Comparison of two enzymatic immunoassays, high resolution mass spectrometry method and radioimmunoassay for the quantification of human plasma histamine

• LC–HRMS and Immunotech® EIA both displayed high correlation with RIA.
• LC–HRMS displayed high sensitivity and specificity.
• 7.88 nM and 14.39 nM was determined as new cut-offs for LC–HRMS and Immunotech® EIA.

Histamine (HA) is one of the main immediate mediators involved in allergic reactions. HA plasma concentration is well correlated with the severity of vascular and respiratory signs of anaphylaxis. Consequently, plasma quantification of HA is useful to comfort the diagnosis of anaphylaxis. Currently, radioimmunoassay (RIA) is the gold standard method to quantify HA due to its high sensitivity, but it is time consuming, implicates specific formations and cautions for technicians, and produces hazardous radioactive wastes. The aim of this study was to compare two enzymatic immunoassays (EIA) and one in-house liquid chromatography high-resolution mass spectrometry method (LC–HRMS) with the gold standard method for HA quantification in plasma samples of patients suspected of anaphylaxis reactions. Ninety-two plasma samples were tested with the 4 methods (RIA, 2 EIA and LC–HRMS) for HA quantification. Fifty-eight samples displayed HA concentrations above the positive cut-off of 10 nM evaluated by RIA, including 18 highly positive samples (>100 nM). This study shows that Immunotech® EIA and LC–HRMS concentrations were highly correlated with RIA values, in particular for samples with a HA concentration around the positive cut-off. In our hands, plasma concentrations obtained with the Demeditec Diagnostics® EIA correlated less with results obtained by RIA, and an underestimation of plasma HA levels led to a lack of sensitivity. In conclusion, this study demonstrates that Immunotech® EIA and LC–HRMS method could be used instead of RIA to assess plasma HA in human diagnostic use.

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