CHROMATOGRAPHY OF PURINES AND PYRIMIDINES ON STARCH COLUMNS
1949; Elsevier BV; Volume: 179; Issue: 2 Linguagem: Inglês
10.1016/s0021-9258(19)51291-1
ISSN1083-351X
Autores Tópico(s)Protein purification and stability
ResumoVVe have found that it is possible to obtain satisfactory resolution of the six bases, thymine, uracil, cytosine, adenine, guanine, and hypoxanthine, by chromatography on a single column of the type developed by Stein and Moore' for amino acids.On known mixtures recoveries have averaged 100 f 4 per cent.The bases are easily identified by their absorption spectra which are characteristic, and not appreciably changed by passage through the column.The quantitative estimation of purines and pyrimidines by paper chromatography has been described by Vischer and Chargal? and by Hotchkiss;s and Edman et al.' have reported the separation of adenine and guanine on a starch column.The starch columns employed in our work are 30 cm. in height and 0.9 cm. in diameter, and are prepared according to the procedure recommended by Stein and Moore.The solvent is composed of n-propanol and 0.5 N HCl in the proportions of 2:l.A solution containing from 0.20 to 0.35 mg. of each base in 0.5 to 1.0 ml. of solvent is added to the top of the column.The effluent is collected in a regular series of 0.5 ml.fractions6The fractions are evaporated to dryness in groups of 50 to 80 in a vacuum desiccator warmed to about 40' by means of an i&a-red lamp.Each residue is dissolved in 5 ml. of 0.1 N HCl.The extinction coefficients of the solutions are measured in the Beckman spectrophotometer at wave-lengths corresponding to the absorption maximum for each 1
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