Sensitive isocratic liquid chromatographic assay for the determination of 5,10,15,20-tetra(m-hydroxyphenyl)chlorin in plasma and tissue with electrochemical detection.
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.
Barberi-Heyob
M
M
Laboratoire de Recherche en Oncologie, Centre Alexis Vautrin, Vandoeuvre-les-Nancy, France.
Rezzoug
H
H
Merlin
J L
JL
Guillemin
F
F
D000230:Adenocarcinoma / Q000737:chemistry; D000818:Animals; D000970:Antineoplastic Agents / Q000032:analysis* / Q000097:blood; D002294:Carcinoma, Squamous Cell / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003110:Colonic Neoplasms / Q000737:chemistry; D004563:Electrochemistry; D005260:Female; D006801:Humans; D008652:Mesoporphyrins / Q000032:analysis* / Q000097:blood; D051379:Mice; D008819:Mice, Nude; D017319:Photosensitizing Agents / Q000032:analysis* / Q000097:blood; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000970:Antineoplastic Agents; D008652:Mesoporphyrins; D017319:Photosensitizing Agents; C072269:temoporfin