Sensitive determination of itraconazole and its active metabolite in human plasma by column-switching high-performance liquid chromatography with ultraviolet detection

Tsukasa Uno, Mikiko Shimizu, Kazunobu Sugawara, Tomonori Tateishi

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

A simple and sensitive column-switching high-performance liquid chromatographic method for the simultaneous determination of itraconazole (ITZ) and its active metabolite, hydroxyitraconazole (HIT) in human plasma is described. ITZ, HIT, and an internal standard, R051012, were extracted from 1 mL of alkalinized plasma sample using n-heptane-chloroform (60:40, vol/vol). The extract was injected onto column I (TSK precolumn BSA-ODS/S, 5 μm, 10×4.6 mm ID) for clean-up and column II (Develosil C8-5 column, 5 μm, 150×4.6 mm ID) for separation. The mobile phase consisted of phosphate buffer-acetonitrile (68:32 vol/vol, pH 6.0) for clean-up and phosphate buffer-acetonitrile (35:65 vol/vol, pH 6.0) for separation. The peaks were monitored with an ultraviolet detector set at a wavelength of 263 nm, and total time for chromatographic separation was about 24 minutes. The validated concentration ranges of this method were 3 to 500 ng/mL for ITZ and 3 to 1000 ng/mL for HIT. Mean recoveries were 59.7% for ITZ and 72.8% for HIT. Intraday and interday coefficients of variation were less than 4.6% and 5.0% for ITZ, and 4.6% and 4.9% for HIT at the different concentrations. The limit of quantification was 3 ng/mL for both ITZ and HIT. This method was suitable for therapeutic drug monitoring of ITZ and HIT, and was applied to pharmacokinetic studies in human volunteers.

Original languageEnglish
Pages (from-to)526-531
Number of pages6
JournalTherapeutic Drug Monitoring
Volume28
Issue number4
DOIs
Publication statusPublished - 2006 Aug
Externally publishedYes

Fingerprint

Plasma (human)
Itraconazole
High performance liquid chromatography
Metabolites
High Pressure Liquid Chromatography
Buffers
Phosphates
Ultraviolet detectors
Pharmacokinetics
Drug Monitoring
Chloroform
hydroxyitraconazole
Volunteers
Plasmas
Recovery
Wavelength
Monitoring
Liquids
Pharmaceutical Preparations

Keywords

  • Columnswitching
  • HPLC
  • Hydroxyitraconazole
  • Itraconazole
  • Therapeutic drug monitoring

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis
  • Pharmacology
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology (medical)
  • Public Health, Environmental and Occupational Health

Cite this

Sensitive determination of itraconazole and its active metabolite in human plasma by column-switching high-performance liquid chromatography with ultraviolet detection. / Uno, Tsukasa; Shimizu, Mikiko; Sugawara, Kazunobu; Tateishi, Tomonori.

In: Therapeutic Drug Monitoring, Vol. 28, No. 4, 08.2006, p. 526-531.

Research output: Contribution to journalArticle

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abstract = "A simple and sensitive column-switching high-performance liquid chromatographic method for the simultaneous determination of itraconazole (ITZ) and its active metabolite, hydroxyitraconazole (HIT) in human plasma is described. ITZ, HIT, and an internal standard, R051012, were extracted from 1 mL of alkalinized plasma sample using n-heptane-chloroform (60:40, vol/vol). The extract was injected onto column I (TSK precolumn BSA-ODS/S, 5 μm, 10×4.6 mm ID) for clean-up and column II (Develosil C8-5 column, 5 μm, 150×4.6 mm ID) for separation. The mobile phase consisted of phosphate buffer-acetonitrile (68:32 vol/vol, pH 6.0) for clean-up and phosphate buffer-acetonitrile (35:65 vol/vol, pH 6.0) for separation. The peaks were monitored with an ultraviolet detector set at a wavelength of 263 nm, and total time for chromatographic separation was about 24 minutes. The validated concentration ranges of this method were 3 to 500 ng/mL for ITZ and 3 to 1000 ng/mL for HIT. Mean recoveries were 59.7{\%} for ITZ and 72.8{\%} for HIT. Intraday and interday coefficients of variation were less than 4.6{\%} and 5.0{\%} for ITZ, and 4.6{\%} and 4.9{\%} for HIT at the different concentrations. The limit of quantification was 3 ng/mL for both ITZ and HIT. This method was suitable for therapeutic drug monitoring of ITZ and HIT, and was applied to pharmacokinetic studies in human volunteers.",
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