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Sensitive determination of salmon calcitonin, by means of pre‐column derivatization, HPLC and fluorometric determination
Author(s) -
Fukuda Terumi,
Ishikawa Kazuyuki,
Imai Kazuhiro
Publication year - 1995
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130090111
Subject(s) - chemistry , derivatization , chromatography , detection limit , high performance liquid chromatography , salmon calcitonin , acetonitrile , fluorescence , derivative (finance) , sodium , linear range , fluorescence spectroscopy , analytical chemistry (journal) , calcitonin , organic chemistry , medicine , physics , quantum mechanics , financial economics , economics
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 o 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.