Journal
BIOMEDICAL CHROMATOGRAPHY
Volume 20, Issue 12, Pages 1277-1282Publisher
WILEY
DOI: 10.1002/bmc.624
Keywords
hematoporphyrin monomethyl ether; photodynamic therapy; pharmacokinetics; high-performance liquid chromatography; fluorescence detection
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A rapid, sensitive, precise and specific method for determination of hematoporphyrin monomethyl ether (HMME), a novel photodynamic therapy (PDT) drug, was developed and validated using high-performance liquid chromatography (HPLC) with fluorescence detection. HMME was isolated from the plasma by a single-step liquid-liquid extraction with ethyl acetate. The analyte and internal standard fluorescein were baseline separated on a Diamonsil C-18 analytical column (4.6 x 150 mm, 5 mu m) and analyzed using a fluorescence detector with the excitation and emission wavelengths set at 395 and 613 run, respectively. The method was linear in the concentration range 0.025-5 mu g/mL with a lower limit of quantitation (LLOQ) of 10 ng/mL. The inter and intra-day accuracies and precisions were all within 10% and the mean recoveries of HMME and fluorescein were 95 +/- 3.7 and 90 +/- 2.3%, respectively. The analyte was stable during all sample storage, preparation and analysis periods. This method was successfully applied to a pharmacokinetic study after a single-dose intravenous administration of HMME (5 mg/kg) to beagle dogs. This method was reproducible and sensitive enough for the pharmacokinetic study of HMME. Based on the results of the pharmacokinetic study, we suggest that a rather long light-avoiding time is essential for patients under HMME therapy. Copyright (c) 2006 John Wiley & Sons, Ltd.
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