Development and validation of an HPLC-FLD method for rapid determination of histamine in skipjack tuna fish (Katsuwonus pelamis)

This study compared and validated two methods of high-performance liquid chromatography: fluorescence detection (HPLC-FLD) and UV–Vis detection. Recoveries greater than 55% at all spiking levels using UV–Vis detection could not be obtained. However, derivatization with pre-column o-phthalaldehyde (OPA) was found to efficiently enhance the method sensitivity. The analytical method was validated based on the following criteria: linearity, sensitivity, accuracy, precision, repeatability, and recovery. For fluorescence detection, excellent linear correlation with R2 = 0.9977 was observed over the range of 5.0 to 100.0 mg/L. The minimum detection limit was calculated to be 1.5 mg/kg, while the limit of quantification (LOQ) value of the validated method was determined to be 4.5 mg/kg. Good recoveries (RSD% = 1.35) were obtained at all spiking levels ranging between 0 and 200 mg/kg. The proposed method was found to be suitable, selective, and rapid for the determination of histamine.