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Comparison of ELISA and HPLC for the determination of histamine in cheese

Aygun, O., Schneider, E., Scheuer, R., Usleber, E., Gareis, M., Martlbauer, E.
Journal of agricultural and food chemistry 1999 v.47 no.5 pp. 1961-1964
cheeses, histamine, tyramine, putrescine, cadaverine, liquid chromatography, high performance liquid chromatography, reversed-phase high performance liquid chromatography, enzyme-linked immunosorbent assay, soft cheeses, semisoft cheeses
A competitive direct enzyme-linked immunosorbent assay (CD-ELISA) for histamine in cheese was compared with a reversed-phase liquid chromatography (RP-HPLC) method. Cheese was homogenized with phosphate-buffered saline (PBS), centrifuged, and filtered, and the supernatant was diluted with PBS for CD-ELISA. For RP-HPLC, biogenic amines (histamine, tyramine, putrescine, and cadaverine) were derivatized with 9-fluorenylmethylchloroformate, followed by reversed-phase chromatography and fluorescence detection. Detection limits and mean recoveries (10-1000 mg/kg) were 2 mg/kg and 93% for CD-ELISA and 1 mg/kg and 99% for RP-HPLC, respectively. Analysis of 50 commercial cheeses according to both methods showed good agreement for histamine (r = 0.979; concentration range 2-1800 mg/kg). At a threshold level of 10 mg/kg, the ELISA gave no false-negative and three false-positive results. The results show that the ELISA is suitable for the determination of histamine in cheese.