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 ( ？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, ), 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. 1. Introduction Biogenic amines are biological metabolites present in foods either as natural products or after fermentation, decay microbial contamination, decomposition, or putrefaction processes [1]. They are largely responsible for the foul odour of putrefying flesh, as well as contributing to the odour of such processes as bad breath and bacterial vaginosis. Biogenic amines are also involved in local immune responses, neurotransmission, and chemotaxis of white blood cells. The consumption of an excess of biogenic amines, known as histaminic intoxication, is mainly related to heart, gastrointestinal, and skin diseases, as well as headache [1–3]. Food containing considerable amounts of these amines include alcoholic beverages, beef, chocolate, cheeses, fish, pork, and poultry. Biogenic amines can also be found in semen and some microalgae, together with related molecules like spermine and spermidine. In fact, spermine is formed from spermidine and can be found in a wide variety of organisms and tissues, as it is an essential growth factor in some bacteria. Thus, its detection and quantification is useful to assess the degree of bacterial contamination, mainly caused by incorrect handling or stocking conditions (freezer at ？18°C), in fish flesh, or derivates as fish sauce. Then the determination of spermine is of the utmost importance to assure that the fish sauce can be eaten without health risk [4, 5]. Several analytical methods have been developed for the determination of spermine. Among them, HPLC with UV-visible absorbance detection using 3,5-dinitrobenzoyl chloride (DNBZ-Cl) as a
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