Wine Chemistry and Biochemistry

(Steven Felgate) #1

180 M.V. Moreno-Arribas and M.C. Polo


concentrations. Several techniques have been developed for biogenic amine deter-


mination in foods, some to analyse one specific biogenic amine (histamine or tyra-


mine), and others to detect more than one simultaneously. Initially, spectrofluorimet-


ric techniques were used to determine biogenic amines (Vidal-Carou et al. 1990b).


More recently, chromatographic methods are the techniques most commonly applied


to analyse several biogenic amines simultaneously.


On the other hand, qualitative measurements have also been described to indi-


cate the presence of amines in wine. The potential of biogenic amines to appear


in wine can be determined by using molecular tools which detect the presence of


decarboxylase-positive microorganisms.


6A.3.4.1 Quantification by Liquid Chromatography


Of the wide range of analytical techniques to detect and quantify biogenic amines


in wines described in the literature, reverse phase high-power liquid chromatogra-


phy (RP-HPLC) is the one most widely used in laboratories, since it can obtain the


most information about all the amines identified in must and wine (Lehtonen 1996;


Romero et al. 2002; Torrea and Anc ́ın 2002; V ́azquez-Lasa et al. 1998; Marcobal


et al. 2005b; Hern ́andez-Orte et al. 2006b; G ́omez-Alonso et al. 2007). In fact, one


RP-HPLC technique is currently being adopted as a reference technique by the Inter-


national Organization of Vine and Wine (OIV).


Owing to the absence of chromophores from these compounds, it is necessary


to form derivatives that are absorbed in ultraviolet or are fluorescent to facilitate


detection. For this purpose, dansyl-chloride andO-phthaldialdehyde (OPA) are
the reagents most used. In the case of dansyl-chloride, the reaction is carried out


before chromatographic separation, while with OPA it can be done before the col-


umn or after (V ́azquez-Lasa et al. 1998; Marcobal et al. 2005b). Other derivatiz-


ing reagents used, of more general application, are 9-fluorenylmethylchloroformate


(FMOC), aminoquinolyl-N-hydroxysuccinimidyl caramate (AQC) (Hern ́andez-Orte


et al. 2006b), and diethyl ethoxymethylenemalonate (DEEMM) (G ́omez-Alonso


et al. 2007). Of all these derivatives, the most used is OPA, because of its rapid


one-step derivatization reaction and the possibility of automating the reaction, which


increases the reproducibility of the analytical method, among other reasons.


Besides HPLC methods, over the past few years other chromatographic tech-


niques applicable to biogenic amine analysis have been performed. Among oth-


ers, micellar liquid chromatography (MLC) (Gil-Agust ́ı et al. 2007); micellar


electrokinetic chromatography (MECC) separation with laser-induced fluorescence


(LIF) detection (Nouadje et al. 1997), nanoliquid chromatography with UV detec-


tion (Hern ́andez-Borges et al. 2007), ion-pair liquid chromatography (Hlabangana


et al. 2006), and liquid chromatography-electronspray ionisation ion trap mass spec-


trometry (Mill ́an et al. 2007) are used.


6A.3.4.2 Quantification by Capillary Electrophoretic Techniques


The attraction of using capillary electrophoresis (CE) methods mainly lies in the


short analysis time, high separation efficiencies, and reduced consumption of

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