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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1980-8-25
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pubmed:abstractText |
High-performance liquid chromatographic (HPLC) assay for determining tissue pools of uridine, deoxyuridine, cytidine, deoxycytidine, and thymidine mono-, di-, and triphospates is presented. The method utilizes anion-exchange and, after conversion of nucleotides to nucleosides by acid phosphatase, reversed-phase chromatography on a preparative column with UV detection at 254 and 280 nm. The yield of this procedure is 80 +/- 2% with a sensitivity limit of 100 pmole nucleotide per sample. A sensitivity of 10 pmole can be achieved for each compound by rechromatographing appropriate nucleoside fractions on analytical columns. The recovery, including this step, is 66 +/- 7%. The assay is reproducible and highly selective, with a lower sensitivity limit of approximately 0.1 muM using 150--250 mg (wet weight) tissue samples. Nucleotide pools have been determined in Balb/c mouse liver and in mouse lymphoma (S-49) cell culture, the latter with and without addition of 5-fluorouracil (5-FUra) to the medium. Data obtained with this assay are similar to those using alternative methodologies. Observed depletion of dTXP pools and expansion of dUMP and dCXP pools after 5-FUra treatment are in agreement with published observations. Pools of dUDP and dUTP were not detectable (less than 10 pmole/10(8) cells) in any tissue sample. These data illustrate the utility of the present method in studying actions of pyrimidine antimetabolites.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0021-9673
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
25
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pubmed:volume |
188
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
149-58
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:7380929-Animals,
pubmed-meshheading:7380929-Cells, Cultured,
pubmed-meshheading:7380929-Chromatography, High Pressure Liquid,
pubmed-meshheading:7380929-Liver,
pubmed-meshheading:7380929-Mice,
pubmed-meshheading:7380929-Mice, Inbred BALB C,
pubmed-meshheading:7380929-Pyrimidine Nucleosides,
pubmed-meshheading:7380929-Thymine Nucleotides
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pubmed:year |
1980
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pubmed:articleTitle |
Determination of pyrimidine ribotide and deoxyribotide pools in cultured cells and mouse liver by high-performance liquid chromatography.
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.
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