Sensitive isocratic liquid chromatographic assay for the determination of 5,10,15,20-tetra(m-hydroxyphenyl)chlorin in plasma and tissue with electrochemical detection.
Fiche publication
Date publication
janvier 1997
Journal
Journal of chromatography. B, Biomedical sciences and applications
Auteurs
Membres identifiés du Cancéropôle Est :
Pr BARBERI-HEYOB Muriel, Pr MERLIN Jean-Louis
Tous les auteurs :
Barberi-Heyob M, Rezzoug H, Merlin JL, Guillemin F
Lien Pubmed
Résumé
A simple extraction procedure and a sensitive high-performance liquid chromatographic (HPLC) method are described for the determination of the photodynamic therapeutic agent 5,10,15,20-tetra(m-hydroxyphenyl)chlorin (mTHPC) in plasma and tumour tissue. Reversed-phase high-performance liquid chromatography was performed on a C18 column (70 x 4.6 mm I.D.) with a mobile phase of 0.01 M potassium dihydrogenphosphate buffer, pH 2.5-acetonitrile (55:45, v/v) and a coulometric detection (+0.80 V). The mean recoveries of mTHPC in the concentration ranges (5-2000 and 10-1000 ng/ml) were 90 and 89% for plasma and tumour samples, respectively. The procedure for plasma and tissue preparation involved solvent precipitation using methanol combined with ammonia solution and dimethyl sulphoxide (4, 0.2, 0.1, v/v/v) and (2, 0.1, 0.1, v/v/v) for plasma and tissue, respectively. For mTHPC at concentrations ranging from 5 to 2000 ng/ml, the within-day relative standard deviations, based on triplicate determinations were less than 8% and the between-day relative standard deviations calculated by performing extraction procedure of plasma samples on three different days ranged from 3 to 18%. This highly sensitive method, 5 and 10 ng/ml for plasma and tissue respectively, was applied successfully to the determination of mTHPC in mouse tumours for pharmacokinetic studies.
Mots clés
Adenocarcinoma, chemistry, Animals, Antineoplastic Agents, analysis, Carcinoma, Squamous Cell, chemistry, Chromatography, High Pressure Liquid, methods, Colonic Neoplasms, chemistry, Electrochemistry, Female, Humans, Mesoporphyrins, analysis, Mice, Mice, Nude, Photosensitizing Agents, analysis, Reproducibility of Results, Sensitivity and Specificity
Référence
J. Chromatogr. B Biomed. Sci. Appl.. 1997 Jan;688(2):331-8