Measurement of deoxyguanosine/thymidine ratios in complex mixtures by high-performance liquid chromatography for determination of the mole percentage guanine + cytosine of DNA

J Chromatogr. 1989 Oct 6;479(2):297-306. doi: 10.1016/s0021-9673(01)83344-6.

Abstract

The ratio of deoxyguanosine and thymidine can be determined in a complex mixture containing the major ribonucleosides and deoxynucleosides, the minor deoxynucleosides, and the nucleotide monophosphates by high-performance liquid chromatography. The isocratic procedure utilizes a C18 column and a solvent of methanol-triethylamine phosphate (pH 5.1). A single analysis requires 15 min. Within the range of 0.5-1.5 micrograms of total deoxynucleosides per sample, the determination is very precise and the relative standard deviation is about 0.1%. From the deoxyguanosine/thymidine ratio, a precise determination of the mole percentage guanine + cytosine of double-stranded DNA is calculated.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Archaea / metabolism
  • Chromatography, High Pressure Liquid
  • Cytosine / analysis*
  • DNA / analysis*
  • DNA, Bacterial / analysis
  • Deoxyguanosine / analysis*
  • Guanine / analysis*
  • Hydrolysis
  • Spectrophotometry, Ultraviolet
  • Thymidine / analysis*

Substances

  • DNA, Bacterial
  • Guanine
  • Cytosine
  • DNA
  • Deoxyguanosine
  • Thymidine