Micellar Liquid Chromatography Determination of Spermine in Fish Sauce after Derivatization with 3,5-Dinitrobenzoyl Chloride

A practical liquid chromatographic method has been developed for the selective determination of the levels of spermine in anchovy sauce after derivatization with 3,5 dinitrobenzoyl chloride. The micellar liquid chromatographic separation proposed here uses a C18 column ( 125x4.6 mm), followed by detection of spermine derivative at 260 nm. Elution of the analyte was performed using a mobile phase of 0.15 M SDS-4% (v/v) 1-pentanol-pH 7 running under isocratic mode at 25°C. Validation parameters were linearity (2-100 μg/mL,r2>0.999), detection and quantification limits (0.4 and 1.2 μg/mL, resp.), precision (less than 3.6%), accuracy (93.3-101.1%), and robustness (less than 4.8%). These results are in agreement with the requirements of the FDA guidelines. The proposed method was successfully applied to the monitorization of spermine formation in unsalted and salted fish sauce samples. The suggested methodology was found useful in routine analysis of spermine in fish sauce samples