Measurement of the novel antitumor agent 17-(allylamino)-17-demethoxygeldanamycin in human plasma by high-performance liquid chromatography

Elizabeth B. Agnew, Richard H. Wilson, Jean L. Grem, Leonard Neckers, Daoqin Bi, Chris H. Takimoto

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

A sensitive HPLC assay has been developed to determine the concentration of 17-(allylamino)-17-demethoxygeldanamycin (AAG) in human plasma over the concentration range of 12.5 to 2500 nM (7.33 to 1465 ng/mL). After the addition of 1000 nM geldanamycin as the internal standard, 1 mL samples of human plasma were subjected to solid-phase extraction, via Bond-Elut C18 cartridges, followed by analysis using an isocratic reversed-phase HPLC assay with UV detection. A Phenomenex Kingsorb, 3 micron, C18, 150×4.60 mm column and a Phenomenex Security Guard pre-column, C18 (ODS, Octadecyl), were used to achieve separation. AAG and GM were monitored at 334 and 308 nm, respectively, on a Hewlett-Packard 1050 Diode-Array Detector. The mobile phase, run at a flow-rate of 1 mL/min, was composed of 50% (v/v) 25 mM sodium phosphate (pH 3.00) with 10 mM triethylamine and 50% acetonitrile. HPLC effectively resolved AAG with retention times of 14.60±0.54 min and the internal standard geldanamycin at 10.72±0.38 min (n=15). This assay was able to measure plasma concentrations of AAG, the lower limit of quantitation being 12.5 nM, at a starting dose of 10 mg/m2 infused intravenously over 1 h in a Phase I clinical trial in adult patients with solid tumors.

Original languageEnglish (US)
Pages (from-to)237-243
Number of pages7
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume755
Issue number1-2
DOIs
StatePublished - May 5 2001

Keywords

  • 17-(Allylamino)-17-demethoxygeldanamycin

ASJC Scopus subject areas

  • Chemistry(all)

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