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KMID : 1101420150470040306
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Korean Journal of Clinical Laboratory Science
2015 Volume.47 No. 4 p.306 ~ p.312
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Comparison of Methods for Measuring Histamine by ELISA and HPLC-MS Assay In Vitro
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Lee In-Hee
Kim Yoo-Hyun
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Abstract
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The measurement of histamine is to determine the degree of allergy because the allergic reaction can lead to the release of histamine. In general, the antigen-antibody reaction was quantified by measuring absorbance using a microplate reader. In this study, we compare the method using a general antigen-antibody reaction and the method using a high performance liquid chromatography mass spectrometer (HPLC-MS) of chemical analysis in the measurement of histamine secretion. The cell line used was RBL-2H3, an allergic reaction was induced by stimulation with C48/80 (compound 48/80). Allergy-induced cells degranulation rate was confirmed by measurement of β-hexosaminidase and cytotoxicity was performed for the validity of the experiment. The quantitative determination of histamine showed a significant difference, since the quantitative limit of the measurement by the antigen-antibody reaction was 10.257 ppm while the quantitative limit of the measurement by HPLC-MS was 0.020 ppm. Measurement of histamine in allergic activity and anti-allergy tests showed that the HPLC-MS analysis rather than the analysis of the antigen-antibody reaction is a more precise and accurate test.
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KEYWORD
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Allergy, β-hexosaminidase, Histamine, High performance liquid chromatography mass spectrometer
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