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 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.
Fukuda
T
T
Tsumura & Co., Ibaraki, Japan.
Ishikawa
K
K
Imai
K
K
D000700:Analgesics / Q000032:analysis*; D002116:Calcitonin / Q000032:analysis*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry; D010080:Oxazoles; D012967:Sodium Dodecyl Sulfate; D013050:Spectrometry, Fluorescence; D013439:Sulfhydryl Reagents; D013449:Sulfonamides
D000700:Analgesics; D005456:Fluorescent Dyes; D010080:Oxazoles; D013439:Sulfhydryl Reagents; D013449:Sulfonamides; D012967:Sodium Dodecyl Sulfate; C028815:salmon calcitonin; D002116:Calcitonin; C060548:4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole