Determination of the Triacylglycerol Fraction in Fish Oil by Ultra Performance and Comprehensive Liquid Chromatography Techniques with the Support of Gas Chromatography and Mass Spectrometry Data 
Marco Beccaria1, Rosaria Costa1, Marianna Oteri1, Francesco Cacciola2, Giorgia Purcaro3, Paola Dugo1,3,4, Luigi Mondello1,3,4 1“Scienze del Farmaco e Prodotti per la Salute” Department, University of Messina, viale Annunziata, 98168 – Messina, Italy
2"Scienze dell'Ambiente, della Sicurezza, del Territorio, degli Alimenti e della Salute" Department, University of Messina, Viale F. Stagno d'Alcontres 31, 98166 Messina, Italy  Department, University of Messina, viale Annunziata, 98168 – Messina, Italy
3University Campus Bio-Medico of Rome, Via Álvaro del Portillo, 21 - 00128 Rome, Italy 4 Chromaleont s.r.l., c/o “Scienze del Farmaco e Prodotti per la Salute” Department, University of Messina, viale Annunziata, 98168 – Messina, Italy
 
Fish oil made from menhaden (Brevoortia Tyrannus) can be used as a dietary supplement for the presence of high levels of the long-chained omega-3 fatty acids, viz. eicosapentanoic and docosahexanoic. Studies on ingestion of marine polyunsaturated fatty acids, in particular of omega-3 series, in patients with rheumatoid arthritis and other diseases characterized by chronic inflammation, highlighted the benefits on such lipid dysfunctions. 
In this work, different analytical approaches were developed and compared, in terms of peak capacity, for characterization of triacylglycerol fraction in fish oil. In particular, i) four columns were serially coupled, for a total length of 60 cm of shell-packed stationary phase, and operated under ultra high pressure conditions; ii) comprehensive LC in both stop-flow and off-line modes, using silver ion chromatography with a silver-ion column and non-aqueous reverse-phase liquid chromatography with a C18 column were tested. The use of mass spectra attained by atmospheric pressure chemical ionization for all LC approaches, and the fatty acids methyl esters profile of menhaden oil obtained by gas chromatography analysis, greatly supported the elucidation of the triacylglycerol content in menhaden oil.