ABSTRACT
CHCl3/Tol/HForm or n-BuOH/NH3 or CHCl3
Aldehydes Silica gel G EtOAc/Hex 30 Aldehydes, 2,4dinitrophenylhydrazones
Alumina
Alumina IB Silica gel Silica gel IB
C6H6 or CHCl3 or Et2O or C6H6/Hex
MeCl or Tol/THF (4:1) Hex/EtOAc (4:1 or 3:2) MeCl or Tol/THF (4:1)
32 3 32
Alkaloids Alumina
Alumina
Cellulose (impregnated with formamide)
Paper (S&S 2043b) Paper electrophoresis
Silica gel
i-BuOH/AcOH or i-BuOH/NH3 or i-PrOH/AcOH
CHCl3 or EtOH or (CH2)6/CHCl3(3:7) C6H6/n-C7H16/CHCl3/Et2NH (6:5:1:0.02) n-BuOH/HCl(25%)/w (100:26:39) i-BuOH/AcOH or i-BuOH/NH3 or i-PrOH/HOAc
C6H6/EtOH (9:1) or CHCl3/Ac/Et2NH (5:4:1)
34 35
Amides Kieselguhr (adipic acid impregnated)
Silica gel
i-Pr2O/Petet/CCl4/HForm/w
i-Pr2O/Petet/CCl4/HForm/w
36 Amines Alumina
Alumina G Keiselguhr G Silica gel Silica gel (aromatic only)
Ac/n-C7H16 (1:1) i-BuAc or i-BuAc/AcOH Ac/w (99:1) EtOH (95%)/NH3 (25%) (4:1)
3 37 3 3 38
Amino acids Alumina Cellulose Cellulose (two-dimensional)
Silica gel
n-BuOH/AcOH/w (3:1:1) or Py/w n-BuOH/AcOH/w (4:1:1) n-BuOH/Ac/NH3/w (10:10:5:2) followed by i-PrOH/HForm/w (20:1:5)
n-BuOH/AcOH/w (3 or 4:1:1) or PhOH/w (3:1) or n-PrOH/NH3 (34%) (2:1)
3 3 3
Aminochromes Whatman 1 (acid washed) w or AcOH/w or MeOH/w or EtOH/w or n-BuOH/AcOH/w or i-PrOH/w
26, 39
Barbiturates Silica gel CHCl3/n-BuOH/NH3 (25%) (14:8:1) 3 Benzophenones, hydroxy
Alumina or cellulose or Kieselguhr (impregnated with adipic acid triethylene glycol polyester) or silica gel
HForm/m-X 40
Bile acids Silica gel C6H6/Et2O (4:1) or Et2O/AcOH (99.6:0.4) or CHCl3/MeOH (9:1) 41
Caffeine Chromatography paper n-BuOH/NH3 or n-BuOH/HForm 42 Carboxylic acids Keiselguhr/polyethylene glycol
Polyamide powder
Silica gel G Silica gel (CaSO4 impregnated) Silica gel/polyethylene glycol M-1000
i-Bu2O/HForm/w (90:7:3) i-Pr2O/Petet/CCl4/HForm/w (50:20:20:8:1) or MeCN/EtOAc/HForm or BuForm/EtOAc/HForm
EtOH/NH3/THF n-PrOH/NH3 or EtOH/CHCl3/NH3 i-Pr2O/HForm/w (90:7:3)
43 43
30 44 43, 45, 46
Family Stationary Phase Mobile Phase Reference Carboxylic acids, unsaturated
Silica gel CHCl3/MeOH 47
Catecholamines Alumina Boric acid gel (neutral pH) Kieselguhr Phenylboronate Phosphocellulose
HCl (0.025 N) Dilute acids Ph buffer (pH = 6.2)/EDTA
48 48 49 50 51
Catecholamines, dansyl derivatives
Alumina (two-dimensional) Amberlite IRC50
C6H6/EtOAc (60:40) or CHCl3/EtOH/Tol (90:6.5:3.5)
52 Catecholamines, o-methyl derivatives
Silica gel (sodium borate impregnated)
Corticosteroids Silica gel EtOH(5%)/MeCl or EtOH/CHCl3 54, 55 Coumarins Polyamide
Silica gel G Silica gel G (impregnated with NaAc)
Silicic acid (starch bound)
MeOH/w (4:1 or 3:2) Petet/EtOAc (2:1) Tol/EtFor/HForm (5:4:1) EtOAc/Skellysolve B
3 3 3
3 Dicarboxylic acids Kieselguhr/polyethylene glycol
Polyamide powder
Polyamide Woelm DC powder
Silica gel Silica gel G
i-Pr2O/HForm/w (90:7:3) i-Pr2O/Petet/CCl4/HForm/w (50:20:20:8:1) or MeCN/EtOAc/HForm (9:1:1) or BuFor/EtOAc/HForm (9:1:1)
MeCN/PrFor/PrAc/HForm (45:45:10:10) or iPr2O/Petet/CCl4/HForm/w (50:20:20:8:1) or nAmOH/CCl4/HFor (3:2:1)
i-Pr2O/HForm/w (90:7:3) EtOH/NH3/THF
43 43
43 30
Diols (see Alcohols, polyhydric)
Disulfides Alumina Hex 57 Disulfides, 3,5dinitro-benzoates
Whatman 3 (impregnated with 10% paraffin oil in (CH2)6)
DMF/MeOH/w or Foram/MeOH/w 58
Flavinoids Paper
Polyamide Silica gel
Silica gel (impregnated with NaAc)
Silicic acid (starch bound)
n-BuOH/AcOH/w or EtOAc/w or AcOH/w or C6H6/AcOH/w
MeOH/H2O C6H6/Py/AcOH (36:9:5) Petet/EtOAc (2:1) Tol/EtForm/HFor (5:4:1) EtOAc/Skellysolve B
3 59
3 Glycerides Silica gel G
Silica gel G (impregnated with silver nitrate)
CHCl3/C6H6 (7:3) CHCl3/AcOH (99.5:0.5)
3 3
Glycolipids Silica gel G n-PrOH/NH3 (12%) (4:1) 3 Glycols, polyethylene Paper
Silica gel
n-PrOH/EtOAc/w (7:1:2) or n-BuOH/AcOH/w (4:1:5) or t-AmOH/n-PrOH/w (8:2:3) or EtOAc/AcOH/w (9:2:2)
Ac or n-BuOH/AcOH/w
61 Hydroxamates Silica gel i-Pr2O or i-Pr2O/EtOAc (1:4) or
i-Pr2O/i-C8H18 62
Hydroxamic acids Kieselguhr G (impregnated with diethylene glycol or triethylene glycol adipate polyesters)
i-Pr2O/Petet/CCl4/HForm/w (50:20:20:8:1) 98
Indoles Acetylated (ascending) Cellulose (thin-layer)
CHCl3/MeOH/w (10:10:6) w or HCl (0.005 N) or n-BuOH/AcOH/w (12:3:5) or C6H6/AcOH/w (125:72:3)
63 64
α-Ketoacids Silica gel (CaSO4 impregnated) EtOH/CHCl3/NH3 44
Family Stationary Phase Mobile Phase Reference Ketones, 2,4-dinitrophenyl hydrazones
Alumina IB Silica gel IB
MeCl or Tol/THF (4:1) MeCl or Tol/THF (4:1)
32 32
Lactams Silica gel i-Pr2O or i-Pr2O/EtOAc (1:4) or i-Bu2O/i-C8H18
Lactones Silica gel i-Pr2O or i-Pr2O/EtOAc (1:4) or i-Bu2O/i-C8C18
Lipids Alumina Silica gel G Silicic acid
Petet/Et2O (95:5) Petet/Et2O/AcOH (90:10:1) CHCl3/MeOH/w (80:25:3)
3 3 3
Mercaptans (see Thiols)
Nitrosamines Silica gel Kieselgel
Hex/Et2O/MeCl MeCl/Hex/Et2O (2:3:4) (aliphatic, aromatic); MeCl/Hex/Et2O (5:7:10) (cyclic)
65 66
Nucleotides Cellulose
Cellulose (on DEAE) AmSO4 (sat’d)/NaAc(1 M)/i-PrOH (80:18:2) HCl (aq)
3 Oximes Silica gel G C6H6/EtOAc or C6H6/MeOH (abs) 67 Peroxides Silicone filter paper w/EtOH/CHCl3 68 Phenols Alumina
Alumina/AcOH Silica gel A
Silica gel G Silica gel/oxalic acid Silica gel/potassium carbonate
Et2O C6H6 CHCl3/AcOH (5:1) or CHCl3/Ac/AcOH (10:2:1) or C6H6/AcOH (5:1) or Petet (80°)/CCl4/AcOH (4:6:1) or CHCl3/Ac/Et2NH (4:2:0.2)
C6H6/Diox/AcOH (90:25:4) C6H6 MeCl/EtOAc/Et2NH (92:5:3 or 93:5:2)
3 3 69
3 70 70
Phosphates, esters Alumina
Kieselgel
Hex/C6H6/MeOH (2:1:1) or Hex/MeOH/Et2O
Hex/C6H6/MeOH (2:1:1) or Hex/MeOH/Et2O
Phospholipids Silica gel G CHCl3/MeOH/w 3 Polynuclear aromatics
Alumina Alumina Silica gel
CCl4 C6H6/(CH2)6 (15:85) Hex or CH3CHCl2 or C2HCl3 or CCl4
3 72 73, 74
Polypeptides Sephadex G-25 Silica gel G
w or NH3 (0.05 M ) CHCl3/MeOH (9:1) or CHCl3/Ac (9:1)
3 3
Pyridines Whatman 1 (descending) n-BuOH/w or n-BuOH/w/NH3 or Ac or i-BuOH/w or MEK/AcOH/w
Pyridines, quaternary salts (descending)
Whatman 1 Ac/w or AmSO4/Ph buffer (pH = 6.8)/n-PrOH(2%) or n-PrOH
Purines Silica gel Ac/CHCl3/n-BuOH/NH3 (25%) (3:3:4:1) 3
Pyrrole, tricarboxylic acid
Silica gel n-BuOH/EtOH/NH3/w (10:10:1:1) 64
Skatoles, hydroxy Silica gel G i-Pr2O or (ClCH2)2/i-Pr2NH (6:1) 76 Steroids Alumina
Paper
Paper (impregnated with kerosene)
Silica gel G
CHCl3EtOH (96:4) Petet/Tol/MeOH/w or Petet/C6H6/MeOH/w
n-PrOH/w EtOAc/(CH2)6/EtOH(abs) or EtOAc/(CH2)6 or CHCl3/EtOH (abs) or C6H6/EtOH or n-C6H14/EtOAc or EtOAc/n
C6H14/EtOH(abs)/AcOH or EtOAc/n-C6H14/AcOH
3 60
78, 79 80, 81
Family Stationary Phase Mobile Phase Reference Sugars Cellulose
Kieselguhr G (buffered with 0.02 N NaAc)
Silica gel (buffered with H3BO3) Silica gel (impregnated with sodium bisulfite)
Silica gel G Whatman 1 (descending, twodimensional)
n-BuOH/Py/w (6:4:3) or EtOAc/Py/w (2:1:2)
EtOAc/i-PrOH/w
C6H6/AcOH/MeOH (1:1:3) EtOAc/AcOH/MeOH/w (6:1:5:1) or n-PrOH/w (85:15) or i-PrOH/EtOAc/w (7:1:2) or MEK/AcOH/w (6:1:3)
n-PrOH/conc. NH3/w (6:2:1) PhOH or n-BuOH/AcOH
3 3
3 82
Sugars, aldoses Paper
Whatman 1
EtOAc/Py/w (2:1:2) or n-BuOH/AcOH/w (4:1:5) or n-BuOH/EtOH/H2O (5:1:4) or EtOAc/AcOH/w (9:2:2) or EtOAc/AcOH/HForm/w or EtOAc/Py/NaAc (sat’d)
PhOH or n-BuOH/AcOH
82 Sugars, carbamates Silica gel n-BuOH/H3BO3 (0.03 M) (9:1) Sugars, deoxy Whatman 1 PhOH or n-BuOH/AcOH 82 Sugars, ketoses Paper
Whatman 1
EtOAc/Py/w (2:1:2) or n-BuOH/AcOH/w (4:1:5) or
n-BuOH/EtOH/H2O (5:1:4) or EtOAc/AcOH/w (9:2:2) or w/PhOH (pH = 5.5)
PhOH or n-BuOH/AcOH
82 Sulfides Alumina
Alumina Silica gel Silica gel DF-5
Hex CHCl3/MeOH CCl4 or C6H6 Ac/C6H6 or Tol/EtOAc
75 96 83 95
Sulfilimines, pnitrosobenzene sulfonyl
Whatman 4 (impregnated with formamide)
C6H6 or C6H6/(CH2)6 84
Sulfonamides Kieselguhr
Silica gel Silica gel (neutral) Silica gel G
CHCl3/MeOH (9:1) or CHCl3/MeOH/NH3
Et2O or CHCl3/MeOH (10:1) n-BuOH/MeOH/Ac/Et2NH (9:1:1:1) CHCl3/EtOH/n-C7H16
86 87
3 Sulfones Alumina
Silica gel DF-5 Et2O or Hex/Ac (1:1) Ac/C6H6 or Tol/EtOAc
57 95
Sulfones, esters Alumina Et2O or Hex/Ac (1:1) 57 Sulfones, hydroxyethyl
Alumina Hex/w (1:3) 57
Sulfoxides Alumina Alumina Silica gel Silica gel DF-5 Whatman 1
C6H6/Py (20:1) and Diox Ac/CCl4 (1:4) Ac or EtOAc or CHCl3/Et2O Ac/C6H6 or Tol/EtOAc PhOH/w (8:3) or n-BuOH/AcOH/w (9:1:2.5)
88 97 83 95 89
Sulfoxides, hydroxyethyl
Alumina Et2O or Hex/Ac (1:1) or Hex/Et2O (1:3) 57
Terpenes Alumina Silica gel G Silica gel/gypsum Silicic acid (starch bond)
C6H6 or C6H6/Petet or C6H6/EtOH i-Pr2O or i-Pr2O/Ac CHCl3/C6H6 (1:1) n-C6H14/EtOAc (85:15)
3 3 90 3
Thiobarbiturates Paper n-AmOH/n-BuOH/25% NH3(2:2:1) 91
Thiolactones Silica gel i-Pr2O or i-Pr2O/EtOAc (1:4) or i-Bu2O/i-C8H18
Family Stationary Phase Mobile Phase Reference Thiols Alumina
Alumina (activated) Alumina (5% cetane impregnated)
Silica gel
Hex AcOH/MeCN (3:1) AcOH/MeCN (3:1) EtOAc or CHCl3
57 96 96
83 Thiophenes Alumina G
Silica gel Petet (40-60°C) MeOH or C6H6/CHCl3 (9:1)
92 92
Thiophosphate, esters
Petet or C6H6/CHCl3 or Ac or EtOH or EtOAc or MeOH
Ureas Acetylated plates Silica gel
CCl4/EtOAc/EtOH (100:5:2) CCl4/MeCl/EtOAc/HOAc (70:50:15:10)
94 94
Urethanes (see Ureas)
Typical Stationary and Mobile Phase Systems Used in the Separation of Various Inorganic Ions
Stationary Phase Mobile Phase Solvent Ratio Separated Ions Silica gel G Butanol/1.5 N HCl/2,5-
hexanedione 100:20:0.5 Hydrogen sulfide group
Silica gel G Acetone/conc. HCl/2,5hexanedione
100:1:0.5 Ammonium sulfide group
Silica gel G Water sat’d ethyl acetate/tributyl phosphate
100:4 U, Ga, Al
Silica gel G Ethanol/acetic acid 100:1 Alkali metals Silica gel G Acetone/1-butanol/conc.
