Determination of Semicarbazide in Fresh Egg and Whole Egg Powder by Liquid Chromatography Tandem Mass Spectrometry: Inter-laboratory Validation Study

An inter-laboratory validation study according to harmonized protocols, was carried out to evaluate the effectiveness of a liquid chromatography tandem mass spectrometry (LC/MS/MS) method for the determination of semicarbazide (SEM) in fresh whole egg and in a industrially processed whole egg powder. An aliquot of the sample was extracted with hydrochloric acid and derivatized with 2-nitrobenzaldehyde, using 1,2-[15N2, 13C] SEM as internal standard. The extract was neutralized and purified on a solid phase extraction (SPE) cartridge. SEM was determined by reverse phase LC with detection by MS/MS. Five fresh egg samples, three of them obtained from nitrofurazone incurred hens, one spiked with 50 µg•kg-1 SEM and a blank sample, and five industrial whole egg powder samples, three of them spiked with fresh whole egg from incurred hens, one sample spiked with 350 µg•kg-1 SEM and one blank sample, were sent to 15 laboratories from 10 different European countries from which results were obtained from 12 participants. Recoveries for the fresh egg and the egg powder were 105.3 % and 121.3%, respectively. The relative standard deviation for repeatability (RSDr) ranged from 2.9 % to 9.3% and the relative standard deviation for reproducibility (RSDR) ranged from 21.0% to 38.1 %. The method showed acceptable within-laboratory and between laboratory precision for all two matrices, as evidenced by HORRAT values, at the target levels of determination for SEM.JRC.D.8-Food safety and qualit