Talanta 85 (2011) 363–369 Contents lists available at ScienceDirect Talanta journal homepage: www.elsevier.com/locate/talanta A new method for the determination of biogenic amines in cheese by LC with evaporative light scattering detector Donatella Restuccia ∗ , U. Gianfranco Spizzirri, Francesco Puoci, Giuseppe Cirillo, Manuela Curcio, Ortensia I. Parisi, Francesca Iemma, Nevio Picci Dipartimento di Scienze Farmaceutiche, Università della Calabria, Edificio Polifunzionale, Arcavacata di Rende (CS) 87036, Italy article info Article history: Received 7 December 2010 Received in revised form 15 March 2011 Accepted 28 March 2011 Available online 5 April 2011 Keywords: Biogenic amines Evaporative light scattering detector Cheese Food safety Food quality abstract This paper presents a new LC method with evaporative light scattering detection (ELSD), for the separation and determination of the biogenic amines (histamine, spermidine, spermine, tyramine, putrescine and -phenylethylamine) which are commonly present in cheese, as their presence and relative amounts give useful information about freshness, level of maturing, quality of storage and cheese authentication. The LC–ELSD method is validated by comparison of the results with those obtained through LC–UV deter- mination, based on a pre-column dansyl chloride derivatisation step. The obtained data demonstrate that both methods can be interchangeably used for biogenic amines determination in cheese. The new LC–ELSD method shows good precision and permits to achieve, for standard solutions, limit of detec- tion (LOD) values ranging from 1.4 to 3.6 mg L −1 and limit of quantitation (LOQ) values ranging from 3.6 to 9.3 mg L −1 . The whole methodology, comprehensive of the homogenization–extraction process and LC–ELSD analysis, has been applied in the analysis of a typical Calabria (Southern Italy) POD cheese, known as Caciocavallo Silano. The most aboundant amine found was histamine, followed, in decreasing order, by tyramine, spermine, putrescine, -phenylethylamine and spermidine, for a total amount of 127 mg kg −1 . This value does not represent a possible risk for consumer health, according to the toxicity levels reported in literature and regarded as acceptable. © 2011 Elsevier B.V. All rights reserved. 1. Introduction Bioactive amines are nitrogenous compounds that can be clas- sified by their formation pathway and biological functions. During the polyamine biosynthesis, they are formed by the activity of endogenous decarboxylase enzymes, naturally occurring in food, and play a variety of biological roles [1,2]; on the contrary, biogenic amines (BAs) are detrimental to health and originate in foods from decarboxylation of the corresponding amino acid and transam- ination of aldehydes and ketones by the activity of exogenous enzymes released by various microorganisms [3–6]. Therefore, BAs may be of endogenous origin at low concentrations even in non- fermented foods such as fruits, vegetables, meat, milk and fish, while high concentrations of these compounds can be found as a consequence of microbial activity in foods such as wine, fer- mented meat and fish products, cheese and fermented vegetables [2–4]. Since the consumption of food containing high concentra- tions of BAs may cause toxic reactions in susceptible individuals [2,3,6,7], in recent years considerable research has been under- ∗ Corresponding author. Tel.: +39 0984 493296; fax: +39 0984493163. E-mail address: donatella.restuccia@unical.it (D. Restuccia). taken to evaluate the presence of these compounds in various fermented, seasoned or conserved foodstuffs. Among foods com- monly associated with the presence of BAs, particular attention has been devoted to cheese. Many factors have been found to affect the production of biogenic amines in cheeses, including the presence of microorganisms decarboxylating free amino acids [8–12], the presence of spoiling microorganisms [13], the syner- gistic effects between microorganisms and the availability of free amino acids produced as an outcome of proteolysis levels [14–17]. Several extrinsic processing factors may also play an important role, namely, pH, salt-in-moisture levels and ripening temperature [18,19]. Because the content of BAs is influenced by ripening and quality of storage conditions, interest in the presence and amount of BA in food is always increasing, not only because of the potential toxicity but also because the content of BAs is an useful param- eter to evaluate the extent of ripening and the quality of storage [11,20,21]. Different analytical methods have been used to determine BAs in foods. Since most amines show neither natural UV absorption nor fluorescence, most methods require that amines should be deriva- tized before detection. Different chemical regents have been used for the amine analysis, such as dansyl chloride [22–27], dabsyl chlo- ride [28–30], o-phthlaldehyde [31–34], fluorescein isothiocyanate 0039-9140/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.talanta.2011.03.080