Navegar

Colección

Datos Estadísticas

Artículo

La versión final de este artículo es de uso interno. El editor solo permite incluir en el repositorio el artículo en su versión post-print. Por favor, si usted la posee enviela a
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

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.

Registro:

Documento: Artículo
Título:A spectrophotometric method for estimating hemin in biological systems
Autor:Lombardo, M.E.; Araujo, L.S.; Ciccarelli, A.B.; Batlle, A.
Filiación:Centro de Investigaciones Sobre Porfirinas Y Porfirias-CIPYP (CONICET-FCEN, UBA), Ciudad Universitaria, Pabellón II, 1428 Buenos Aires, Argentina
Viamonte 1881, 10 A, CP-1056, Buenos Aires, Argentina
Palabras clave: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
Año:2005
Volumen:341
Número:2
Página de inicio:199
Página de fin:203
DOI: http://dx.doi.org/10.1016/j.ab.2004.11.002
Título revista:Analytical Biochemistry
Título revista abreviado:Anal. Biochem.
ISSN:00032697
CODEN:ANBCA
CAS:chloroform, 67-66-3; hemin, 16009-13-5; porphyrin, 24869-67-8; pyridine, 110-86-1; Chloroform, 67-66-3; Hemin, 16009-13-5; Serum Albumin, Bovine
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v341_n2_p199_Lombardo

Referencias:

  • Del, A.M., Batlle, C., Biosíntesis del Hemo (1997) Acta Bioquím. Clín. Latinoamericana Sup., 3. , Porfirias y Porfirinas. Aspectos Clínicos, bioquímicos y biologí a molecular Argentina
  • Pal, J.K., Joshi-Purandare, M., Dose-dependent differential effect of hemin on protein synthesis and cell proliferation in Leishmania donovani promastigotes cultured in vitro (2001) J. Biosci., 26, pp. 225-231
  • Chen, J.-J., London, I.M., Regulation of protein synthesis by heme-regulated eIF-2α kinase (1995) Trends Biochem. Sci., 20, pp. 105-108
  • Pal, J.K., Anand, S., Joseph, J., Association of Hsp90 with the heme-regulated eukaryotic initiation factor 2α kinase-a collaboration for regulating protein synthesis (1996) J. Biosci., 21, pp. 191-205
  • Chang, C.S., Chang, K.P., Heme requirement and acquisition by extracellular and intracellular stages of Leishmania mexicana amazoneusis (1985) Mol. Biochem. Parasitol., 16, pp. 267-276
  • Bauer, J., Fornnarino, J., High-performance liquid chromatographic analysis of "available" hemin in hematin solutions (1984) J. Chromatogr., 283, pp. 378-382
  • Genco, C.A., Odusanya, B.M., Brown, G., Binding and accumulation of hemin in Porphyromonas gingivalis are induced by hemin (1994) Infect. Inmmun., 62, pp. 2885-2892
  • Porra, R.J., Jones, O.T.G., Studies on ferrochelatase. 1. Assay and properties of ferrochelatase from a pig-liver mitochondrial extract (1963) Biochem. J., 87, pp. 181-185
  • Clezy, P.S., Morell, D.B., A spectroscopic study of haematin compounds in the Soret region (1963) Biochim. Biophys. Acta, 71, pp. 150-156
  • Lombardo, M.E., Araujo, L.S., Batlle, A., 5-Aminolevulinic acid synthesis in epimastigotes of Trypanosoma cruzi (2003) Int. J. Biochem. Cell Biol., 35, pp. 1263-1271
  • Lombardo, M.E., Araujo, L.S., Branca, A., Del, A.M., Batlle, C., A method for estimating glycine in the presence of excess glutamate with o-phthaldialdehyde (1990) Z. Naturforsch. C, 45, pp. 911-914

Citas:

---------- APA ----------
Lombardo, M.E., Araujo, L.S., Ciccarelli, A.B. & Batlle, A. (2005) . A spectrophotometric method for estimating hemin in biological systems. Analytical Biochemistry, 341(2), 199-203.
http://dx.doi.org/10.1016/j.ab.2004.11.002
---------- CHICAGO ----------
Lombardo, M.E., Araujo, L.S., Ciccarelli, A.B., Batlle, A. "A spectrophotometric method for estimating hemin in biological systems" . Analytical Biochemistry 341, no. 2 (2005) : 199-203.
http://dx.doi.org/10.1016/j.ab.2004.11.002
---------- MLA ----------
Lombardo, M.E., Araujo, L.S., Ciccarelli, A.B., Batlle, A. "A spectrophotometric method for estimating hemin in biological systems" . Analytical Biochemistry, vol. 341, no. 2, 2005, pp. 199-203.
http://dx.doi.org/10.1016/j.ab.2004.11.002
---------- VANCOUVER ----------
Lombardo, M.E., Araujo, L.S., Ciccarelli, A.B., Batlle, A. A spectrophotometric method for estimating hemin in biological systems. Anal. Biochem. 2005;341(2):199-203.
http://dx.doi.org/10.1016/j.ab.2004.11.002