Evaluation of the stable reaction products of histidine with formaldehyde or with other carbonyl compounds in dairy products

Combined formaldehyde (FA) can be evaluated in cheese and other dairy products by determining spinacine (6-carboxy-1,2,3,4-tetrahydroimidazopyridine), an imino acid arising from the reaction of FA with the -amino group of histidine. Other carbonyl compounds usually produced by fermentations can react themselves with histidine, forming molecules which interfere with the determination of spinacine. A sensitive (minimum detectable amount, 5 fmol spinacine) and interferencefree HPLC method, with precolumn derivatization with o-phtalaldehyde (OPA) and 9-fluorenylmethylchloroformate (FMOC), to evaluate these histidine reaction-compounds in dairy products is described. The clean-up of the FMOC-derivatized sample has been performed for the first time with solid-phase extraction (SPE) on an amino cartridge. The method was applied to samples of casein and differently ripened cheese, the origins of which were either known or commercial, in order to quantify the natural level of spinacine (0.7–2.7 ppm) probably deriving from biogenic FA, and to detect whether FA was used in processing.