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Senin, 15 Oktober 2012
Analysis of Flavonoids TLC Methods
Analysis of Flavonoids TLC Methods

Antiquity, paper chromatography is widely used for

the analysis of flavonoids, but now more methods of analysis that is simple and
inexpensive Thin Layer Chromatography (TLC / TLC). TLC's advantages are:
• The process of separation of compounds that are relatively short.
• Method detection reagents enough to spray.
• Can be analyzed in several samples at the same time.

TLC is suitable for initial orientation analysis of plant extracts before

continuing to other analysis tools instruments such as HPLC, GC, etc..
Flavonoids have two benzene rings separated by propane and is a
derivative of flavone. In general, water-soluble flavonoid compounds. The more brightly
colored the conjugated compound. In the plants, flavonoid commonly found in the form of
glycosides. The difference in the classification of flavonoids shown by the addition of
oxygen, a heterocyclic ring and hydroxyl groups. These groups include catechins,
leucoanthocyanidin, flavanones, flavanonol, flavones, anthocyanidins, flavonols, Chalcone,
aurone and isoflavones.
There are many kinds of systems solvent / eluent used for the separation of
flavonoids using TLC. One example of the results of the methylation or acetylation of
flavones and flavonols require nonpolar solvents such as chloroform-methanol (15:1). Being
such flavonoid aglycone apigenin, luteolin and quercetin can be separated with chloroform
methanol (96:4) or with the same polarity. In general, the mobile phase TLC for flavonoid
glycosides is ethyl acetate - formic acid - glacial acetic acid - water (100:11:11:26). If the
addition of methyl ethyl ketone (ethyl acetate-methyl ethyl ketone-formic acid-glacial aseta
acid - water (50:30:7:3:10), rutin and vitexin-2''-O-ramnosida be separated.
With regard to detection, flavonoids spot on TLC plates produce a yellow-
brown spots white background when reacted with iodine vapor.
Flavonoids may appear as dark spots on a green background fluoresce
when observed in UV light at 254 nm UV-plates containing fluorescent
indicator (such as silica gel F254). If under 365 nm UV light, spot colors depending on
the structure of flavonoids, can be yellow, green or blue fluorescent. It would be more clear
and intense after being sprayed with the reagent.
Colors can be observed at 365 nm UV light are as follows:

• Quercetin, myricetin, and 3 & 7-O-glycosides: orange-yellow

• kaempferol, isorhamnetin, and 3 & 7-O-glycosides: yellow-green

• Luteolin and 7-O-glycosides: orange

• Apigenin and 7-O-glycosides: yellow-green

Further details regarding the use of reagents natural material products can
be seen diartikel Brasseur and Angenot, 1986, p 351.
Ferric chloride in water or ethanol is generally apparition spot on analysis
of phenolic compounds will provide a blue-black color on the detection of
flavonoids. Similarly, Fast Blue Salt B form a blue or blue-purple.
Here is a list of TLC eluent for the separation of flavonoids on silica gel
stationary phase:
Sampel Eluen
Flavonoid aglycon EtOAc–Isopropanol–H2O, 100:17:13
EtOAc– Chloroform, 60:40
Chloroform–MeOH, 96:4
Toluene– Chloroform –MeCOMe, 8:5:7
Toluene–HCOOEt–HCOOH, 5:4:1
Toluene–EtOAc–HCOOH, 10:4:1
Toluene–EtOAc–HCOOH, 58:33:9
Toluene–EtCOMe–HCOOH, 18:5:1
Toluene–dioxane–HOAc, 90:25:4
Flavonoid glycoside n-BuOH–HOAc–H2O, 65:15:25
n-BuOH–HOAc–H2O, 3:1:1
 EtOAc–MeOH–H2O, 50:3:10
EtOAc–MeOH–HCOOH–H2O, 50:2:3:6
EtOAc–EtOH–HCOOH–H2O, 100:11:11:26
EtOAc–HCOOH–H2O, 9:1:1
EtOAc–HCOOH–H2O, 6:1:1
EtOAc–HCOOH–H2O, 50:4:10
EtOAc–HCOOH–HOAc–H2O, 100:11:11:26
EtOAc–HCOOH–HOAc–H2O, 25:2:2:4
THF–toluene–HCOOH–H2O, 16:8:2:1
Chloroform –MeCOMe–HCOOH, 50:33:17
Chloroform –EtOAc–MeCOMe, 5:1:4
Chloroform –MeOH–H2O, 65:45:12
Chloroform –MeOH–H2O, 40:10:1
MeCOMe–butanone–HCOOH, 10:7:1
MeOH–butanone–H2O, 8:1:1
Flavonoid glucuronide EtOAc–Et2O–dioxane–HCOOH–H2O, 30:50:15:3:2
EtOAc–EtCOMe–HCOOH–H2O, 60:35:3:2
Flavanone aglycone CH2Cl2–HOAc–H2O, 2:1:1
Flavanone glycoside Chloroform –HOAc, 100:4
Chloroform –MeOH–HOAc, 90:5:5
n-BuOH–HOAc–H2O, 4:1:5 (upper layer)
Chalcones EtOAc–hexane, 1:1
Isoflavones CHCl3–MeOH, 92:8
Chloroform –MeOH, 3:1
Isoflavone glycoside n-BuOH–HOAc–H2O, 4:1:5 (upper layer)
Dihydroflavonol Chloroform –MeOH–HOAc, 7:1:1
Biflavonoid Chloroform –MeCOMe–HCOOH, 75:16.5:8.5
Toluene–HCOOEt–HCOOH, 5:4:1
Anthocyanidin dan EtOAc–HCOOH–2 M HCl, 85:6:9
anthocyanin n-BuOH–HOAc–H2O, 4:1:2
EtCOMe–HCOOEt–HCOOH–H2O, 4:3:1:2
EtOAc–butanone–HCOOH–H2O, 6:3:1:1
Proanthocyanidin EtOAc–MeOH–H2O, 79:11:10
EtOAc–HCOOH–HOAc–H2O, 30:1.2:0.8:8

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3 komentar:
1.

yulia15 Oktober 2012 06.33

results of methylation or acetylation of flavones and flavonols require
nonpolar solvents such as chloroform-methanol (15:1). Being such
flavonoid aglycone apigenin, luteolin and quercetin can be separated
 with chloroform methanol (96:4) or with the same polarity.

why the results of methylation or acetylation of flavones and flavonols

require nonpolar solvents in separation? then why comparisons
chloroform-methanol used in the separation of flavonoid aglycone
greater than the flavones and flavonols methylation results?
Balas

2.

Aminah Alhabsyi15 Juni 2017 20.08

Reference please
Balas

3.

boc Sciences16 Desember 2018 06.28

Complanatuside A, a flavonoid compound, is isolated from Astragalus

complanatus and Lysimachia christinae. Astragalus complanatus
R.Brown is a widely used herbal material in traditional Chinese
medicine. The total flavonoid (TF) is an active fraction extracted from
seeds of Astragalus complanatus R.Brown. Complanatuside A
Balas

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