Reversed-phase high-performance liquid chromatography of salmon calcitonin and its degradation products in biological samples using column switching and flow-through radioisotope detection.
For the determination of salmon calcitonin and its degradation products in biological samples, a reversed-phase HPLC method with column switching and flow-through radioisotope detection has been developed using high specific activity [125I]salmon calcitonin. Effects of the precolumn packing material and washing solvent were examined in terms of [125I]salmon calcitonin recovery. Spiked samples of [125I]salmon calcitonin in plasma and kidney homogenate were injected onto a LiChroprep RP-8 precolumn after dilution with 0.1% trifluoroacetic acid. After washing the polar interfering compounds with 0.1% trifluoroacetic acid, the concentrated [125I]salmon calcitonin and its degradation products were eluted and separated on a W-Porex C18 column with a gradient of 0.1% trifluoroacetic acid in acetonitrile-water. Detection and calibration of [125I]salmon calcitonin were possible down to picogram levels. Reproducible kinetic data for the degradation of intact [125I]salmon calcitonin were possible down to picogram levels. Reproducible kinetic data for the degradation of intact [125I]salmon calcitonin by rat kidney homogenate could be traced.
Lee
H S
HS
Screening and Toxicology Center, Korea Research Institute of Chemical Technology, Yoosung, Taejeon, South Korea.
Lee
J S
JS
Lee
H
H
Jung
Y S
YS
DeLuca
P P
PP
Lee
K C
KC
D000818:Animals; D002116:Calcitonin / Q000032:analysis* / Q000097:blood / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D007668:Kidney / Q000737:chemistry*; D051381:Rats; D015203:Reproducibility of Results
C028815:salmon calcitonin; D002116:Calcitonin