Comparison of ELISA and HPLC for the determination of histamine in cheese.
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.
Aygün
O
O
Institute for Toxicoloy and Microbiology, Federal Center for Meat Research, Kulmbach, Germany. Vete.
Schneider
E
E
Scheuer
R
R
Usleber
E
E
Gareis
M
M
Märtlbauer
E
E
D001679:Biogenic Amines / Q000032:analysis; D002611:Cheese / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006632:Histamine / Q000032:analysis*; D012680:Sensitivity and Specificity
D001679:Biogenic Amines; D006632:Histamine