Document Detail

Adventitious redox-active metals in Krebs-Henseleit buffer can contribute to Langendorff heart experimental results.
MedLine Citation:
PMID:  8089856     Owner:  NLM     Status:  MEDLINE    
Adventitious redox-active metals in Krebs-Henseleit buffer exhibit a significant enhancement of damage to isolated rat hearts. Using atomic absorption spectroscopy, it was determined that Krebs-Henseleit buffer contains substantial amounts of contaminating iron and copper. Significant copper contamination was found in ACS Reagent grade sodium chloride and sodium bicarbonate; iron contamination in sodium chloride, potassium chloride, sodium bicarbonate, and calcium chloride. Chelating resin treatment of individual reagents was found to decrease copper content of Krebs-Henseleit buffer from 0.32 to 0.17 microM. Using salicylate as a probe for .OH formation, it was determined that considerable amounts of this radical are formed when 0.25 mM ascorbate is added to the buffer indicating significant metal-catalysed autoxidation. Isolated rat hearts, perfused with non-chelexed Krebs-Henseleit buffer plus 0.25 mM ascorbate for 60 min, sustained moderate injury with developed systolic pressure, +dP/dtmax and -dP/dtmax decreased by 30 to 35% by the end of experiment. Hearts perfused with chelating resin-treated Krebs-Henseleit buffer sustained no significant injury within the same time frame. Furthermore, it was observed that hearts perfused with non-chelexed Krebs-Henseleit buffer accumulate significant amounts of copper depending on the amount of contamination and length of perfusion. Significant effects on post-ischemic end diastolic pressure were observed in hearts perfused with a Krebs-Henseleit buffer subsequently found to be contaminated with high levels of copper. These results clearly demonstrate that adventitious redox-active transition metals may be a confounding factor in experimental results. Further, it is recommended that all perfusion media be routinely examined for adventitious metals and treated if deemed necessary.
S R Powell; R A Wapnir
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of molecular and cellular cardiology     Volume:  26     ISSN:  0022-2828     ISO Abbreviation:  J. Mol. Cell. Cardiol.     Publication Date:  1994 Jun 
Date Detail:
Created Date:  1994-10-18     Completed Date:  1994-10-18     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0262322     Medline TA:  J Mol Cell Cardiol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  769-78     Citation Subset:  IM    
Department of Surgery, North Shore University Hospital, Manhasset, NY 11030.
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MeSH Terms
Ascorbic Acid
Chelating Agents
Copper / analysis*,  chemistry
Drug Contamination
Free Radicals
Heart / drug effects*,  physiology
Indicators and Reagents / standards
Iron / analysis*,  chemistry
Perfusion / methods*,  standards
Rats, Sprague-Dawley
Grant Support
Reg. No./Substance:
0/Buffers; 0/Chelating Agents; 0/Free Radicals; 0/Indicators and Reagents; 50-81-7/Ascorbic Acid; 7439-89-6/Iron; 7440-50-8/Copper
Comment In:
J Mol Cell Cardiol. 1995 Sep;27(9):2075-7, 2079-80   [PMID:  8523467 ]

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