Automated Accelerated Solvent Extraction Coupled with Ultra-High-performance Liquid Chromatography for the Analysis of Piperazine in Egg Samples

This study involved the development of an ultra-high-performance liquid chromatography-fluorescence method for rapid detection of piperazine residues in eggs. Accelerated solvent extraction, a rapid and efficient sample extraction technique, was used to extract piperazine from egg samples (whole egg, albumin and yolk). Ground eggs were subjected to accelerated solvent extraction using n-hexane in the first step and acetonitrile-formic acid (98:2, v/v) in the second step and were then purified by a solid phase extraction cartridge using polymeric strong cation sorbents. The collected eluate was placed on a nitrogen blower and dried. After reconstitution with acetonitrile, piperazine was derivatized with dansyl chloride and catalytic triethylamine. A portion of the precipitate was removed using a 0.22 μm needle filter (organic-phase) and injected into the separation system for analysis. The analyte was separated using an Acquity HSS T3 [100 × internal diameter (i.d.) 2.1 mm, 1.8 µm] ultra high performance liquid chromatography column that was coupled with a fluorescence detector. Under the optimum conditions, the linear ranges were 3.5–400.0 μg/kg for the whole egg and yolk and 4.2–400.0 μg/kg for the albumin, the extraction recoveries were ≥ 77.07%, and the precision values were ≤ 6.63%. The limit of detection and limit of quantification values were 1.05–1.32 and 3.50–4.20 μg/kg, respectively. The method was successfully applied for the analysis of piperazine in egg (whole egg, albumin and yolk) samples.