Bioanalysis and preliminary pharmacokinetics of the acridonecarboxamide derivative GF120918 in plasma of mice and humans by ion-pairing reversed-phase high-performance liquid chromatography with fluorescence detection

• Published in: Journal of chromatography. B, Biomedical sciences and applications Vol. 759; no. 1; pp. 135 - 143
• Main Authors: Kemper, E.M., Jansen, B., Brouwer, K.R., Schellens, J.H.M., Beijnen, J.H., van Tellingen, O.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 05.08.2001; Elsevier Science
• Subjects: Acridines - pharmacokinetics; Acridonecarboxamide; Analysis; Animals; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; General pharmacology; GF120918; Humans; Isoquinolines - blood; Isoquinolines - pharmacokinetics; Medical sciences; Mice; Pharmacology. Drug treatments; Reproducibility of Results; Sensitivity and Specificity; Spectrometry, Fluorescence; Tetrahydroisoquinolines; GF120918; Acridonecarboxamide; Human; Validation; Protein inhibitor; Biological fluid; Fluorescence detector; Rodentia; P Glycoprotein; Ion pair; Oral administration; HPLC chromatography; Blood plasma; Vertebrata; Reversed phase chromatography; Mammalia; Mouse; Animal; Carboxamide; Pharmacokinetics; Quantitative analysis
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(01)00207-9

We have developed and validated a sensitive and selective method for the determination of the P-glycoprotein modulator GF120918 in murine and human plasma. Chlorpromazine is used as internal standard. Sample pretreatment involves liquid–liquid extraction with tert-butyl methyl ether. Chromatographic separation is achieved by reversed-phase high-performance liquid chromatography using a Symmetry C 18 column and detection was accomplished with a fluorescence detector set at excitation and emission wavelengths of 260 and 460 nm, respectively. The mobile phase consists of acetonitrile–50 m M ammonium acetate buffer, pH 4.2 (35:65, v/v). To achieve good separation from endogenous compounds and to improve the peak shape the counter-ion 1-octane sulfonic acid (final concentration 0.005 M) was added to the mobile phase. The lower limit of quantitation was 5.7 ng/ml using 200 μl of human plasma and 23 ng/ml using 50 μl of murine plasma. Within the dynamic range of the calibration curve (5.7–571 ng/ml) the accuracy was close to 100% and within-day and between-day precision were within the generally accepted 15% range. The stability of GF120918 was tested in plasma and blood from mice and humans incubated at 4°C, room temperature, and 37°C for up to 4 h. No losses were observed under these conditions. This method was applied to study the pharmacokinetics of orally administered GF120918 in humans and mice. The sensitivity of the assay was sufficient to determine the concentration in plasma samples obtained up to 24 h after drug administration.