Pharmacognostic and Preliminary Phytochemical Screening of Sesbania grandiflora root

 

Keerthi Prashanth H V1, R Nandeesh1, Kaveri H V2, Vidyashree M2, Manjunatha E1,

Veeresh P Veerapur1, Pramod P Desai2, S Vijayakumar1

1Sree Siddaganga College of Pharmacy, Tumkur, Karnataka, India.

2Department of Botany, Tumkur University, Tumkur, Karnataka, India.

*Corresponding Author E-mail: rnandeesh2005@gmail.com

 

ABSTRACT:

Sesbania grandiflora belongs to Family: Fabaceae is extensively distributed in Karnataka, West Bengal, Assam and North-Eastern parts of India. Sesbania grandiflora has the common names of Agati, Corkwood Tree and West Indian Pea. Sesbania grandiflora is used as Anticancer, Antioxidant, Cardioprotective, Antiulcer, Hepatoprotective Anti-inflammatory etc. The present study intended with various phytochemical screening and pharmacognostic studies on Sesbania grandiflora. Extraction withsoxhelt extraction apparatus and macro-microscopical parameters are carried out. The anatomical characteristics were observed and photographed by taking various sections of root. Preliminary phytochemical evaluation of the aqueous extracts revealed that presence of carbohydrate, proteins, flavonoids, alkaloids, tannins and glycosides. Pharmacognostic investigations like Ash value, Fluorescence analysis, Thin layer chromatography of the root extract performed in the present study to exhibit quick identification of the drug and are particularly useful in the case of powdered forms and also contribute towards establishing Pharmacopoeial standards for the specified plant.

 

KEYWORDS: Sesbania grandiflora, Anticancer, Antioxidant, Cardioprotective, Antiulcer, Hepatoprotective, Ash value.

 

 


INTRODUCTION:

Herbal drugs being used as medicines since ancient period. The increasing use of herbal drugs and concerns over their safety and efficacy has certainly augmented the need of standardization of these herbal drugs. World Health organisation (WHO) has set up guidelines for standardization of these drugs, which are used as a standard by the majority of countries1

 

The other scientific names of Sesbania are Robinia grandiflora Linn, Aeshynomene grandiflora Linn, Sesban grandiflora Poir, Agatig randiflora (L.) Desv. Locally it is known as caturay, katurai (Chamorro), corkwood tree, scarlet wisteria, sesban, vegetable hummingbird (English), agati a grandesfleurs (French), ohaike‘oke‘o (Hawaiian), katurai (Palauan), sepania (Samoan), afai, ofai,ouai, oufai (Tahitian), agathi, agati (Tamil), hadga (Hindi, Marathi)2

 

Sesbania grandiflora is belongs to family Fabaceae a small erect quick-growing short-lived soft-wooded treesparsely branched. Bole straight and cylindrical, with the wood is white in colour andsoft. The tree grows generally 5 to 12 meters in height along with leaves 20 to 30 centimetres long, and pinnate having 20 to 40 pairs of leaflets, whichare 2.5 to 3.5 centimetres long. The flowers are white and 7 to 9 centimetres long. The pods are linear, 20 to 60 centimetres long, 7 to 8 millimetres wide, pendulous, and somewhat curved, and contain many seeds3

 

This plant is used in Siddha system of Indian traditional medicine for the treatment of a wide spectrum of ailments including anaemia, bronchitis, fever, headache, ophthalmic, nasal catarrh, inflammation, leprosy, gout and rheumatism. It also possesses anxiolytic and anticonvulsive and hepatoprotective properties. In addition, S. grandiflora is mentioned as a potent antidote for tobacco and smoking-related diseases. However, the mechanisms underlying its beneficial effects against chronic smoking associated diseases are yet to be determined. The various parts of Sesbania are used as medicine for many diseases and disorders4

 

BOTANICAL CLASSIFICATION:

Common name   :Vegetable humming bird, agati or humming bird tree.

Botanical name   :  Sesbania grandiflora

Family                   :  Fabaceae

Species                  :  S. grandiflora

Hindi                      : Agast, Basna, Hadga, Hathya

Kannada               : Agasi, Agaci.

English                   : Vegetable hummingbird

Telugu                   : Avesi, Avaasinara

Bengali                  : Agathi, Agate, Agusta, Bagphol

French                   : Poisvalliere, Crest de gallo, Pico

 

MATERIALS AND METHODS:

Collection of plant material and storage:

Sesbania grandiflora root was collected during the month of February first week from the local regions like Obalapura village, Pavagadataluk, Tumakuru District, Karnataka. The Specimen was identified and authenticated by Dr. Y N Seetharam, Chief Scientific Officer, Vriksha Vijnan Pvt. Ltd, Bengaluru.

 

Preparation of Plant Extraction:

The roots of Sesbania grandiflora were chopped into smaller pieces and dried under shade at normal temperature for seven days, then it was powdered in a mixer grinder and further used in soxhelt extraction process5 58 g of powder sample was weighed along with 300ml of different solvents (Petroleum ether, Chloroform, Methanol and Water) were loaded into the soxhelt apparatus at their corresponding boiling points and run continuously for 6-8 hours respectively.

 

After 6 hours the desired compounds was concentrated in the distillation flask. After extraction the solvents was removed, typically by means of rotary evaporator, yielding the extracted compound. The non soluble portion of the extracted solid which remained in the thimble was discarded. Finally, the extract was collectedin different colours and yield was noted.

 

Pharmacognostic studies:

The plant was studied for morphological characters including size, shape, colour, odour, taste, and other features. Microscopical study was performed for both entire (free hand T.S, T.L.S, and R.L.S sections) and powdered material using chloral hydrate as clearing agent, phloroglucinol, concentrated HCl and glycerine as staining agents6. The anatomical characteristics were observed and photographed analytically using stereo zoom digital microscope (Lynx Company, Model No-1601052).

 

Powder analysis:

Root powder of the Sesbania grandiflora was observed under compound microscope which was stained with phloroglucinol, chlorohydrate, concentrated HCl and glycerine, pictures as per the protocol followed by Chase et al., 1948.

 

Fluorescence analysis:

The fluorescence character of the plant powder was studied under UV light (366 nm) and after treatment with different reagents like Hydrochloric acid, Sodium hydroxide, Nitric acid, Sulphuric acid, Ferric chloride, Methanol, ethyl acetate, petroleum ether, ammonia, chloroform and potassium hydroxide5

                                                                                                                                                                                

Preliminary Phytochemical Studies:                           

The following experiments were conducted on the root extracts of Sesbania grandiflora and their respective chemical constituents fractions were tested for the presence of different secondary metabolites. The different extracts obtained in our study after each successive solvent extraction was qualitatively tested for the presence of various phytochemicals. Like Alkaloids, Glycosides, Tannins, Carbohydrates, Steroids, Saponins, Flanoids and Phenols. The preliminary phytochemical screening was carried out6

 

Determination of Ash values:

The ash remaining after complete ignition of the plant materials is determined by three different methods known as Total ash, Acid-insoluble ash and Water soluble ash using suitable methods5

 

Thin layer chromatography (TLC):

In this study, thin layer chromatography (TLC) technique was used to separate mixtures. TLC was performed by using a uniform layer of silica gel which acted as a stationary phase (solid matrix) and mobile phase was used as a combination of different organic solvents7

RESULTS AND DISCUSSION:

Extraction:

The successive extraction was carried out using various solvents like Petroleum ether, chloroform, methanol and water. The physical property and the percentage yield were shown in table no.1

 

Pharmacognostic studies:

Here in this study, the microscopic characters when observed which revealed the presence of typical root characteristics as Periderm which consists of cork, phellogen and phelloderm. In our sample, cork was observed dark brown in colour. It is composed of multiple layers, with narrow and tangentially elaongated cells. Phellogen and Phelloderm were observed one beneath the other and are compactly arranged layers of cells. These are 2-3 layered cells which are blue in colour. The periderm then originates in cortex region. The cortex region is found just below the periderm layer. These are thin walled parenchynatous cells. It showed absence of starch grains in the parenchymatous cells (Figure 1). Phloem is observed just above the cambium. The Secondary xylem showed presence of lignified xylem vessels with absence of pith. The medullary rays run radially from the xylem up to one to five cells in width. Xylem parenchyma is moderately thick walled, lignified with pitted walls.

 

Powder analysis:

In this methodology, root powder of the Sesbania grandiflora were observed under microscope stained with phloroglucinol, chlorohydrate, concentrated HCl and glycerine. Powder of root is off-white, non aromatic, bitter. The microscopic examination of the powder showed fragments of parenchymatous cells, elongated cork cells, cortical cells, xylem vessels and pitted vessels with simple perforation was observed (Figure 2).

 

Fluorescence analysis:

 In this technique, the fluorescence character of the plant powder was studied under UV light (366 nm) which showed the different colour with various test solutions as shown in table no 2.

 

Phytochemical screening:

Phytochemical screening of the plant extracts of S. grandiflora was made by running the root through solvents such as Petroleum ether, Chloroform, Methanol and water. The extract of root show positive tests for alkaloids, steroids, carbohydrates, glycosides, amino acids and proteins, saponins, flavones, anthroquinone glycosides and phenolic compounds. (Table 3)

 

Determination of Ash values:

The ash values of the Sesbania grandiflora root extract were determined with total ash content value, acid insoluble ash value and water soluble ash value as represented in table 4.

 

Thin layer chromatography:

The preliminary phytochemical screening of the different solvent extracts of (petroleum ether, chloroform, methanol and water) revealed the presence of chemical constituents like Alkaloids, Tannins and phenolic compounds, Flavonoids, Carbohydrates and Amino acids which was confirmed by performing TLC separation technique using UV- radiation at 365 nm. Presence of alkaloids and flavonoids was detected visually by UV- radiation at 365 nm. Alkaloids were confirmed by Lightest green and blue fluorescence where as flavonoids were confirmed by yellow fluorescence. (Table 5 and figure 3)

 

Most of the traditional knowledge about medicinal plants in the form of oral knowledge. There fore it is necessary to maintain documentation and systematic regulations for their wide spread application in healthcare system. Accordingly, in this study few herbal standardisation techniques like microscopic evaluation, powder analysis, fluorescence analysis and preliminary phytochemical analysis and chromatographic techniques were carried out for this plant root to develop qualitative parameters as per herbal pharmacopoeial standards to authenticate S. grandiflora.

 

CONCLUSION:

India has a rich medicinal flora and largest producer of medicinal herbs, but the major hurdle is lack of standardisation investigation to meet the global herbal market. Hence this study is quite possible to set the standards of selected plants as per the pharmacopoeial guidelines and it is useful for the selection of proper medicinal plant.


 

 

Table 1: Quantitative determination of extractive value of root extractsof S. grandiflora

Sl. No

Extracts

State

Colour

Odour

% of yield

1.

Petroleum ether

Sticky solid

Light brown

Characteristic

0.461%

2.

Chloroform

Sticky solid

Chocolate brown

Characteristic

0.512%

3.

Methanol

Semi solid

Dark Brown

Characteristic

6.74%

4.

Water

crystalline

Black

Characteristic

2.06%

N. B:- Formula Used: - Percentage of extractive yield (W/W) = Weight of dried extract/ Weight of dried root × 100

 

 

 


Table 2. Shows the fluorescence analysis of Sesbania grandiflora root:-

SI. No

Tests

Color observed under the day light

Color observed under UV-(365 nm) radiation

1.

Powder + (alc) 1N NaOH

Dark brown

Creemish

2.

Powder +(aq) 1 N NaOH

Brown

Greenish

3.

Powder + picric acid

Yellow

Black

4.

Powder + Ammonia

Light yellowish to brown

Green

5.

Powder +ethylacetate

_

Lightish green

6.

Powder +dil. 50% H2SO4

_

Lightish green

7.

Powder + pet ether

_

_

8.

Powder+50% HCL

Brown

Black

9

Powder + methanol

_

Lightish green

10.

Powder + chloroform

_

Green

11.

Powder + con. Nitric acid

Reddish brown

Black

12.

Powder+ KOH

Brown

Dark green

 

Table 3. Phytochemical studies of Sesbania grandiflora root extracts

Test

Pet. Ether extract

Chloroform extract

Methanolic extract

Water extract

1.      Alkaloids

 

a)      Dragendroff’s Test

+

+

+

+

b)      Mayer’s Test

_

+

+

+

c)      Wagner’s Test

+

+

+

+

d)      Hager’s Test

+

+

+

+

e)      Tannic acid Test

_

_

+

_

f)       Murexide Test

+

+

+

_

2.      Carbohydrates

 

a)      Mollisch’s Test

+

+

+

_

b)      Fehling’s Test

+

+

+

+

c)      Benedict’s Test

+

+

+

+

3.      Hexose sugar

 

a)      Selwinoff’s Test

+

+

+

+

4.      Non-reducing polysaccharides

 

a)      Iodine Test

_

_

_

_

Mucilage Test

­

+

+

+

Amino acids and protein Test

 

a)      Biuret Test

_

_

_

_

b)      Million’s Test

_

_

_

_

c)      Xanthoprotin Test

_

_

_

_

d)      Ninhydrine Test

_

_

_

_

5.      Steroids

 

 

 

 

a)      Salkowski’s Test

+

_

_

_

b)      Liebermann Burchard’s Test

_

_

_

_

6.      Glycosides

 

a)      Baljet’s Test

+

_

_

_

b)      Legal’s Test

+

+

+

+

7.      Antheranquinone glycoside

 

Borntrager’s Test

_

_

_

_

a)      Foam Test

_

_

_

_

a)      Coumarins Test

_

_

_

_

8.      Flavonoids

 

 

 

 

a)      Shinoda Test

+

+

+

+

b)      Sulphuric acid test

+

+

+

+

9.      Tannins and phenolic compounds

 

 

 

 

a)      Fecl3 Test

_

_

+

+

b)      Lead acetate Test

+

+

+

+

c)      Bromine water Tests

+

+

+

+

++ = (strong detected) + = (weakly detected) - = (no detection)

 


Table 4: Physicochemical analysis of Sesbania grandiflora root

Ash values

Percentage of yield

Total ash

2.48%

Acid insoluble ash

0.0%

Water soluble ash

0.015%

 

 

Table 5: TLC analysis of Secondary Metabolites Sesbania grandiflora root

Sl. No

TLC for alkaloids

Different solvent systems

Extracts of different solvents

No. of spots

Rf values

1.

 

chloroform: methanol: acetic acid (18:1:1)

Petroleum ether

3

0.34, 0.86, 0.98

2.

 

 

Chloroform

7

0.17, 0.30, 0.44, 0.55, 0.74, 0.89, 0.96

3.

 

 

Methanol

6

0.13, 0.20, 0.29,0.46,0.65, 0.86

4.

 

 

Water

-

-

5.

 

toluene: ethylacetate: diethylamide (17.5:5:2.5)

Petrollium ether

5

0.25, 0.45, 0.54, 0.64, 0.79

6.

 

 

Chloroform

6

0.16, 0.25, 0.33, 0.48, 0.61, 0.72

7.

 

 

Methanol

4

0.19, 0.30, 0.53, 0.74

8.

 

 

Water

-

-

9.

TLC for Flavonoids

ethylacetate: formic acid: glacial acetic acid: water (10:1.1:1.1:2.6)

Petrollium ether

2

0.33, 0.93

10.

 

 

Chloroform

4

0.33, 0.55, 0.81, 0.95

11.

 

 

Methanol

4

0.33, 0.55, 0.76, 0.96ss

12.

 

 

Water

-

-

 


 

Figure 1: a) T.S of S. grandiflora root b) cortex region c) absence of pith d) T.L.S of root e) R.L.S of root f) absence of starch.

 

 

Figure 2:- Root powder of the Sesbania grandiflora were observed under microscope a)Elongated cork cells b) Conducting tissues surrounded by parenchymatous cellsc) Cork cells d) Xylem vessels e) Pitted vessels with simple perforation f)cortical Cells

 


 

Figure 3: TLC for alkaloids SS1 (a), SS2 (b) and for flavonoids SS3 (c) Different solvent extracts are used (ABC) 1 (Pet. ether), 2 (chloroform), 3 (methanol), 4(water)


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Received on 08.08.2018          Modified on 26.08.2018

Accepted on 19.09.2018  ©A&V Publications All right reserved

Res. J. Pharmacognosy and Phytochem. 2018; 10(4): 285-290.

DOI: 10.5958/0975-4385.2018.00046.8