The rapid determination of erythrocyte porphyrins using reversed-phase high performance liquid chromatography

Clin Chim Acta. 1981 Jul 1;113(3):253-65. doi: 10.1016/0009-8981(81)90279-5.

Abstract

The extraction, separation and quantification of free acid erythrocyte porphyrins are described. The porphyrins are extracted from whole blood using 3:1 ethyl acetate/acetic acid (v/v) and separated using the reversed-phase mode of high performance liquid chromatography (RPLC). The compounds are detected on-line spectrofluorometrically using an excitation wavelength of 404 nm and an emission cut-off filter of 550 nm. Excellent resolution of the porphyrin free acids is achieved in less than 6 min. The analytical recoveries for protoporphyrin IX and zinc protoporphyrin IX were greater than 90% with relative standard deviations for day-to-day analysis under 6%.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetates
  • Acetic Acid
  • Chromatography, High Pressure Liquid
  • Coproporphyrins / blood
  • Erythrocytes / analysis*
  • Humans
  • Mesoporphyrins / blood
  • Porphyrins / blood*
  • Porphyrins / isolation & purification
  • Protoporphyrins / blood

Substances

  • Acetates
  • Coproporphyrins
  • Mesoporphyrins
  • Porphyrins
  • Protoporphyrins
  • mesoporphyrin IX
  • coproporphyrin I
  • ethyl acetate
  • protoporphyrin IX
  • Acetic Acid