Development and standardisation of a fluorimetric method for the quantitation of total faecal porphyrins
Cardiff Metropolitan University
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The porphyrias are a group of rare disorders that are caused by a genetic defect of the haem biosynthetic pathway which lead to the overproduction of the haem precursors. The clinical features of the porphyrias are associated with specific porphyrin patterns and the overproduction, accumulation and excretion of the porphyrin precursors. The Cardiff Porphyria Service currently performs quantitation of total porphyrins in faeces by a method using spectrophotometric detection. This method is subject to interference, lacks standardisation and is subject to background absorbance due to impurities. Fluorimetry offers an alternative method of detection which may be more sensitive and specific. A fluorimetric method for quantitation of faecal porphyrins was devised based on that of Pudek et al and compared with the existing method (Lockwood et al, 1985 & Pudek et al, 1991). The fluorimetric method demonstrates no advantage over the existing method both practically and in terms of assay performance. Both, extremely manual, methods exhibit broad variation over the assay range. For these reasons the original spectrophotometric method remains the assay of choice.
MSc Biomedical Science
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