Quantitation of imidazo[1,2-a]quinoxaline derivatives in human and rat plasma using LC/ESI-MS

• Published in: Journal of separation science 2009-05, Vol.32 (9), p.1363-1373
• Main Authors: Khier, Sonia, Moarbess, Georges, Deleuze-Masquefa, Carine, Solassol, Isabelle, Margout, Delphine, Pinguet, Frédéric, Bonnet, Pierre-Antoine, Bressolle, Françoise M.M
• Format: Article
• Language: English
• Subjects: 2-a]quinoxaline; Analysis; Analytical chemistry; Animals; Biological and medical sciences; Chemical Sciences; Chromatography, Liquid; Drug Stability; General pharmacology; Human and rat matrices; Humans; Imidazo; Imidazo[1,2‐a]quinoxaline; Imidazoles - blood; Imidazoles - chemistry; Imidazoles - pharmacokinetics; LC/ESI-MS; Life Sciences; Lymphoma; Medical sciences; Medication; Melanoma; Molecular Structure; Pharmaceutical sciences; Pharmacology. Drug treatments; Quinoxalines - blood; Quinoxalines - chemistry; Quinoxalines - pharmacokinetics; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.200800668

Since several years, our group developed quinoxalinic compounds. Among the synthesized compounds, in the imidazo[1,2-a]quinoxaline series, EAPB0203 has shown interesting activities both on melanoma and lymphoma. The structure of EAPB0203 has been modulated and a new compound, EAPB0503, exhibits an in vitro cytotoxic activity on melanoma cancer cell line 7-9 times higher than EAPB0203. We validated an LC/ESI-MS method to simultaneously quantify EAPB0503 and its metabolite EAPB0603 in human and rat plasma. Chromatography was performed on a C8 Zorbax eclipse XDB column with a mobile phase consisting of acetronitrile and formate buffer gradient elution. LC-MS data were acquired in SIM mode at m/z 305, 291, and 303 for EAPB0503, EAPB0603, and the internal standard, respectively. The drug/internal standard peak area ratios were linked via quadratic relationships to concentrations (low range: 5-300 μg/L, high range: 100-1000 μg/L). The method is precise (precision, [less, not equals]14%) and accurate (recovery, 92-113%). Mean extraction efficiencies, >72% for each analyte, were obtained. The lower LOQs were 5 μg/L. This highly specific and sensitive method was successfully used to investigate plasma concentrations of EAPB0503 and EAPB0603 in a pharmacokinetic study carried out in rat and would also be useful in clinical trials at a later stage.