Nucleoside derivatives, high-performance

In a second procedure, poly(ADP-ribose) was first separated from the bulk of the nucleic acids and proteins by dihydroxyboryl-Sepharose affinity chromatography 147,190). The isolated polymer was treated with snake venom phosphodiesterase and bacterial alkaline phosphatase to yield the nucleoside 2, l"-ribosyladenosine from internal residues. This product was then treated with chloroacetaldehyde to produce the fluorescent derivative, l,iSr -ethenoribosyladenosine, which was then separated from other derivatized residues by reversed-phase high performance liquid chromatography picomole amounts were quantified by fluorescence detection. This procedure facilitates the accurate determination of minute quantities of endogenous poly(ADP-ribose) (102, 190). Niedergang et al. (147) have also utilized a fluorimetric assay for determination of the enzymatic digestion products of the polymer, ADP-ribose, or iso-ADP-ribose. 

Unidimensional TLC of nucleic acid derivatives has also been done on silica gel. Issaq et al. (1977) separated mixtures of alkylated guanines, adenines, uracils, and cytosines on silica gel 6OF2S4, in various chloroform-alcohol solvent systems. The addition of 1 ml of ammonium hydroxide to the mobile phase prevented streaking and resulted in nondistorted developed spots. Figueira and Ri-beiro (1983) separated adenine, adenosine, inosine, hypoxanthine, 2-AMP, 5-AMP, ADP, ATP, cAMP, and dibutyryl cAMP also on silica gel 6OF254, and Sleckman et al. (1985) used conventional and high-performance silica gel to separate four representative nucleotides and their corresponding nucleosides. Detailed Experiment 2 of this chapter describes a unidimensional separation of nucleotides and nucleosides on silica gel. 

Nucleic acid base and nucleoside derivatives were bonded to 3-aminopropyl silanized silica (APS-silica) and silica gel. These resins were useful as the columns of high performance liquid chromatography (HPLC) for the selective separation of oligoethyleneimine derivatives having pendant thymine or adenine bases. These column systems were also found to be applicable to the separation of nucleosides, nucleotide, and oligonucleotides. 

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