Abstract
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 (125×4.6 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.
Highlights
Biogenic amines are biological metabolites present in foods either as natural products or after fermentation, decay microbial contamination, decomposition, or putrefaction processes [1]
Mobile phases were aqueous solutions of Sodium dodecyl sulphate (SDS) buffered at pH 7 with phosphate buffer and the stationary phase was a C18 column [2, 6]
The salted sample shows no microbacterial contamination and can be eaten without risk. These results indicate that salting is an efficient method to prevent microbial contamination during anchovy sauce storage
Summary
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. 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 [1,2,3]. Biogenic amines can be found in semen and some microalgae, together with related molecules like spermine and spermidine. The determination of spermine is of the utmost importance to assure that the fish sauce can be eaten without health risk [4, 5]
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