Please use this identifier to cite or link to this item: http://dlib.scu.ac.ir/handle/2027.42/24889
Title: A method to reduce interference by sucrose in the detection of thiobarbituric acid-reactive substances
Authors: Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA;Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA
Publisher: Elsevier
Description: A thiobarbituric acid (TBA) reaction for measuring lipid peroxidation products was evaluated for interference by several ingredients commonly used in solutions to prepare or analyze tissue homogenates or subcellular organelles. These included sucrose (up to 100 m final concentration in the assay medium), Tris-maleate (up to 40 m), imidazole (up to 20 m), inorganic phosphate (up to 10 m), and 4-morpholinepropanesulfonic acid (up to 20 m). When the samples were heated at 95[deg]C as recommended in some procedures, only sucrose significantly affected color development. Sucrose concentrations as low as 10 m significantly increased absorbance at 532 nm of aqueous tetramethoxypropane (TMP) standards, and so the assay could not be applied reliably to tissue samples prepared in sucrose. Sucrose interference was only partially reduced by subsequent organic extraction (n-butanol plus pyridine), with measured absorbances remaining significantly greater (50-100%) than sucrose-free controls at sucrose concentrations of 20 m or more. Modifying the assay to include sucrose in blanks and TMP standards failed to adequately correct for interference when the absorbance of unextracted (aqueous) solutions was measured. Further modification by adding sucrose to blanks and TMP standards, followed by butanolpyridine extraction, gave standard curves that were linear, through the origin, and had slopes equivalent to those of sucrose-free standards. This modification enabled almost complete recovery (average 2% error) of known amounts of TMP added to aliquots of tissue homogenates containing amounts of sucrose that otherwise significantly interfered. Also, with the modified method the content of TBA-reactive substances in tissues homogenized in sucrose was found to be not significantly different from that measured in tissues homogenized in a noninterfering substance, KCl.
URI: https://deepblue.lib.umich.edu/handle/2027.42/24889
More Information: Shlafer, Marshal, Shepard, Beverly M. (1984/03)."A method to reduce interference by sucrose in the detection of thiobarbituric acid-reactive substances." Analytical Biochemistry 137(2): 269-276. <http://hdl.handle.net/2027.42/24889>
http://www.sciencedirect.com/science/article/B6W9V-4DYM9JV-12K/2/2bb7d3ac0e44a4ca167db5dbf6047993
http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6731812&dopt=citation
http://hdl.handle.net/2027.42/24889
6731812
http://dx.doi.org/10.1016/0003-2697(84)90084-8
Analytical Biochemistry
Appears in Collections:Research Collections

Files in This Item:
Click on the URI links for accessing contents.
Title: A method to reduce interference by sucrose in the detection of thiobarbituric acid-reactive substances
Authors: Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA;Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA
Publisher: Elsevier
Description: A thiobarbituric acid (TBA) reaction for measuring lipid peroxidation products was evaluated for interference by several ingredients commonly used in solutions to prepare or analyze tissue homogenates or subcellular organelles. These included sucrose (up to 100 m final concentration in the assay medium), Tris-maleate (up to 40 m), imidazole (up to 20 m), inorganic phosphate (up to 10 m), and 4-morpholinepropanesulfonic acid (up to 20 m). When the samples were heated at 95[deg]C as recommended in some procedures, only sucrose significantly affected color development. Sucrose concentrations as low as 10 m significantly increased absorbance at 532 nm of aqueous tetramethoxypropane (TMP) standards, and so the assay could not be applied reliably to tissue samples prepared in sucrose. Sucrose interference was only partially reduced by subsequent organic extraction (n-butanol plus pyridine), with measured absorbances remaining significantly greater (50-100%) than sucrose-free controls at sucrose concentrations of 20 m or more. Modifying the assay to include sucrose in blanks and TMP standards failed to adequately correct for interference when the absorbance of unextracted (aqueous) solutions was measured. Further modification by adding sucrose to blanks and TMP standards, followed by butanolpyridine extraction, gave standard curves that were linear, through the origin, and had slopes equivalent to those of sucrose-free standards. This modification enabled almost complete recovery (average 2% error) of known amounts of TMP added to aliquots of tissue homogenates containing amounts of sucrose that otherwise significantly interfered. Also, with the modified method the content of TBA-reactive substances in tissues homogenized in sucrose was found to be not significantly different from that measured in tissues homogenized in a noninterfering substance, KCl.
URI: https://deepblue.lib.umich.edu/handle/2027.42/24889
More Information: Shlafer, Marshal, Shepard, Beverly M. (1984/03)."A method to reduce interference by sucrose in the detection of thiobarbituric acid-reactive substances." Analytical Biochemistry 137(2): 269-276. <http://hdl.handle.net/2027.42/24889>
http://www.sciencedirect.com/science/article/B6W9V-4DYM9JV-12K/2/2bb7d3ac0e44a4ca167db5dbf6047993
http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6731812&dopt=citation
http://hdl.handle.net/2027.42/24889
6731812
http://dx.doi.org/10.1016/0003-2697(84)90084-8
Analytical Biochemistry
Appears in Collections:Research Collections

Files in This Item:
Click on the URI links for accessing contents.
Title: A method to reduce interference by sucrose in the detection of thiobarbituric acid-reactive substances
Authors: Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA;Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA
Publisher: Elsevier
Description: A thiobarbituric acid (TBA) reaction for measuring lipid peroxidation products was evaluated for interference by several ingredients commonly used in solutions to prepare or analyze tissue homogenates or subcellular organelles. These included sucrose (up to 100 m final concentration in the assay medium), Tris-maleate (up to 40 m), imidazole (up to 20 m), inorganic phosphate (up to 10 m), and 4-morpholinepropanesulfonic acid (up to 20 m). When the samples were heated at 95[deg]C as recommended in some procedures, only sucrose significantly affected color development. Sucrose concentrations as low as 10 m significantly increased absorbance at 532 nm of aqueous tetramethoxypropane (TMP) standards, and so the assay could not be applied reliably to tissue samples prepared in sucrose. Sucrose interference was only partially reduced by subsequent organic extraction (n-butanol plus pyridine), with measured absorbances remaining significantly greater (50-100%) than sucrose-free controls at sucrose concentrations of 20 m or more. Modifying the assay to include sucrose in blanks and TMP standards failed to adequately correct for interference when the absorbance of unextracted (aqueous) solutions was measured. Further modification by adding sucrose to blanks and TMP standards, followed by butanolpyridine extraction, gave standard curves that were linear, through the origin, and had slopes equivalent to those of sucrose-free standards. This modification enabled almost complete recovery (average 2% error) of known amounts of TMP added to aliquots of tissue homogenates containing amounts of sucrose that otherwise significantly interfered. Also, with the modified method the content of TBA-reactive substances in tissues homogenized in sucrose was found to be not significantly different from that measured in tissues homogenized in a noninterfering substance, KCl.
URI: https://deepblue.lib.umich.edu/handle/2027.42/24889
More Information: Shlafer, Marshal, Shepard, Beverly M. (1984/03)."A method to reduce interference by sucrose in the detection of thiobarbituric acid-reactive substances." Analytical Biochemistry 137(2): 269-276. <http://hdl.handle.net/2027.42/24889>
http://www.sciencedirect.com/science/article/B6W9V-4DYM9JV-12K/2/2bb7d3ac0e44a4ca167db5dbf6047993
http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6731812&dopt=citation
http://hdl.handle.net/2027.42/24889
6731812
http://dx.doi.org/10.1016/0003-2697(84)90084-8
Analytical Biochemistry
Appears in Collections:Research Collections

Files in This Item:
Click on the URI links for accessing contents.