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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.