The quantitative determination of Photofrin and Polyhaematoporphyrin in plasma: pitfalls and inaccuracies.

An accurate and relatively rapid fluorometric assay for Photofrin, or our own preparation Polyhaematoporphyrin (PHP), in plasma has been developed. This method takes into account the significant proportion of hydrolysis-resistant material now known to be present in these sensitizers, which has undoubtedly led to the inaccurate assessment of these drugs and other preparations of haematoporphyrin derivative (HPD) in numerous studies. The method was devised to allow for incomplete hydrolysis by calibration with plasma samples to which known amounts of photosensitizer were added in vitro. It was then compared with an "absolute" method in which plasma from animals that had received 14C-labelled drug was subjected to radioactivity assay. The two approaches gave almost identical results. The calibration method is applicable to the determination of any HPD-derived drug from patient or animal studies. As an example of its use in the present study, it was applied to the determination of the pharmacokinetics of PHP in the rat.