Separation of Transfer Ribonucleic Acids by Reverse Phase Chromatography

A New Chromatographic System for Increased Resolution of Transfer Ribonucleic Acids*

“…The position of the 14 tRNAs was determined by amino acid acceptance assay and 12 were found to be heterogeneous.3 For example, at pH 7.0, five leucine tRNAs were found, while at pH 4.5, three peaks each were detected for alanine, arginine, phenylalanine, and methionine and at least two peaks were present for aspartic acid, histidine, isoleucine, lysine, proline, tyrosine, and valine tRNAs. A greater degree of heterogeneity for many tRNAs was observed with this Freon solvent reversed-phase chromatographic system than was previously observed with a similar system using isopentyl acetate and dimethyldilaurylammonium chloride (Kelmers et al, 1965).…”

“…A new solvent system, employing quaternary ammonium extractants in a Freon diluent (Khym, 1966), was modified by selection of a high-boiling Freon1 and has been tested as a reversed-phase column chromatographic system. This system is in principle no different from the reversed-phase system described earlier (Kelmers et al, 1965). A new solvent, new ion exchanger, and exploration of gradient possibilities yielded superior resolution and a more even distribution of tRNAs throughout the chromatogram.…”

“…A reversed-phase chromatographic system employing quaternary amines in an organic diluent was devised (Kelmers et al, 1965) and applied to the preparation of highly purified Escherichia coli B phenylalanine tRNA (Kelmers, 1966a) and a leucine tRNA (Kelmers, 1966b). However, this system was not equally useful for other tRNAs, as many were nearly superimposed in the early portions of the chromatogram.…”

Analysis of Isoaccepting Transfer Ribonucleic Acid Species of Bacillus subtilis : Chromatographic Differences Between Transfer Ribonucleic Acids from Spores and Cells in Exponential Growth

“…This is a much studied species in E. coli tRNA because of the changes which occur in isoaccepting species of leucyl-tRNA during infection of E. coli by phage T2 (12). Five isoaccepting leucine tRNA species have been observed in E' coli (14). No changes in leu-tRNA during sporulation in Bacillus subtilis have been reported.…”

Hydrophobicity of polymer based anion-exchange columns for ion chromatography

“…There are numerous mixed-mode stationary phases available, but for the majority of them, hydrophobic interactions are considered to be dominant for retention/separation in HPLC, while ion-exchange interactions play a secondary role and are responsible only for tuning of separation selectivity towards specific ionisable analytes [ 17 , 20 , 21 , 22 ]. As the majority of bio- and pharmaceutical molecules separated by reversed phase HPLC (RP HPLC) are charged, mixed-mode stationary phases may be extremely useful as it was highlighted by Kelmers [ 23 ]. However, a lack of understanding of secondary electrostatic interactions for this type of stationary phases makes their practical use more difficult.…”