Liquid-chromatographic measurement of cyclosporin A and its metabolites in blood, bile, and urine.
Using solid-phase extraction columns and "high-performance" liquid-chromatographic (HPLC) analysis, we could determine cyclosporin A and nine of its metabolites in blood, bile, and urine. To facilitate calculations of concentrations of cyclosporin A and its metabolites from the chromatograms, we used cyclosporin D as internal standard. For the HPLC analysis we used two sequential 250-mm analytical columns filled with reversed-phase octyl (C8) sorbent, eluting with a concave gradient of water, adjusted to pH 3.0 with phosphoric acid, and acetonitrile. Peaks were detected at 205 nm. For characterization of the chromatographic peaks, we isolated, by semi-preparative HPLC, 32 fractions representing peaks potentially related to cyclosporin A metabolites and re-injected them into the HPLC system under the same conditions as authentic cyclosporin A metabolites. Analytical recovery was 70-80%. The inter-assay CV for bile was 7.2%, for urine 12.3%. The method was used for routine monitoring of cyclosporin A and its metabolites.
Christians
U
U
Abteilung Allgemeine Pharmakologie, Medizinische Hochschule Hannover, Hannover F.R.G.
Zimmer
K O
KO
Wonigeit
K
K
Maurer
G
G
Sewing
K F
KF
D001646:Bile / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D003524:Cyclosporins / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D016030:Kidney Transplantation; D016031:Liver Transplantation; D013223:Statistics as Topic
D003524:Cyclosporins; C050026:cyclosporin D