title: Selective high-performance liquid chromatographic assay for itraconazole and hydroxyitraconazole in plasma from human immunodeficiency virus-infected patients.
abstract: A sensitive and selective reversed-phase liquid chromatographic assay for itraconazole and hydroxyitraconazole in human plasma has been developed and validated. Itraconazole and hydroxyitraconazole were extracted from the matrix using solid-phase extraction on a strong cation-exchange sorbent. All compounds were detected using fluorescence at 265 and 363 nm for excitation and emission, respectively. The assay has been validated over the range 10-1,000 ng/ml for both compounds, 10 ng/ml being the lower limit of quantification. Accuracies ranged from 104 to 113% for itraconazole and from 91 to 103% for hydroxyitraconazole. The intra-assay precisions were all below 9% for itraconazole and below 8% for hydroxyitraconazole. The selectivity has been evaluated with respect to all registered anti-human immunodeficiency virus (HIV) drugs and other potential co-medications and a few of their metabolites, commonly used by HIV-infected individuals. Both itraconazole and hydroxyitraconazole were stable under relevant conditions for HIV-inactivation and storage of samples. The applicability of the assay was demonstrated for samples collected from a treated HIV-infected patient.
authors.affiliation: Department of Pharmacy and Pharmacology, Slotervaart Hospital, Amsterdam, The Netherlands.
authors.forename: Cees H W
authors.initials: CH
authors.lastname: Koks
authors.forename: Rolf W
authors.initials: RW
authors.lastname: Sparidans
authors.forename: Georgette
authors.initials: G
authors.lastname: Lucassen
authors.forename: Kristel M L
authors.initials: KM
authors.lastname: Crommentuyn
authors.forename: Jos H
authors.initials: JH
authors.lastname: Beijnen
mesh_terms: D000935:Antifungal Agents / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015658:HIV Infections / Q000097:blood*; D006801:Humans; D017964:Itraconazole / Q000031:analogs & derivatives* / Q000097:blood* / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
chemical_list: D000935:Antifungal Agents; C084794:hydroxyitraconazole; D017964:Itraconazole