Quantification of riboflavin and niacin in anchovies (Engraulis enrasicolus) by HPLC-MS/MS triple quadrupole.

Introduction: Vitamins are crucial for maintaining good health in humans [1]. In foodstuffs, the vitamins B2 (riboflavin) and B3 (niacin) may be present in free and binding forms. Anchovies are a good source of those two vitamins, thus, the aim of this paper was to evaluate the content of vitamin riboflavin and niacin in anchovies samples provided by a company called “Dimar Sapore di Mare”. Methods: Different extraction procedures were tested (acidic/enzymatic hydrolysis and combination of them) and the best one was chosen for the accurate quantification of riboflavin, nicotinic acid and nicotinamide in different types of anchovy samples. UHPLC–MS/MS studies were performed using an HPLC-MS/MS triple quadrupole equipped with an ESI source operating in positive ionization mode. Results: The results obtained indicated that acidic hydrolysis alone was the best of the four optimized extraction procedures in terms of recoveries (range 96.2 - 105.3%, with RSDs lower than 9.2%), quantitative results obtained, and rapidity, and thus it was chosen for the analysis of vitamins B2 and B3 in 23 anchovy samples (both normal and canned ones). The application to real samples indicates that the highest niacin levels were found in normal anchovies (101.21-150.92 mg kg-1), while the highest level of riboflavin was in anchovies canned in olive oil (ranging from 2.90 to 3.35 mg kg-1). Conclusions: A new analytical method that uses HPLC-MS/MS triple quadrupole was developed for the first time for the analysis of riboflavin, nicotinamide and nicotinic acid in anchovies. Samples were extracted using different extraction procedures, and acidic hydrolysis (method 1) was found to be the best in terms of recoveries, quantitative results and rapidity. In order to understand the dietary intake of vitamins from different types of anchovies, the method was applied to 23 samples including eleven normal anchovy samples and twelve canned ones. References 1. A. Zafra-Gomez, A. Garballo, J.C. Morales, L.E. Garcia-Ayuso. Journal of Agricultural and Food Chemistry, 54 (2006), pp. 4531-4536. 2. A. Gentili, F. Caretti, G. D’Ascenzo, S. Marchese, D. Perret, D. Di Corcia, L.M. Rocca. Rapid Communications in Mass Spectrometry, 22 (2008), pp. 2029-2043.