J. W. George lvany and E. P. Heimer' Teacher's College, Columbio University New York, New York 10027
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Quick Paaer Chromatography of Monosaccharides
Recently, the application of rapid paper chromatographic systems to amino acids particularly amenable to demonstrations in the teaching laboratory has been described.= Essentially this involved the use of buffered acetonitrile systems not requiring pre-equilibration and permitting complete development in periods less than 1 hr. Because of the great convenience and substantial senaration canacities of these svstems.3 we have now extended applickion to the resoluti"on of monosaccharides. Because of the im~ortanceof the subiect in the beginning biochemistry laboratory, i t is hoped that the convenience and rapidity of the procedure will find acceptance. References to equipment, manuals, and texts are to be found in the preceding r e p ~ r t . ~ Experimental The most satisfactory combination was the system composed of 60 parts acetonitrile: 40 parts 0.1 M ammonium acetate pH 7. Satisfactory resolution of representative sugars can he achieved in less than 1hr. Five microliters of a 3% solution of each sugar was spotted on Whatman 3 MM paper (20 x 20 cm) and develRG Values of
Some Sugars Using 60 Parts Acetonitrile to 40 Parts 0.1 M Ammonium Acetate pH 7 . Cnmnoond
D-Mannital D-Glueosamine HCl D-GalactosamineHCI D-Maltose D-Glucuronic Acid
Ro
Comparison of two paper chromatographic systems for separating monosaccharides. (1) D-glucuronolactone; (2) D-2-deoxyglucose; (3) Dglucose: (4) 0-glucosamine HCI; (5) D-giucuranic acid: (6) D-glucase6-phosphate.
oped in the ascending direction. After the solvent front reached a height of approximately 15 cm from the spotting line, the paper was removed and air dried. The hands were visualized with alkaline silver oxide reagenL4 Results and Discussion It will be seen from the table that a number of representative monosaccharides are well resolved by the described method. The time required for any separation never exceeds a classroom period. Comparison of the procedure with a widely used system5 is demonstrated in the figure. Derivatives of glucose are neatly resolved and the relationship of suhstituent group to migration pattern demonstrated. It will, of course, he realized that differences in RG of at least 0.2 are needed to effect resolution during the time specified. 'Address all correspondence to this author at Chemical ReDepartment, Hoffmsnn-LaRoehe Inc., Nutley, New Jersey
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Heimer, E. P., J. CHEM. EDUC., 49,547 (1972).
3 Gabriel, T. F., J. of
Distance the band has travelled Rc = Distance glucose has travelled
562
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Journal of Chemical Education
Chrom., 36,518 (1968).
Borders, C. L. Jr., J. CHEM. EDUC., 49,437 (1972). 5 Rendina, G., "Experimental Methods in Modem Biochemistry," W. B. Saunders Co., 1971, p. 144. 4
