Determination of histamine in the whole blood of colon cancer patients.

The aim of the present work is to investigate whether histamine assay could be useful in detecting the presence of primary cancer. The high-performance liquid chromatographic (HPLC)-based o-phthalaldialdehyde (OPA) histamine derivatization assay was investigated with respect to several variables, dramatization reagent concentration, organic solvent requirement, derivatization time and counter-ion effect on chromatographic separation. The OPA histamine assay, in the absence of added -SH groups, was found to detect histamine in whole blood samples with relative standard deviations <14% and recoveries not less than 90%. The assay showed high selectivity towards other aminic-containing compounds and a detection limit of 18 nM of histamine was evaluated. Calibration curves in the range 50-500 nM were obtained by using histamine standards in 0.1 M HCl with a regression coefficient value (r(2)) of 0.9969. In order to assess the usefulness of this assay in primary tumor monitoring, two groups of individuals, 29 controls and 29 colon cancer patients were selected, and serum levels of histamine, carcinogen embrionary antigen (CEA), carcinogen antigen 19.9 (CA19.9), and tumor staging, were determined. A significant histamine reduction (P=0.028) between controls (180.12+/-70.4 nM) and patients (134.5+/-90.3 nM) was found, and a cut-off value of 157.5 nM was extrapolated as intercept point of sensitivity and specificity curves. Fifty percent of patients showed a histamine value below the cut-off, while 45.8 and 8.3% of patients were positive for CEA and CA19.9, respectively. No correlation was found between Tumor Node Metastasis staging and histamine amount, indicating that this marker is not related to the tumor mass. Our data suggest that histamine level, together with other classical tumor markers, could be a potentially interesting tumor marker in colon cancer monitoring.