Dual-wavelength excitation for fluorescence-based quantification of zinc protoporphyrin IX and protoporphyrin IX in whole blood
| Autor: | Herbert Stepp, Christian Gruber, Georg Hennig, Stephan Dittmar, Gary M. Brittenham, Michael Vogeser, Ronald Sroka |
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| Rok vydání: | 2013 |
| Předmět: |
Chromatography
Protoporphyrin IX Zinc protoporphyrin General Engineering General Physics and Astronomy General Chemistry High-performance liquid chromatography Fluorescence General Biochemistry Genetics and Molecular Biology Fluorescence spectroscopy chemistry.chemical_compound chemistry Biochemistry Erythropoiesis General Materials Science Heme Whole blood |
| Zdroj: | Journal of Biophotonics. 7:514-524 |
| ISSN: | 1864-063X |
| Popis: | Quantification of erythrocyte zinc protoporphyrin IX (ZnPP) and protoporphyrin IX (PPIX), individually or jointly, is useful for the diagnostic evaluation of iron deficiency, iron-restricted erythropoiesis, lead exposure, and porphyrias. A method for simultaneous quantification of ZnPP and PPIX in unwashed blood samples is described, using dual-wavelength excitation to effectively eliminate background fluorescence from other blood constituents. In blood samples from 35 subjects, the results of the dual-wavelength excitation method and a reference high performance liquid chromatography (HPLC) assay were closely correlated both for ZnPP (rs = 0.943, p < 0.0001; range 37-689 μmol ZnPP/mol heme, 84-1238 nmol/L) and for PPIX (rs = 0.959, p < 0.0001; range 42-4212 μmol PPIX/mol heme, 93-5394 nmol/L). In addition, for ZnPP, the proposed method is compared with conventional single-wavelength excitation and with commercial front-face fluorimetry of washed erythrocytes and whole blood. We hypothesize that dual-wavelength excitation fluorimetry will provide a new approach to the suppression of background fluorescence in blood and tissue measurements of ZnPP and PPIX. |
| Databáze: | OpenAIRE |
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