Analytical Method to Evaluate Gizzerosine in Fishmeal After Diazonium Derivatization Using High-Performance Liquid Chromatography

A high-performance liquid chromatography method for the analysis of gizzerosine in fishmeal was developed. The method was performed using an ODS column (250 × 4.6 mm) and achieved a gizzerosine total analysis time of approximately 35 min. Gizzerosine was extracted from fishmeal with a 0.1-N HCl solution, diazotized with sulfanilic acid (Pauly’s reagent), and successfully separated from other reagent-positive components in fishmeal extracts. Using this method, the calibration curve of standard gizzerosine was estimated to be within the range of 1 to 250 mg/l (r² = 0.9942). The coefficient of variation (relative standard deviation) of gizzerosine derivatization was 0.27%. The mean recovery was 96.2% with addition of 0.1, 1, 5, 10, and 15 μg/100 g of gizzerosine in fishmeal. Moreover, the limit of detection for spiked feed blanks, based on a S/N of 3, was 0.61 mg/kg. This method was also efficient in detection of the wavelength of the analyte derivatives as well as in clean sample preparation. Therefore, this method could be used to analyze gizzerosine in fishmeal.