Determination of 5,10,15,20‐ tetra‐ (m‐hydroxyphenyl)chlorin in human plasma by high performance liquid chromatography

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Abstract
A high performance liquid chromatographic method for the determination of the photodynamic chemotherapeutic agent 5,10,15,20‐tetra(m‐hydroxyphenyl)chlorin (m‐THPC) in human plasma following intravenous infusion is described. The procedure involves extraction of the drug in plasma with methanol/dimethyl sulphoxide (4:1 v/v) containing 5,10,15,20‐tetra(p‐hydroxyphenyl)clorin as the internal standard and separation on a C18 reversed phase column with acetonitrile:0.1% trifluoroacetic acid (77:23 v/v) as the mobile phase. The drug was detected specifically and sensitively at its absorption maximum of 423 nm with a detection limit of 15 ng/mL (signal‐to‐noise ratio of 5). The intra‐ and inter‐assay coefficients of variation (CV) on analysis of a plasma spiked with m‐THPC (1 μg/mL) were 2.3 and 3.4% (n=6), respectively.