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Development and validation of a sensitive quantification method for hematoporphyrin monomethyl ether in plasma using high‐performance liquid chromatography with fluorescence detection
Author(s) -
Li Peng,
Sun Jianguo,
Huang Chenrong,
Pan GuoYu,
Xu Meijuan,
Li Jing,
Wang Guangji,
Tao Jining
Publication year - 2006
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.624
Subject(s) - chemistry , chromatography , analyte , pharmacokinetics , high performance liquid chromatography , detection limit , fluorescence , beagle , fluorescence spectroscopy , pharmacology , medicine , physics , quantum mechanics
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 × 150 mm, 5 µm) and analyzed using a fluorescence detector with the excitation and emission wavelengths set at 395 and 613 nm, respectively. The method was linear in the concentration range 0.025–5 µg/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 © 2006 John Wiley & Sons, Ltd.