An automated derivatization with dabsyl chloride in combination with a high-performance liquid chromatographic analysis is described for the determination of biogenic amines in complex matrices. The sample clean-up procedure consists of an ultrafiltration step, resulting in average recoveries for cheese in the range of 88% to 100%. A linear relation between the area of the peak and amine concentration was observed between 0.5 and 500 pmol for all amines under investigation and the detection limits ranged between 0.34 and 0.76 pmol. The average repeatability of both the performance of the chromatographic determination and the whole method, including sample preparation, examined using cheese samples containing different amounts of biogenic amines was found to be between 2.0% and 3.7%. The method was applied to the analysis of several types of food and feed, selected examples are given by the separation of dabsyl derivates from cheese, wine, and salami-type sausage.
Summary -An efficient method for the simultaneous determination of free amino acids (FAA) and biogenic amines (BA) was applied to the analysis of presumed raw milk cheeses of the following types:Emmental (/1 = 18), Bergkâse (/1 = 28), semi-hard cheeses (/1 = 18) and soft cheeses (11 = 41). The absolute amounts of FAA and BA largely varied, but the relative amounts could be classified into four groups independently of cheese type and ripening: i) glutamic acid, lysine, leucine (10-20% of total FAA); ii) proline, phenylalanine, valine (6-10%); iii) methionine, alanine, glutamine, isoleucine (3-6%); iv) BA and residual FAA « 3%). As the formation of BA is strictly dependent on the decarboxylase activity of microorganisms -raw milk flora, starter, ripening and contaminating bacteria -a large variation within cheese types and even individual cheeses occurred. The use of bactofugated milk for the production of Emmental cheese resulted in a decrease of putrescine and cadaverine, but did not significantly influence the formation of histamine and tyramine. When comparing cheese samples of the same type but from different heat treatments of cheese milk or curd, FAA and BA metabolites showed various tendencies; in particular, BA were not generally reduced upon higher heat treatment. Furthermore, hard cheese samples with strong sensory defects showed considerable amounts of cadaverine (up to 1800 mg/kg) as weil as ô-amino valerie acid (up to 2200 mg/kg), which in regular cheese samples of the same type and age was found to be remarkably lower (100-200 mglkg). The combined data for FAA and BA provide useful information on chee se ripening and quality control.Cree amino acid / biogenic amine / cheese ripening Oral communication at the IDF Symposium 'Ripening and Quality of Chee se s' , Besançon, France, February 26-28, 1996. 102 1 Krause et al Résumé -Caractérisation de la maturation du fromage par ses acides aminés libres et ses amines biogènes, et influence de la bactofugation et du traitement thermique du lait. Une méthode performante a été utilisée pour l'analyse simultanée des acides aminés libres (AAL) et des amines biogènes (AB) de fromages supposés être fabriqués au lait cru. de types emmental (Il = 18), de montagne (Il = 28), à pâtes mi-dure (Il = 18) et molle (Il = 41). La teneur absolue totale en AAL et en AB variait beaucoup, mais a pu néanmoins être distribuée en quatre classes distinctes, indépen-dantes du type de fromage considéré et du degré d'affinage, soit: i) l'acide glutamique, la lysine et la leucine (10-20 % du total); ii) la proline, la phénylalanine et la valine (6-10 %); iii) la méthio-nine, l'alanine, la glutamine et l'isoleucine (3-6 %) ; iv) les AB et les autres acides aminés « 3 %).Comme la formation des AB ne dépend que de l'activité de la décarboxylase des micro-organismes, de la flore originelle du lait, des levains, des bactéries d'affinage et des contaminants, on a observé une grande variation de concentration de ces constituants, tant entre les différentes sortes qu'à l'intérieur d'un mê...
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