SEPARATION BY PAPER CHROMATOGRAPHY 



263 



TABLE V 

 Rf Values of Pentoses 



Solvent 



Arabinose 



Xylose 



Lyxose 



Ribose 



Rhamnose 



2-Deoxyribose 



" Phenol satd. with water at 20°, with 1% NHj and a few crystals KCN in soln. in bottom of tank; What- 

 man No. 1 paper; Partridge, Biochem. J. 42, 238 (1948). 



*■ Methyl ethyl ketone satd. with water at 20°, with 1% NHj in soln. in bottom of tank; Whatman No. 

 1; Partridge, op. cit. The value for lyxose is calcd. from the data of Crosbie et al., Biochem. J. 54, 287 (1953). 



' n-Butanol 50 vol., ethanol 10 vol., water 40 vol., theupper layer beingused; Whatman No. 1. Rxa values 

 (movement relative to that of 2,3,4,6-tetramethylglucose taken as 1.00) are shown. E. L. Hirst and J. K. N 

 Jones, Discussions Faraday Soc. 7, 271 (1949). 



** To the upper layer resulting from the mixture of 1 vol. pyridine, 1.5 vol. water, and 3 vol. n-butanol, 

 at about 28°, is added 1 vol. pyridine; Schleicher and Schilll No. 597 paper. Chargaff el al., J. Biol. Chem. 

 177,405 (1949). 



be achieved. The bases from 5 ^g- of yeast PNA, each eluted in 0.1 ml. and examined 

 in a cell of 1 cm. path length, would give optical densities in the range 0.25 to 0.5, so 

 that it should be entirely practical to work on this scale without special equipment. 



VI. Chromatography of Nucleic Acid Sugars 



This account would be incomplete without reference to the identification 

 of nucleic acid carbohydrates by paper chromatography. However, since 

 the methods do not differ from those used for other carbohydrates, which 

 have been adequately reviewed,^"®- ''' and since the results with nucleic acid 

 components have been limited in scope, this description will be kept very 

 brief. 



Numerous spray reagents have been described for detection of sugars on 

 paper chromatograms. Ammoniacal silver nitrate, used in the original in- 

 vestigations in this field, ^^ is sensitive but relatively nonspecific and sub- 

 ject to interference by impurities. w-Phenylenediamine^^ forms with a wide 

 range of aldoses and ketoses derivatives which fluoresce in ultraviolet light, 

 and has been used in Chargaff 's laboratory for recognition of nucleic acid 

 sugars. Aniline hydrogen phthalate^"" is a sensitive and convenient reagent 



" S. M. Partridge, Biochem. Soc. Symposia {Cambridge, Engl.) No. 3, 52 (1949). 

 '8 S. M. Partridge, Biochem. J. 42, 238 (1948). 



99 E. Chargaff, C. Levine, and C. Green, J. Biol. Chem. 175, 67 (1948). 

 ""' Aniline, 0.93 g., and phthalic acid, 1.66 g., are dissolved in 100 ml. water-saturated 



butanol. The papers are sprayed and then heated to 105° for 5 min. S. M. Partridge, 



Nature 164,443 (1949). 



