Pharmacognostic and Preliminary Phytochemical Screening of Cichorium intybus Linn Seed

 

Tauseef Shaikh, Atar Mujum , Khan Wasimuzzama and Rukhsana A Rub*

Department of Pharmacognosy, M.C.E. Society^’s Allana College of Pharmacy, Pune-411001, MS, India

*Corresponding Author E-mail: hailrukhsar@yahoo.com

ABSTRACT:

Cichorium intybus a member of Asteraceae family, is a perennial herb cultivated throughout India. It is commonly known as Chicory. It has been reported to have antifungal, antitibacterial, hypoglycemic and hepatoprotective activity in various traditional systems of medicine. It is also found to have antineoplstic, antioxidant, analgesic and sedative activity. As the Pharmacognostic and Phytochemical data on the seeds of Cichorium intybus have not been reported previously, various pharmacognostic parameters like, examination of macroscopical, microscopical characters, ash values, extractive values, foreign organic matter and LOD were studied with the aim of establishing monograph for this species. Phytochemical screening included qualitative chemical examination and Thin layer chromatography of the detected phytoconstituents. Microscopical characters showed the presence of parenchyma, single layered tangentially elongated epidermis, well defined endosperm and papus. Powder characteristic also showed the presence of epidermal cells, papus and endosperm. The total ash, water soluble ash and acid insoluble ash were found to be 9.2%w/w, 1.1%w/w, 1.4%w/w respectively. The moisture content was found to be 7 w/w. Water, Alcohol and Chloroform soluble extractive values were found to be 12.2%w/w,13.4%w/w, 1.2%w/w respectively. Preliminary phytochemical investigation of aqueous and methanolic extract of seeds of cichorium intybus revealed the presence of Carbohydrates, Alkaloids, Tannins and Flovonoids which were further confirmed by thin layer chromatography with the Rf values as 0.94,0.75 and 0.78 for carbohydrates, alkaloids, flavonoids and 0.81 and 0.90 for tannins and flavonoids respectively.

 

 

 

1. INTRODUCTION:

Chicory^1-3 a member of asteraceae family, commonly known as Chicory, is a perennial erect herb, usually 30-90 cm in height with a fleshy tap root up to 75cm in length. Roots are brownish yellow outside and white inside. The fruit is small, about 3 mm long, and 2mm broad with a crown of about 0.5m long, having a ring of white or straw colour. Leaves are broadly oblong, oblanceolate or lanceolate, crowded at the base forming rosette arranged spirally on the stem. Seeds are about 2.5mm long, ovoid, apex pointed, tip brownish^13,15. It is one of important medicinal plant cultivated throughout India. The plant has been known to possess hy-poglycemic⁷, antineoplstic⁸, antioxidant⁹, analgesic, sedative¹⁰, antifungal¹¹, antitibacterial¹² and hepatoprotective¹³ activity.

 

The literature survey suggests that inspite of numerous medicinal uses attributed to this plant, it is not much explored pharmacognostically and no such type of extensive work with respect to its phytochemical account is seen. Therefore, the present investigation is an attempt to explore the plant for its pharmacognostic, phytochemical and chromatographic profile.

 

2. MATERIALS AND METHODS:

The seeds of Cichorium intybus were collected from Ayurvedic shop of Pune, India. The botanical identity of the plant was confirmed by Botanical Survey of India, Pune (BSI/WRC/Tech/2010/1008).

 

2.1. Macroscopic and Microscopic examinations: ^5,6,14

Macroscopical features like colour, odour, taste, size, shape, apex were carefully observed. Microsc- opical studies were carried out by preparing a thin transeverse section of the seed. The section was cleared, stained with Phloroglucinol and Hydrochloric acid and mounted in glycerin and observed under microscope using 10X lense. For powder characteristics, the same staining reagent was used.

 

2.2. Proximate analysis: ⁴

The Physicochemical analysis such as Ash values (Total ash, Acid insoluble and Water soluble Ash values), Foreign matter, Moisture  content, Extractive values (Water, Alcohol, Chloroform) were carried out as per Pharmacopoeial guidelines^.

 

2.3. Preliminary Phytochemical Screening: ²

The dry seeds were pulverized and extracted by continuous hot extraction (Soxhlet) method using Water, Methanol, Chloroform and n-hexane. The extracts were concentrated and subjected to phytochemical screening using standard procedures. The compounds were analysed for Alkaloids, Glycosides, Flavonoids, Tannins, Carbohydrates, Proteins, Amino acids and Steroids.

 

2.4. Thin Layer Chromatography:³

Thin Layer Chromatography of different extracts of seeds of Cichorium intybus were carried out. The resolution of TLC plates was tried in different solvent systems and best resolving solvent systems as mentioned in Table 3 and Table 4 were used for running the plates. The plates were then exposed to various detecting reagents (Table 3 and 4) or observed under U.V. light.

 

3. RESULTS AND DISCUSSION:

3.1. a. Macroscopical characteristics:

The morphological features like light brown colour, characteristic odour, bitter taste, ovoid shape with the length of 2.1mm and pointed apex were comparable with the standard information.

 

3.1.b. Microscopical characteristics:

Microscopical studies showed the presence of parenchyma, single layered epidermis, endosperm and papus. Fig (1).

 

Fig. 1: Transeverse Section of Cichorium intybus seed

 

3.1.c. Powder charecteristics:

The grayish yellow coloured powder showed the presence of tangentially elongated thick walled epidermal cells, a well defined endosperm and papus Fig. (2).

 

Endosperm                                          Epidermal cells

 

Papus

 

Fig. 2: Powder microscopy of Cichorium intybus seed

 

3.2. Proximate analysis:

Table 1: Physicochemical constant for Cichorium intybus seeds

Physicochemical parameters	Values % (w/w)
Total ash	9.2
Water soluble ash	1.1
Acid insoluble ash	1.4
Alcohol soluble extractive value	13.4
Water soluble extractive value	12.2
Chloroform soluble extractive value	1.2
Loss on drying	7
Foreign matter	4

 

3.3. Preliminary phytochemical screening:

Preliminary phytochemical screening revealed the presence of Carbohydrate, Alkaloid, Tannins and Flavonoids (Table 2).

                   

 

Table.2: Preliminary Phytochemical screening of Cichorium intybus seeds

Sr. No	Tests	Water extract	Chloform exrtract	Methnol extract	n-hexane extract
1	Carbohydrates a)    Molisch Test b)    Fehilings Test c)     Benedicts Test d)    Barfoed’s Test	+ + + _	_ _ _ _	_ _ _ _	_ _ _ _
2	Proteins a)    Biuret Test b)    Millions Test	_ _	_ _	_ _	_ _
3	Amino acids a)    Ninhydrin Test	_	_	_	_
4	Steroids a)    Salkowski Test b)    Liebermann – Burchard reaction	_ _	_ _	_ _	_ _
5	Glycoside a)    Killer-Killani Test b)    Legal’s Test c)     Baljet test d)    Borntrager’s Test e)     Modified Borntrager’s Test	_ _ _ _ _	_ _ _ _ _	_ _ _ _ _	_ _ _ _ _
6	Alkaloids a)    Dragendroff’s Test b)    Wagner Test c)     Mayers Test d)    Hagers Test	+ + + +	_ _ _ _	_ _ _ _	_ _ _ _
7	Flavonoids Shinoda test	+	_	+	_
8	Tannins a)    5% Ferric Chloride Test b)    Lead Acetate Test	_ _	_ _	+ +	_ _

+  indicates presence of constituents,    -  indicates absence of constituents.

 

3.4. Thin layer chromatography:

Table.3:Thin layer chromatography profile for Aqeous extract of Cichorium intybus seeds:

Compounds	Carbohydrate	Alkaloid	Flavonoids
Stationary phase	Silica Gel	Silica Gel	Silica Gel
Mobile phase	Benzene:glacial aceticacid:methanol (20:20:60)	Chloroform:methanol: Diethylamine  (80:20:10)	Ethyl acetate:formic acid:glacial acetic acid (100:11:11:26)
Visualising agents	Anisaldehyde sulphuric acid	10%Ethanolic sulphuric acid	Anisaldehyde sulphuric acid
Rf values	0.94	0.75	0.78

 

Table.4:Thin layer chromatography profile for Methanolic extract of Cichorium intybus             seeds:

Compounds	Tannins	Flavonoids
Stationary phase	Silica Gel	Silica Gel
Mobile phase	Ethyl acetate: glacial acetic acid:water (90:10:10)	Ethyl acetate:formic acid:glacial acetic acid (100:11:11:26)
Visualising agents	5%FeCl3 solution	Anisaldehyde sulphuric acid
Rf values	0.81	0.90

 

 

DISCUSSION:

Microscopical examination of the seeds of Cichorium intybus showed a unique and well defined cellular structure. Transverse section showed presence of parenchyma, single layered epidermis, endosperm and pericarp Fig (1).

Microscopy of powder showed presence of epidermal cells, endosperm and papus. Fig. (2).

 

The proximate analysis is useful for setting standards for crude drug and for detecting adulteration. The ash values are important for determination of purity of drug. The extractive values are primarily useful for determination of exuasted or adulterated drug. Proximate analysis showed satisfactory results with respect to foreign matter, moisture content and ash values etc. The water soluble extractive value was found to be 12.2% whereas alcohol soluble extractive value and chloroform soluble extractive value were found as 13.4and 1.2% respectively (Table 1).

i)TLC for alkaloid      ii)TLC for Carbohydrates     iii)TLC for Flavonoids

 

Fig.3.TLC for Water extract:

 

In the phytochemical investigations of the seeds of Cichorium intybus, the aqueous extract showed presence of Alkaloids, Flavonoids, Carobohydrates, which were further confirmed by Thin Layer Chromatography with the Rf values for Alkaloid (0.75), Flavonoids (0.78) and Carbohydrate (0.94) (Fig. 3).

                   

i)TLC of Tannins          ii)TLC of Flavonoids

 

Fig.4.TLC for Methanolic extract:

Methanolic extract of the seeds of Cichorium intybus showed presence of Tanins and Flavonoids which were also further confirmed by TLC with Rf values for Tanins and Flavonoids as 0.81 and0.90 respectively (Fig.4).

 

The quantitative estimation of some of the pharmaconostic parameters is useful for setting standards for crude drugs. As no such work, which can provide better understanding regarding the drug was done earlier; present research work can prove to be an important data for the standardization of the drug and could be helpful in establishing a monograph of the plant.

 

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Received on 19.05.2010        Modified on 12.06.2010

Accepted on 24.06.2010        © AJRC All right reserved