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

A competitive direct enzyme-linked immunosorbent assay (CD-ELISA) for hista mine 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 limi ts and mean recoveries (10-1000 mg/kg) were 2 mg/kg and 93% for CD-ELISA an d 1 mg/kg and 99% for RP-HPLC, respectively. Analysis of 50 commercial chee ses according to both methods showed good agreement for histamine (r = 0.97 9; concentration range = 2-1800 mg/kg). At a threshold level of 10 mg/kg, t he ELISA gave no false-negative and three false-positive results. The resul ts show that the ELISA is suitable for the determination of histamine in ch eese.