.50 and 7.50 µg/kg. The method meets the requirements of the European Union, the Ministry of Agriculture of the People’s Republic of China and the U.S. Food and Drug Administration (FDA) for the determination of veterinary drug residues. The validated HPLC-FLD method was successfully applied to the determination of piperazine residues in 50 eggs purchased from local supermarkets.