SEPARATION AND IDENTIFICATION OF FLAVONOIDS FROM MATRICARIA CHAMOMILLA BY TLC. [615907]

SEPARATION AND IDENTIFICATION OF FLAVONOIDS FROM MATRICARIA CHAMOMILLA BY TLC.

Abstract : Matricaria chamomilla is one of the most popular herbs is the grasses, flavored commoun
in all areas of the plains, meadow at the edge of the road in summy spots or on land . The main active
flavonoids are: rutina,miricetina, quercetin and hyperoside . These flavonoids act as powerful
antioxidants .
Matricaria chamomilla has a leading role in promoting cardiovascular health. This includes an
improvement of coronary ar terial blood flow and supply of oxygen to the heart, control . He has
studied the content of flavonoids in leaf extract of Matricaria chamomilla .
Key words: Matricaria chamomilla the leaves and flowers , active flavonoids ,TLC.

INTRODUCTION:

Currently, chromatographic methods have a universal diffusion , both theoretical and practical and
have the highest efficacy of all methods separat e used in analytical chemistry and technology .[2]
Chromatographic methods can be classified according to the following crite ria: [3,7] – the nature of
mobile and stationary phase
– nature elementary sorption process
– sorption isotherm of form basic process
Matricaria chamomilla grows as a thorny shrub with white and pink flowers and berries in Europe,
Western Asia, North Am erica and North Africa . Parties used in the pharmaceutical industry are leaves
and flowers , the fruits are used especially in traditional preparations. [5,6].
The main active flavonoids are: rutina, miricetina, quercitin and hyperoside . These flavonoids act as
powerful antioxidants . Hawthorn has a primary role in promoting cardiovascular health . This includes
an improvement in coronary artery blood flow and oxygen supply to the heart such contractions of
the heart muscle , stable angina , congestive heart fai lure.
EXPERIMENTAL DATE

We studied the flavonoid content of leaf extract of Matricaria chamomilla . Was done by thin layer
chromatography (TLC ) separation and identification their off. [1, 8].
The experimental procedure were: e xtraction (1) and s eparatio n and identification of the main
flavonoids (2).

1) Extraction of flavonoids
1,5 g the dried herb was extracted with 1 5 ml of 1% HCl. Both of flavonoids and extract contains
proteins. To remove the protein is added a solution of 10 % sodium phosphotungstic Na3 P (W 3O10
)4 when it precipitates . After centrifugation , the solution containing flavonoids passing over a
column of ion exchange resin (Amberlite R 120 ). Elute column with 50 ml of 10% ammonia . The
solution obtained is evaporated to dryness on a wate r bath . For the subsequent operations of flash
residue is taken up with 1 ml 30% isopropanol .
We developed a new method for the separation and identification of flavonoids by TLC . The results
were tabulated in Table 1.
2) Separation and identification of flavonoids
Separation and identification of flavonoids was done by two-dimensional thin-layer chromatography
plates were used with microcrystalline cellulose .
Preparation of cellulose sheets is as follows : weigh 15 g of powdered pharmaceutical balance of
microcrystalline cellulose (Merck ), to which was added 30 ml of water and 5 ml of methyl alcohol .
If the cellulose suspension is prepared in water alone, after drying, the adhesive layer thereof on an
inert support glass crack, which prevents the migration o f the mobile phase system formed by

interrupting the capillary during the stationary phase. The formed suspension was subjected to
homogenization using a magnetic stirrer for 10 minutes at warm.
Resulting homogeneous paste is spread on the machine stretche d thin, the plate 20 * 20 cm, which
were previously degreased for snorting stationary phase on inert support. The plates thus obtained
were allowed to dry at room temperature, for a perfectly flat surface, for 24 hours.
The thickness of the stationary phas e (microcrystalline cellulose) was adjusted to 0.3 mm. At a
distance of 2 cm from one end of the plate can thus be applied with a micro syringe samples
analyzed plant extracts. For hawthorn was used as the mobile phase solvent mixture butanol:
acetone: ace tic acid: water = 35: 35: 7: 23 (by volume). This mixture was homogenized well by
shaking, and is left at room temperature for 24 hours to separate the phases. Mixture directly into
the separating funnel, and after 24 hours separates two distinct phases .
The organic phase used for the elution is evident saturated with water. Attempts to optimize the
separation of flavonoids by two -dimensional TLC. For elution in the first direction to use the system
butanol: acetone: acetic acid: water = 35: 35: 7: 23. Afte r drying the plates was done elution in the
second direction using a solvent mixture: methanol: water: pyridine = 80: 20: 5. Elution was done in
the direction perpendicular to the first. Both elution was done two times.

Fig.1 . TLC chromatogram separation of flavonoids : quercetin , hyperoside , rutina, miricetina

For viewing using a ninhydrin solution obtained by dissolving 3.6 g of ninhydrin in 50 ml of a mixture
of butanol: acetone (50: 50 v / v). Ninhydrin solution was sprayed using a spray particular,
homogeneous on the whole surface of the plate. In both cases, after elution, the plates were left at
room temperature for several hours after that enter the 110 C. 15 minutes of stand on white
background pink appear distinct areas – reddish of flavonoids p resent in the test sample.

CONCLUSIONS

All the main flavonoids present in the plant, using thin layer chromatography, plant with multiple
uses in pharmacology were identified and separated .
The identify was based on retention times
The chromatogram show s that each of flavonoid present in the extract, corresponding to this peak
has increased in proportion to the contribution of the concentration of each extract
Some components appear strongly colored, others less strong component related to sensitivity
reaction given reagent used