Development of a selective clean-up method using immobilized antibody and its application to HPLC and GC/MS determination of a carbacyclin derivative, CS-570, in plasma.
Methods for the determination of CS-570, a chemically stable prostacyclin analogue, in plasma were developed by using an immobilized antibody column followed by fluorescence HPLC and GC/MS. The CS-570 antibody, obtained from rabbit plasma by giving CS-570-BSA for a few months, was coupled to Sepharose 4B and used as extraction phase for sample clean-up and extraction of the drug. A plasma sample was applied to this column, washed with water and the drug was eluted with 90% acetonitrile. 0.02% (w/v) 9-anthryldiazomethane (ADAM) was added to the extract to form a fluorescent derivative. The CS-570-ADAM adduct exhibited high sensitivity when applied to HPLC with fluorescence detection and column switching. The detection limit was 1 ng/mL when 1 mL of plasma was available. Additionally, a pentafluorobenzyltrimethylsilyl derivative of CS-570 showed excellent sensitivity when determined by capillary GC interfaced to negative ion chemical ionization MS using a stable isotope labelled analogue as an internal standard.
Nakagawa
A
A
Analytical and Metabolic Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan.
Matsushita
Y
Y
Muramatsu
S
S
Tanishima
Y
Y
Hirota
T
T
Takasaki
W
W
Kawahara
Y
Y
Takahagi
H
H
D000818:Animals; D000906:Antibodies; D002851:Chromatography, High Pressure Liquid; D011464:Epoprostenol / Q000097:blood* / Q000276:immunology / Q000031:analogs & derivatives; D008401:Gas Chromatography-Mass Spectrometry; D051381:Rats
D000906:Antibodies; C038991:R 59274; D011464:Epoprostenol