By Robert Harmon (Eds.)
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Extra resources for Chemistry and Biology of Nucleosides and Nucleotides
This investigation was supported by the Division of Cancer Treatment, National Cancer Institute, National Institutes of Health, Department of Health, Education, and Welfare, Contract No l-CM-43762. 37 Copyright © 1978 by Academic Press, Inc. All rights of reproduction in any form reserved. ISBN 0-12-326140-6 John A. Montgomery et al. 38 I. INTRODUCTION Chemically bonded stationary phases for use in high-pressure liq uid chromatography (HPLC) were first introduced in 1970- These chemically bonded phases are liquids or solids chemically attached to the support material, usually silica gel, and oriented like bristles on the column of support (1,2).
In typical runs about 250 mg of a mixture can be separated in 45 to 90 min. In the exam ple shown, the following solvent program is used: start at 25% acetonitrile to elute quickly any excess of the dinucleotide block and its pyrophosphate. The tritylated material elutes after a linear gradient (30 min) from 25 to 38% of acetonitrile is applied. Before reusing the column, a step gradient to 58% acetoni trile is applied to wash off tightly bound impurities. The small peak appearing at about 43 min contains the starting hexanucleotide.
D. Stevens, Can. J. Chem. 44, 249 (1966); Y. H. Pau, R. K. Robins, and L. B. Townsend, J. Heterocycl. Chem. 4, 246 (1967). Davoll and B. A. Lowry, J. Am. Chem. Soc. 73, 1650 (1951). 9. A. Montgomery and H. J. Thomas, J. Med. Chem. 15, 305 10. (1972).
Chemistry and Biology of Nucleosides and Nucleotides by Robert Harmon (Eds.)