Open AccessIdentification of Nucleosides in Hydrolysates of Transfer RNA by High‐Resolution Mass Spectrometry
Author(s) -
PANG Henrianna,
SMITH David L.,
CRAIN Pamela F.,
YAMAIZUMI Kazuko,
NISHIMURA Susumu,
McCLOSKEY James A.
Publication year - 1982
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1982.tb06894.x
Subject(s) - mass spectrometry , chemistry , chromatography , nucleoside , mass spectrum , resolution (logic) , vaporization , hydrolysate , transfer rna , analytical chemistry (journal) , nucleic acid , mass , trimethylsilyl , hydrolysis , rna , organic chemistry , stereochemistry , biochemistry , computer science , artificial intelligence , gene
High‐resolution mass spectrometry has been investigated as a technique for identification of modified nucleosides in unfractionated hydrolysates of transfer RNA. The method is based on recognition of predetermined sets of exact mass values which are characteristic of individual nucleoside components. Mass spectra are photographically recorded in a non‐time‐resolved fashion, so that differing rates of nucleoside vaporization are of no consequence. Experimental parameters of sensitivity, spectrometer resolution and rates of vaporization were studied. Under typical conditions, 1–4 μg of tRNA are hydrolyzed, converted to volatile trimethylsilyl derivatives, and mass spectra of the resulting mixture recorded during 3 min at resolution 20000; 75–100% of the minor nucleosides are usually identified in a single recording. The method is complementary to conventional methods of identification which rely on chromatographic mobilities, and can in principle be generally applied to recognition of biologically or chemically modified bases in nucleic acid.