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Updated: May 31, 2026

Porphyrin-Modified Beads for Use as Compensation Controls in Flow Cytometry
Published on: March 24, 2023
Ultra high-performance liquid chromatography of porphyrins
Christopher M Benton1, Chang Kee Lim, Harald J Ritchie
1Clinical Biochemistry, King's College Hospital, Denmark Hill, London, SE5 9RS, UK.
Abstract:
An ultra high-performance liquid chromatographic (UHPLC) system was developed and optimized for the separation of porphyrins of clinical interest. Optimum conditions for the simultaneous separation of uroporphyrin, hepta-, hexa-, penta-carboxylic acid porphyrins and coproporphyrin and their type I and III isomers on a Thermo Hypersil BDS C18 column (2.4 µm particle size, 100 × 2.1 mm i.d.) using a gradient elution with 10% (v/v) acetonitrile in 1.0 m ammonium acetate buffer (pH 5.16) and 10% (v/v) acetonitrile in methanol at a flow-rate of 0.4 mL/min. The effect of mobile phase buffer molarity on the sensitivity of fluorescence detection and resolution of porphyrin isomers was investigated. The method was successfully applied to the analysis of porphyrins extracted from the urine and faeces of patients with various human porphyrias.
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