Measurement of Deoxyguanosine/thymidine Ratios in Complex Mixtures by High-performance Liquid Chromatography for Determination of the Mole Percentage Guanine + Cytosine of DNA

Overview

Journal J Chromatogr

Date 1989 Oct 6

PMID 2509507

Citations 24

Authors

M Mesbah

W B Whitman

Affiliations

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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.

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