702

FRANCIS AND ANDERSEN

Table 2 Rf Values and Colors of Flavonoid Aglycones on Silica Gel, Polyamide, and
Reversed-Phase Layers"
Substituent position15
Pigment, spot color3
Flavones
Flavone, b
5-Hydroxyflavone, br
6-Hydroxyflavone, gr
7-Hydroxyflavone, b
7,8-Dihydroxyflavone, gr-br
Chrysin, br-g
Apigenin, g
Acacetin, gr-g
Apigenin-7', 4 '-dimethyl ether, gr-g
Luteolin, y
Diosmetin, gr
Tangeretin, gr
Flavonols
Galangin, g
Kaempferol, y-g
Kaempferid, b-g
Kaempferol-7, 4 '-dimethyl ether, b-g
Kaempferol-3, 4 '-dimethyl ether, ol
Kaempferol-3,7,4'-trimethyl ether, g
Quercetin, br-o
Tamarixetin, g
Rhamnetin, o
Quercetin-3,7-dimethyl ether, o
Morin, g
Fisetin, o
Robinetin, o
Myricetin, o-br
Quercetagetin, r-br

OH

OCH3

5
6
7
7,8
5,7

5,7,4'
5,7
5

5,7,3',4'
5,7,3'

4'

7,4'
4'

5,6,7,8,4'
3,5,7
3,5,7,4'
3,5,7

4'

3,5
5,7
5

7,4'
3,4'
3,7,4'

3,5,7,3',4'
3,5,7,3'
3,5,3',4'
5,3',4'
3,5,7,2',4'
3,7,3',4'
3,7,3',4',5'
3,5,7,3',4',5'
3,5,6,7,3',4'

System0

4'
7
3,7

0.50
0.63
0.31
0.29
0.22
0.48
0.29
0.42
0.56
0.22
0.23
0.25

0.60
0.74
0.50
0.48
0.43
0.57
0.46
0.54
0.66
0.38
0.43
0.48

0.86
0.88
0.65
0.51
0.32
0.52
0.18
0.45
0.86
0.06
0.41
0.89

0.29
0.22
0.39
0.41
0.57
0.33
0.48
0.30
0.14
0.53
0.46
0.26

0.51
0.32
0.48
0.63
0.43
0.61
0.23
0.31
0.29
0.27
0.17
0.21
0.08
0.13
0.00

0.59
0.48
0.59
0.73
0.55
0.69
0.41
0.48
0.48
0.46
0.29
0.36
0.22
0.32
0.00

0.41
0.14
0.38
0.74
0.61
0.87
0.06
0.19
0.24
0.47
0.02
0.08
0.01
0.02
0.01

0.38
0.57
0.37
0.18
0.35
0.19
0.67
0.53
0.45
0.46
0.73
0.70
0.82
0.76
0.95

"Pigment colors are observed under UV, 366 nm after spraying plates with Naturstoffreagenz A followed
by polyethyleneglycol (PEG-4000). b = blue, br = brown, g = green, gr = gray, o = orange, ol = olive,
r = red, y = yellow.
"See Fig. 2 for structures.
c
Systems: 1 and 2, silica gel (60 F254, 0.25 mm), benzene-ethyl acetate-formic acid (40:10:5) and
toluene-ethyl formate-formic acid (50:40:10); 3, polyamide (DC-Alufolien F254, 0.15 mm), toluenebutan-2-one-methanol (60:25:15); 4, reversed-phase (C-18 F2S4, 0.25 mm) methanol-formic acid-water
(58:10:16).

Replacing a hydroxyl with a methoxyl function usually results in higher Rf values. However, a
methoxyl group at the 3-position cancels the effect of the existing hydrogen bond in the parent
hydroxyl compound, and increased retention is observed.
b. Polyamide Layers. Samples (3 //.L) are applied on a polyamide sheet and developed
over 8.5 cm (25 min) with toluene-2-butanone-methanol (60:25:15) as mobile phase. NP/PEG
4000 is used as detection reagent, and the colors are similar to those produced in the preceding
system. Rf values are given under system 3 in Table 2.