Sensitive determination of salmon calcitonin, by means of pre-column derivatization, HPLC and fluorometric determination

A high sensitive analytical method for salmon calcitonin (sCT) was established by means of derivatization with a fluorescence labelling agent, DBD-F, followed by HPLC separation and fluorescence detection at 558 nm with excitation at 430 nm. Under optimized conditions (DBD-F, 0.1 M borate buffer (pH...

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Bibliographic Details
Published inBiomedical chromatography Vol. 9; no. 1; p. 52
Main Authors Fukuda, T, Ishikawa, K, Imai, K
Format Journal Article
LanguageEnglish
Published England 01.01.1995
Subjects
Analgesics - analysis
Calcitonin - analysis
Calibration
Chromatography, High Pressure Liquid
Fluorescent Dyes
Hydrogen-Ion Concentration
Mass Spectrometry
Oxazoles
Sodium Dodecyl Sulfate
Spectrometry, Fluorescence
Sulfhydryl Reagents
Sulfonamides
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Summary:A high sensitive analytical method for salmon calcitonin (sCT) was established by means of derivatization with a fluorescence labelling agent, DBD-F, followed by HPLC separation and fluorescence detection at 558 nm with excitation at 430 nm. Under optimized conditions (DBD-F, 0.1 M borate buffer (pH 8.5):acetonitrile (70:30, v/v)), three molecules of DBD-F reacted with one sCT molecule in the presence of 1 mM sodium dodecyl sulphate (SDS) at 50 degrees C for 3 h. The detection limit for the sCT derivative was 20 fmol. A linear relationship was obtained between the amount of sCT on column and the peak heights in the range of 35-8000 fmol.
ISSN:0269-3879
DOI:10.1002/bmc.1130090111