| Microanalysis of non-heme iron in animal tissues. | |
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MedLine Citation:
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PMID: 15026210 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The method recommended by the Iron Panel of the International Committee for the Standardization in Haematology for measurement of serum iron was adapted for measurement of non-heme iron in animal tissues. The method developed was designed specifically to facilitate measurement of non-heme iron using as little as 10 mg of tissue, in a final reaction volume of 60 microl. In this assay, tissue homogenates are treated with hydrochloric acid and trichloroacetic acid and heated at 95 degrees C. Non-heme iron is released and protein is precipitated. Following centrifugation, iron in the supernatant is reacted with ferrozine in the presence of the reducing agent thioglycolic acid, and the complex is quantified by spectrophotometry. The method was validated by analysis of two Standard Reference Materials (bovine liver), comparing results of this assay against certified values and concentrations determined by flame atomic absorption spectrometry following acid digestion. Results using this method for analysis of non-heme iron in guinea pig tissues (liver, kidney and heart) compared favorably with those obtained using micro-scale adaptations of three published reference methods. The new method was more sensitive, required less time, and was less cumbersome than the three published methods to which it was compared. |
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Authors:
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Charles J Rebouche; Cari L Wilcox; John A Widness |
Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Journal of biochemical and biophysical methods Volume: 58 ISSN: 0165-022X ISO Abbreviation: J. Biochem. Biophys. Methods Publication Date: 2004 Mar |
Date Detail:
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Created Date: 2004-03-17 Completed Date: 2004-11-30 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 7907378 Medline TA: J Biochem Biophys Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 239-51 Citation Subset: IM |
Affiliation:
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Department of Pediatrics, University of Iowa, Iowa City, IA 52242, USA. charles-rebouche@uiowa.edu |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Ferrozine / chemistry Guinea Pigs Heme / chemistry Hydrochloric Acid / chemistry Iron / analysis*, chemistry Iron Chelating Agents / chemistry Kidney / chemistry Liver / chemistry Myocardium / chemistry Reference Standards Sensitivity and Specificity Spectrophotometry, Atomic / methods* Thioglycolates / chemistry Trichloroacetic Acid / chemistry |
| Grant Support | |
ID/Acronym/Agency:
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P01 HL46925/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Iron Chelating Agents; 0/Thioglycolates; 14875-96-8/Heme; 28048-33-1/Ferrozine; 68-11-1/2-mercaptoacetate; 7439-89-6/Iron; 76-03-9/Trichloroacetic Acid; 7647-01-0/Hydrochloric Acid |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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