title: Development of a high-performance liquid chromatographic-mass spectrometric assay for the specific and sensitive quantification of Ro 64-0802, an anti-influenza drug, and its pro-drug, oseltamivir, in human and animal plasma and urine.
abstract: Oseltamivir phosphate (Ro 64-0796/002) is a pro-drug of the anti-influenza neuraminidase inhibitor, Ro 64-0802, and as Tamiflu, has been developed for the treatment of both A and B strains of the disease. This paper describes an HPLC-MS-MS assay for both compounds in plasma and urine which fulfils all of the criteria for a good analytical method. It is sensitive with limits of quantification of 1 and 10 ng/ml for the pro-drug and active neuraminidase inhibitor, respectively. It is both accurate and precise with typical coefficients of variation from some 5,000 quality control samples of approximately +/-3 and +/-6%, respectively. Extensive stability studies have demonstrated the absence of significant problems associated with the decomposition of either compound, although ex vivo hydrolysis of Ro 64-0796 to Ro 64-0802 in rodent plasma has to be prevented by the use of the esterase inhibitor, dichlorvos.
authors.affiliation: Roche Discovery Welwyn, Welwyn Garden City, Herts, UK.
authors.forename: H
authors.initials: H
authors.lastname: Wiltshire
authors.forename: B
authors.initials: B
authors.lastname: Wiltshire
authors.forename: A
authors.initials: A
authors.lastname: Citron
authors.forename: T
authors.initials: T
authors.lastname: Clarke
authors.forename: C
authors.initials: C
authors.lastname: Serpe
authors.forename: D
authors.initials: D
authors.lastname: Gray
authors.forename: W
authors.initials: W
authors.lastname: Herron
mesh_terms: D000081:Acetamides / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D000818:Animals; D000998:Antiviral Agents / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007251:Influenza, Human / Q000188:drug therapy; D013058:Mass Spectrometry; D053139:Oseltamivir; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000081:Acetamides; D000998:Antiviral Agents; C403132:Ro 64-0802; D053139:Oseltamivir