Red cell zinc protoporphyrin and protoporphyrin by HPLC with fluorescence detection

Authors

  • E. Rossi,

    Corresponding author
    1. University Department of Clinical Biochemistry, Queen Elizabeth II Medical Centre, Nedlands 6009, Australia
    • University Department of Clinical Biochemistry, Queen Elizabeth II Medical Centre, Nedlands 6009, Australia
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  • P. Garcia-Webb

    1. University Department of Clinical Biochemistry, Queen Elizabeth II Medical Centre, Nedlands 6009, Australia
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Abstract

A high performance liquid chromatographic (HPLC) method was developed for the determination of zinc protoporphyrin (ZnPP) and protoporphyrin (PP) in whole blood. After adding the blood to dilute acetic acid, ZnPP and PP were extracted with dimethyl sulfoxide–acetone containing mesoporphyrin as internal standard. Following evaporation of the acetone, the haemin-free extract was analysed by HPLC. ZnPP and PP were separated on a reversed-phase column and quantitated by measuring fluorescence peak areas. The extraction method is simple, and applicable to batch analysis, and the HPLC separation is rapid and repoducible. The coefficient of variation for ZnPP was 5.6% and 3.3% for total red cell porphyrin levels of 3.5 and 10.2 μmol per litre RBC respectively. Results are discussed in patients with erythrohepatic protoporphyria, lead exposure, iron-deficiency and non-specifically elevated total red cell porphyrins.

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