Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1980-8-25
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.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0021-9673
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
188
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
149-58
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1980
pubmed:articleTitle
Determination of pyrimidine ribotide and deoxyribotide pools in cultured cells and mouse liver by high-performance liquid chromatography.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S.