Improvement in HPLC separation of porphyrin isomers and application to biochemical diagnosis of porphyrias
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Pascale Macours
Abstract
Background: Identification of porphyrias relies on the measurement of different porphyrins in urine, feces and plasma. Separation of porphyrin isomers is essential for the differential diagnosis of some porphyrias.
Method: Separation of naturally occurring porphyrins was achieved on a Chromolith RP-18 column with fluorimetric detection using a methanol/ammonium acetate gradient mobile phase. Fecal and plasma porphyrins were extracted with acetonitrile and water at different pH values.
Results: Eight porphyrins including protoporphyrin eluted within 20min with good resolution of each of the I and III positional isomer pairs for standards, urine and plasma, and within 50min for feces. Improvement of the extraction method for fecal and plasmatic porphyrins resulted in high recovery (up to 89%) and reliable quantification of protoporphyrin.
Conclusions: The present RP-HPLC method is specific and efficient for routine analysis of porphyrins in human urine, feces and plasma.
Clin Chem Lab Med 2006;44:1433–40.
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©2006 by Walter de Gruyter Berlin New York
Articles in the same Issue
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Articles in the same Issue
- Initiation and progression of atherosclerosis – enzymatic or oxidative modification of low-density lipoprotein?
- Blood transfusions in athletes. Old dogmas, new tricks
- Molecular detection of tyrosinase transcripts in peripheral blood from patients with malignant melanoma: correlation of PCR sensitivity threshold with clinical and pathologic disease characteristics
- Increase in and clearance of cell-free plasma DNA in hemodialysis quantified by real-time PCR
- Lipoprotein lipase gene polymorphism at the PvuII locus and serum lipid levels in Guangxi Hei Yi Zhuang and Han populations
- Interpretation of cardiac troponin T behaviour in size-exclusion chromatography
- Point-of-care C-reactive protein testing in febrile children in general practice
- Improvement in HPLC separation of porphyrin isomers and application to biochemical diagnosis of porphyrias
- Measurement of late-night salivary cortisol with an automated immunoassay system
- Combining markers of nephrotoxicity and hepatotoxicity for improved monitoring and detection of chronic alcohol abuse
- Stone or stricture as a cause of extrahepatic cholestasis – do liver function tests predict the diagnosis?
- Insulin resistance and enhanced protein glycation in men with prehypertension
- Prevalence-dependent decision limits for the early detection of type 2 diabetes mellitus in venous blood, venous plasma and capillary blood during glucose challenge
- Analytical performance and clinical utility of the INNOTEST® PHOSPHO-TAU(181P) assay for discrimination between Alzheimer's disease and dementia with Lewy bodies
- Variations in assay protocol for the Dako cystatin C method may change patient results by 50% without changing the results for controls
- Approved IFCC recommendation on reporting results for blood glucose: International Federation of Clinical Chemistry and Laboratory Medicine Scientific Division, Working Group on Selective Electrodes and Point-of-Care Testing (IFCC-SD-WG-SEPOCT)
- National survey on the pre-analytical variability in a representative cohort of Italian laboratories
- 10% CV concentration for the fourth generation Roche cardiac troponin T assay derived from Internal Quality Control data
- Biological variation of non-SI traceable biological quantities: example of proteins
- Effect of tibolone therapy on lipids and coagulation indices
- Acknowledgement
- Contents Volume 44, 2006
- Author Index
- Subject Index
