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Paper Chromatography of Amino Acids - R_f Values

	Solvent Systems
Amino Acid	1	2	3	4	5	6
ALA alanine	0.58	0.30	0.11	0.09	0.45	0.22
ARG arginine	0.56	0.138	0.03	0.05	0.22	0.07
ASP aspartic acid	0.18	0.106	0.03	0.10	0.27	0.09
CYS cysteine
CYS2 cystine				0.01	0.04	0.11
GLU glutamic acid	0.31	0.12	0.05	0.01	0.37	0.11
GLY glycine	0.39	0.22	0.06	0.05	0.32	0.19
HIS histidine	0.64	0.25	0.03	0.09	0.22	0.22
ILE isoleucine	0.84	0.55	0.36	0.40	0.62	0.47
LEU leucine	0.84	0.55	0.34	0.46	0.64	0.52
LYS lysine	0.48	0.08	0.02	0.02	0.11	0.16
MET methionine				0.05	0.48	0.40
PHE phenylalanine	0.84	0.58	0.29	0.38	0.55	0.58
PRO proline	0.86	0.31	0.14		0.47	0.29
SER serine	0.36	0.23	0.05	0.02	0.32	0.29
THR threonine	0.49	0.30	0.09	0.04	0.45	0.51
TRP tryptophan	0.80	0.66	0.22	0.30	0.35	0.41
TYR tyrosine	0.67	0.64	0.16	0.19	0.41	0.34
VAL valine	0.78	0.44	0.24	0.15	0.63	0.37
ASN asparagine	0.44	0.172	0.02
GLN glutamine	0.60	0.21	0.04

Solvent Systems: (all are odiforous, but the ones in red stink)

1. Phenol saturated with water (pH 5.0 - 5.5) [a]
2. Collidine-lutidine (1:3) saturated with water. [a]
3. n-Butanol-acetic acid (9:1) satuarated with water. [a]
4. n-Butanol satuarted with 2M NH₄OH. [b]
5. Methanol-water-pyridine (20:5:1). [c]
6. t-butanol-methylethyl ketone-water-diethylamine (10:10:5:1). [c]
   (This system will give a strong ninhydrin background, unless the diethylamine is removed. This can be accomplished by spraying the chromatogram with 0.1 M borate buffer, pH 9.3, and drying over a hot plate or in a 100°C oven.)

1. Lederer and Lederer; Chromatography, 2nd Ed. Elsevier, 1957, pp 312 and 328.
2. Block, Durrum and Zweig; A Manual of Paper Chromatography and Paper Electrophoresis, Academic Press (1956) p 108.
3. Redfield; Biochimica et Biophysica Acta 10: 344 (1953).