High-pressure liquid chromatography of plasma free acid porphyrins

Longas, Marı́a O.; Poh–Fitzpatrick, Maureen B.

doi:10.1016/0003-2697(80)90074-3

Cited by 35 publications

(5 citation statements)

References 18 publications

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“…Porphyrin concentrations were determined by a method modified from that of Longas and Poh-Fitzpatrick (1980). The porphyrin extraction efficiency of this assay has previously been calculated as 90% (Poh-Fitzpatrick and DeLeo, 1977).…”

Section: Cells and Cell Culturementioning

confidence: 99%

Porphyrin accumulation induced by 5-aminolaevulinic acid esters in tumour cells growing in vitro and in vivo

et al. 2002

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The ability of 5-aminolaevulinic acid and some of its esterified derivatives to induce porphyrin accumulation has been examined in CaNT murine mammary carcinoma cells growing in culture and as tumours in vivo. Topical or intravenous administration of 5-aminolaevulinic acid-esters to mice bearing subcutaneous tumours produced lower porphyrin levels in the tumour than an equimolar dose of 5-aminolaevulinic acid. Reducing the dose of intravenous hexyl-or benzyl-ALA and topical hexyl-5-aminolaevulinic acid resulted in a dose-dependent reduction in porphyrin accumulation. A number of normal tissues accumulated higher concentrations of porphyrins than tumour tissue following intravenous administration of 5-aminolaevulinic acid-esters. Esterase activity in these normal tissues was greater than that in tumour tissue. In contrast to the situation in vivo, all of the 5-aminolaevulinic acid-esters examined were at least as effective as 5-aminolaevulinic acid when applied to cloned CaNT cells in vitro, with the drug concentration required for maximum porphyrin accumulation varying with ester chain-length. Tumour cells growing in culture released esterase activity into the medium. These findings suggest that the efficacy of 5-aminolaevulinic esters may vary depending on the esterase activity of the target tissue, and suggest caution when interpreting the findings of in vitro studies using these and similar prodrugs.

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Section: Cells and Cell Culturementioning

confidence: 99%

Porphyrin accumulation induced by 5-aminolaevulinic acid esters in tumour cells growing in vitro and in vivo

et al. 2002

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“…Measuring coproporphyrin concentrations with this method yields better results because the ratio of fluorescence intensities at 397 nm and 409 nm excitation is larger for coproporphyrin than for uroporphyrin. Taking only into account concentrations of 8 ng/ ml or higher, as indicated for acute porphyrias [8], the standard deviation improves to 3.7% and 10.0 % for determination of coproporphyrin-III and uroporphyrin-III, respectively. This potentially allows for a very exact analysis of uroporphyrin-III and coproporphyrin-III concentrations with a very fast and cheap method.…”

Section: Discussionmentioning

confidence: 97%

“…For AIP, uroporphyrin is elevated up to 25 ng per ml and coproporphyrin up to 43 ng per ml. [7,8] In this study, the chosen concentrations of 2 -16 ng per ml for the mixture are on the lower end of this scale and therefore the results are applicable to medical screening with real plasma samples. Comparing the measured concentrations with the initially adjusted ones, on average, 91 % of coproporphyrin-III and 118 % of uroporphyrin were recovered by the evaluation with a standard deviation of 10.3 % for coproporphyrin-III and of 43.8 % for uroporphyrin-III.…”

Section: Discussionmentioning

confidence: 99%

Rapid screening test for porphyria diagnosis using fluorescence spectroscopy

et al. 2015

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Porphyrias are rare genetic metabolic disorders, which result from deficiencies of enzymes in the heme biosynthesis pathway. Depending on the enzyme defect, different types of porphyrins and heme precursors accumulate for the different porphyria diseases in erythrocytes, liver, blood plasma, urine and stool. Patients with acute hepatic porphyrias can suffer from acute neuropathic attacks, which can lead to death when undiagnosed, but show only unspecific clinical symptoms such as abdominal pain. Therefore, in addition to chromatographic methods, a rapid screening test is required to allow for immediate identification and treatment of these patients. In this study, fluorescence spectroscopic measurements were conducted on blood plasma and phantom material, mimicking the composition of blood plasma of porphyria patients. Hydrochloric acid was used to differentiate the occurring porphyrins (uroporphyrin-III and coproporphyrin-III) spectroscopically despite their initially overlapping excitation spectra. Plasma phantom mixtures were measured using dual wavelength excitation and the corresponding concentrations of uroporphyrin-III and coproporphyrin-III were determined. Additionally, three plasma samples of porphyria patients were examined and traces of coproporphyrin-III and uroporphyrin-III were identified. This study may therefore help to establish a rapid screening test method with spectroscopic differentiation of the occurring porphyrins, which consequently allows for the distinction of different porphyrias. This may be a valuable tool for clinical porphyria diagnosis and rapid or immediate treatment.

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“…The excitation and emission wavelengths are usually set to around 395–420 and 580–620 nm, respectively. Early methods separated porphyrins using ion‐exchange (Evans et al ., ) and normal‐phase chromatography (Slavin et al ., ; Longas and Poh‐Fitzpatrick, ). This was soon replaced by reversed‐phase chromatography owing to its improved efficiency.…”

Section: Porphyrinsmentioning

confidence: 99%

Liquid chromatography and mass spectrometry of haem biosynthetic intermediates: a review

Benton

Lim

2012

Biomedical Chromatography

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This article discusses the separation, analysis and characterisation of intermediates and oxidative by-products of the haem biosynthetic pathway by liquid chromatography and mass spectrometry. Techniques reviewed include high-performance liquid chromatography, ultra-high-performance liquid chromatography, capillary electrophoresis, ion mobility spectrometry, mass spectrometry and tandem mass spectrometry. The emphasis was on the analysis of biological and clinical samples.

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High-pressure liquid chromatography of plasma free acid porphyrins

Cited by 35 publications

References 18 publications

Porphyrin accumulation induced by 5-aminolaevulinic acid esters in tumour cells growing in vitro and in vivo

Porphyrin accumulation induced by 5-aminolaevulinic acid esters in tumour cells growing in vitro and in vivo

Rapid screening test for porphyria diagnosis using fluorescence spectroscopy

Liquid chromatography and mass spectrometry of haem biosynthetic intermediates: a review

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