A spectrophotometric method for estimating hemin in biological systems
Hemin chlorides exhibit two absorption maxima in the Soret region, one at about 360-380 nm (S' band) and the other between 400 and 430 nm (S band). We present here a simple and fast spectrophotometric assay to determine concentration of hemin between 1.15 and 9.20 μM employing the Soret region...
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2005
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v341_n2_p199_Lombardo http://hdl.handle.net/20.500.12110/paper_00032697_v341_n2_p199_Lombardo |
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paper:paper_00032697_v341_n2_p199_Lombardo2023-06-08T14:24:04Z A spectrophotometric method for estimating hemin in biological systems Lombardo, María Elisa Araujo, Lidia Susana Ciccarelli, Alejandra Beatriz Batlle, Alcira María del Carmen Hemin extraction Hemin quantification Pyridine hemochromogen method S' Soret band chloroform chromogenic substrate hemin porphyrin pyridine absorption acidity amino acid analysis article correlation analysis extract extraction linear system priority journal quantitative assay recording reproducibility sensitivity analysis spectrophotometry Animals Chloroform Hemin Saccharomyces cerevisiae Serum Albumin, Bovine Spectrophotometry Spectrophotometry, Ultraviolet Trypanosoma cruzi Hemin chlorides exhibit two absorption maxima in the Soret region, one at about 360-380 nm (S' band) and the other between 400 and 430 nm (S band). We present here a simple and fast spectrophotometric assay to determine concentration of hemin between 1.15 and 9.20 μM employing the Soret region (S' band) as a reference. In this method the hemin is quantitatively extracted from biological materials by acidified chloroform. By recording the absorbance of the chloroform extract at its maximum peak at 388, 450, and 330 nm and applying the correction formula Ac = 2A388 - (A 450 + A330), a very good linear correlation between the Ac and the concentration of hemin is attained. The method can be used to estimate hemin in the presence of protein (0.06-5.00 mg/ml) and porphyrin (0.19-2.97 μM). Compared with the pyridine hemochromogen method, the assay reported here is highly reproducible, with 15- to 30-fold more sensitivity, and it allows the quantification of four times lower hemin concentrations. © 2004 Elsevier Inc. All rights reserved. Fil:Lombardo, M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Araujo, L.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ciccarelli, A.B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Batlle, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2005 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v341_n2_p199_Lombardo http://hdl.handle.net/20.500.12110/paper_00032697_v341_n2_p199_Lombardo |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Hemin extraction Hemin quantification Pyridine hemochromogen method S' Soret band chloroform chromogenic substrate hemin porphyrin pyridine absorption acidity amino acid analysis article correlation analysis extract extraction linear system priority journal quantitative assay recording reproducibility sensitivity analysis spectrophotometry Animals Chloroform Hemin Saccharomyces cerevisiae Serum Albumin, Bovine Spectrophotometry Spectrophotometry, Ultraviolet Trypanosoma cruzi |
| spellingShingle |
Hemin extraction Hemin quantification Pyridine hemochromogen method S' Soret band chloroform chromogenic substrate hemin porphyrin pyridine absorption acidity amino acid analysis article correlation analysis extract extraction linear system priority journal quantitative assay recording reproducibility sensitivity analysis spectrophotometry Animals Chloroform Hemin Saccharomyces cerevisiae Serum Albumin, Bovine Spectrophotometry Spectrophotometry, Ultraviolet Trypanosoma cruzi Lombardo, María Elisa Araujo, Lidia Susana Ciccarelli, Alejandra Beatriz Batlle, Alcira María del Carmen A spectrophotometric method for estimating hemin in biological systems |
| topic_facet |
Hemin extraction Hemin quantification Pyridine hemochromogen method S' Soret band chloroform chromogenic substrate hemin porphyrin pyridine absorption acidity amino acid analysis article correlation analysis extract extraction linear system priority journal quantitative assay recording reproducibility sensitivity analysis spectrophotometry Animals Chloroform Hemin Saccharomyces cerevisiae Serum Albumin, Bovine Spectrophotometry Spectrophotometry, Ultraviolet Trypanosoma cruzi |
| description |
Hemin chlorides exhibit two absorption maxima in the Soret region, one at about 360-380 nm (S' band) and the other between 400 and 430 nm (S band). We present here a simple and fast spectrophotometric assay to determine concentration of hemin between 1.15 and 9.20 μM employing the Soret region (S' band) as a reference. In this method the hemin is quantitatively extracted from biological materials by acidified chloroform. By recording the absorbance of the chloroform extract at its maximum peak at 388, 450, and 330 nm and applying the correction formula Ac = 2A388 - (A 450 + A330), a very good linear correlation between the Ac and the concentration of hemin is attained. The method can be used to estimate hemin in the presence of protein (0.06-5.00 mg/ml) and porphyrin (0.19-2.97 μM). Compared with the pyridine hemochromogen method, the assay reported here is highly reproducible, with 15- to 30-fold more sensitivity, and it allows the quantification of four times lower hemin concentrations. © 2004 Elsevier Inc. All rights reserved. |
| author |
Lombardo, María Elisa Araujo, Lidia Susana Ciccarelli, Alejandra Beatriz Batlle, Alcira María del Carmen |
| author_facet |
Lombardo, María Elisa Araujo, Lidia Susana Ciccarelli, Alejandra Beatriz Batlle, Alcira María del Carmen |
| author_sort |
Lombardo, María Elisa |
| title |
A spectrophotometric method for estimating hemin in biological systems |
| title_short |
A spectrophotometric method for estimating hemin in biological systems |
| title_full |
A spectrophotometric method for estimating hemin in biological systems |
| title_fullStr |
A spectrophotometric method for estimating hemin in biological systems |
| title_full_unstemmed |
A spectrophotometric method for estimating hemin in biological systems |
| title_sort |
spectrophotometric method for estimating hemin in biological systems |
| publishDate |
2005 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v341_n2_p199_Lombardo http://hdl.handle.net/20.500.12110/paper_00032697_v341_n2_p199_Lombardo |
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