High-performance liquid chromatographic determination of plasma histamine after pre-column derivatisation with o-phthaldialdehyde |
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Authors: | R Czerwonka D Tsikas G Brunner |
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Institution: | (1) Department of Gastroenterology and Hepatology, College of Medicine Hannover, Oststadt Hospital, Podbielskistr. 380, D-3000 Hannover 51, FRG |
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Abstract: | Summary A reversed-phase high-performance liquid chromatographic (HPLC) method has been developed for the sensitive and highly selective
determination of histamine in plasma. This method includes selective extraction of histamine from plasma, pre-column derivatisation
in aqueous phase with o-phthaldialdehyde (OPA) and HPLC analysis. The fluorescence of the histamine-OPA-complex was monitored
at wavelengths of 350nm excitation and 450nm emission, after isocratic eluation with a mixture of 0.2 M NaCl and methanol.
The reproducibility of this method including extraction, derivatisation and detection of histamine was >95% in a range of
0.35–17.6 pmol. The HPLC precision was 99±1% at 4 pmol of histamine. The lower limit of detection was 88fmol. A significantly
increased concentration of plasma histamine was detected in patients (n=46) with various liver diseases (0.3–5.2 ng/ml). In
comparison the plasma histamine levels of healthy blood donors were in the range of 0.0–0.4 ng/ml (p<0.01). |
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Keywords: | High-performance liquid chromatography Precolumn derivatisation o-Phthaldialdehyde Plasma histamine determination Liver disease |
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