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Clinical Chemistry 18: 810-813, 1972;
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Clinical Chemistry, Vol 18, 810-813, Copyright © 1972 by the American Association for Clinical Chemistry

Gas-Liquid Chromatographic Analysis for Purine and Pyrimidine Bases in Hydrolysates of Nucleic Acid

Duane B. Lakings 1 and Charles W. Gehrke 1

1 Experiment Station Chemical Laboratories, University of Missouri, Columbia, Mo. 65201.

In this fast, simple method for analysis of microgram amounts of purine and pyrimidine bases in nucleic acid hydrolysates, RNA and DNA are optimally hydrolyzed in equal volumes of trifluoroacetic acid and formic acid, heated in a closed tube at 200°C for 1.5 h (for RNA) or at 150°C for 2.0 h (for DNA). The trifluoroacetic acid and formic acid are evaporated from the hydrolysates on a hot plate at 60°C, under a stream of pure nitrogen. The resulting nucleic acid bases are then silylated with a mixture of equal volumes of bis(trimethylsilyl)trifluoroacetamide and acetonitrile at 150°C for 15 min, and analyzed by gas-liquid chromatography on a column of SE-30 on Supelcoport.


Key Words: anion-exchange cleanup • elution • internal standard • relative molar response • yeast- and TMV-RNA • salmon sperm DNA • silylation






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Copyright © 1972 by the American Association for Clinical Chemistry.