Determination of guanine by high-performance liquid chromatography with electrochemical detection: application to DNA and RNA assays.
A highly sensitive assay for guanine was developed using high-performance liquid chromatography with electrochemical detection (ECD). Guanine was susceptible to the electrochemical oxidation, and ECD response was proportional to the amount of guanine in the range 0.25-4 pmol of guanine. The ECD of guanine was applicable to the analysis of nucleic acids. DNA and RNA were hydrolyzed in 0.03 and 3 M HCl, respectively, and guanine liberated from the nucleic acids was separated on a reverse-phase column and determined by ECD. The method allowed detection of 0.2 ng of calf thymus DNA or tRNA. An application of the method is shown for DNA and RNA assays in trichloroacetic acid extracts of rat adrenal and liver.
Nakahara
T
T
Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka, Japan.
Shiraishi
A
A
Hirano
M
M
Matsumoto
T
T
Kuroki
T
T
Tatebayashi
Y
Y
Tsutsumi
T
T
Nishiyama
K
K
Ooboshi
H
H
Nakamura
K
K
D000311:Adrenal Glands / Q000032:analysis; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA / Q000032:analysis*; D004563:Electrochemistry; D006147:Guanine / Q000032:analysis*; D006851:Hydrochloric Acid; D006868:Hydrolysis; D008099:Liver / Q000032:analysis; D008297:Male; D012313:RNA / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D014238:Trichloroacetic Acid
D014238:Trichloroacetic Acid; D006147:Guanine; D012313:RNA; D004247:DNA; D006851:Hydrochloric Acid