Kl. Napoli et Bd. Kahan, SAMPLE CLEANUP AND HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC TECHNIQUESFOR MEASUREMENT OF WHOLE-BLOOD RAPAMYCIN CONCENTRATIONS, Journal of chromatography B. Biomedical applications, 654(1), 1994, pp. 111-120
Citations number
10
Categorie Soggetti
Chemistry Analytical
Journal title
Journal of chromatography B. Biomedical applications
Determination of rapamycin (RAPA), a potential adjunct to cyclosporin
immunosuppressive therapy in solid organ transplantation, presents a c
hallenge because of the agent's unusual chemical properties. RAPA is n
ot readily extracted from biological matrices, is light-sensitive, and
exists in numerous isomeric forms. Using liquid-liquid extraction tec
hniques with tert.-butyl methyl ether followed by ethanol, one achieve
s 96% recovery of RAPA and 70% recovery of the internal standard, beta
-estradiol-3-methyl ether, from human whole blood. Subsequent high-per
formance liquid chromatography (HPLC) utilizing two heated reversed-ph
ase C18 columns in tandem and an 85% methanol-water mobile phase with
ultraviolet detection at 276 nm provides measurement of RAPA from 1-ml
samples of human whole blood. The chromatography requires less than 4
0 min per sample. The assay exhibits less-than-or-equal-to 10% standar
d error of the mean and less-than-or-equal-to 12% coefficient of varia
tion over the concentration range 2-100 ng/ml. The method has been tes
ted using pharmacokinetic profiles from renal transplant recipients re
ceiving bolus intravenous RAPA infusions.