Selective high-performance liquid chromatographic assay for itraconazole and hydroxyitraconazole in plasma from human immunodeficiency virus-infected patients

J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Feb 5;767(1):103-10. doi: 10.1016/s0378-4347(01)00550-3.


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.

MeSH terms

  • Antifungal Agents / blood*
  • Antifungal Agents / pharmacokinetics
  • Chromatography, High Pressure Liquid / methods*
  • HIV Infections / blood*
  • Humans
  • Itraconazole / analogs & derivatives*
  • Itraconazole / blood*
  • Itraconazole / pharmacokinetics
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence


  • Antifungal Agents
  • hydroxyitraconazole
  • Itraconazole