Universities Research Journal 2011, Vol. 4. No.

Chemical Investigation and Antimicrobial Activities of

Sesbania grandiflora L.
Aye Aye Aung

Abstract
Sesbania grandiflora L. belongs to the family, Fabaceae, order Fabales. It
is commonly known in Myanmar as Pauk-pan-phyu and Pauk-pan-ni. The
plants were collected from Hpa-an Township, Kayin State and prepared
for chemical studies in this work. According to the phytochemical
investigation, alkaloid, α-amino acid, carbohydrate, starch, reducing sugar,
glycoside, phenolic compound, saponin, tannin, terpenoid/steroid and
flavonoid were found to be present and cyanogenic glycoside were absent
in these plants. In physicochemical studies, the methanol and ethanol
soluble matter content were found to be greater than soluble matter
content of other organic solvents in leaves. These plants analysis are to
determine the abundance amount of elements in Sesbania grandiflora L.
(Pauk-pan-phyu and Pauk-pan-ni) which are a quantitative indication of
the level of nutrients in plant and is obtained by using EDXRF (Energy
Dispersive X-ray Fluorescence) spectrometer. In this experiments,
chloroform extracts of leaves showed more significant activity against all
test organisms especially on Pseudomonas aeruginosa.
Key words: Sesbania grandiflora L., Qualitative and Quantitative analysis,
Antimicrobial Activities.

Introduction
Sesbania grandiflora L. belonging to Fabaceae commonly known as
Sesbania which is often planted for its edible flowers and pods in tropical
countries. It was known as Pauk-pan-phyu and Pauk-pan-ni in Myanmar. It
is believed to have originated either in India or Southeast Asia and grown
primarily in hot and humid areas of the world. From present generation, it is
a green manure crop useful in paddy cultivation; it is one of the most
valuable medicinal herbs. The tree is grown as an ornamental shade tree,
and for reforestation. (Watt, 1972 and James, 1983). The crude methanolic
extracts of the flowers, seeds, rachis and leaves of Sesbania grandiflora
were strongly positive for saponin (by Fehling’s A and B test). The flowers,
seeds and leaves were slightly positive for alkaloids. Sterols were markedly
found in the rachis and leaves; strongly present in the flowers; and slightly
present in the seeds. The rachis and leaves contained plenty of tannins

Demonstrator, Department of Botany, East Yangon University

338 Universities Research Journal 2011, Vol. 4. No. 1

(Rastogi, 1969).This paper deals with chemical investigations including

phytochemical tests, physicochemical characterization, elemental analysis,
nutritional values.

Materials and Methods

Collection, classification and identification of Sesbania grandiflora L.
Sesbania grandiflora L. plant parts were collected from Hpa-an
Township, Kayin State, during the months of July to December in 2009.
The dried samples were made into fine powder using a mortar and pestle
and stored in air tight container to prevent moisture and other contamination.
Qualitative and quantitative analysis of Sesbania grandiflora L.
Qualitative analysis
Qualitative analysis concerned with the presence of alkaloids, amino
acid, carbohydrate, starch, reducing sugar, cyanogenic glycoside, glycoside,
phenolic compound, saponin, tannin, steroid and flavonoid were
investigated by the methods of British Pharmacopoeia (1968), Trease and
Evans (2002). Preliminary phytochemical examination was carried out in
the phytochemical laboratory of Chemistry Department, University of
Yangon.
Quantitative analysis
Quatitative analysis including moisture content, total ash, acid
soluble, water soluble ash, acetone soluble, chloroform soluble, petroleum
ether soluble, ethyl acetate soluble, ethanol soluble, methanol soluble and
water soluble contents of leaves were determined by the methods of in B P,
(1968).
Elemental analysis and nutritional values of Sesbania grandiflora L.
Elemental analysis of leaves
The relative concentration of elements in Sesbania grandiflora L.
powdered leaves were analyzed by using Energy Dispersive X-ray
Fluorescence (EDXRF) spectrometer, Shimadzu Co. Ltd., Japan, at the
Universities’ Research Center, University of Yangon.
Some nutrient elements such as Copper (Cu), Potassium (K),
Calcium (Ca), Iron (Fe) and Phosphorus (P) in the leaves were also
quantitatively analyzed by A A Analyst.800 Atomic Absorption
Universities Research Journal 2011, Vol. 4. No. 1 339

Spectrophotometer England (A A S) at the Universities’ Research Center,

University of Yangon.
Nutritional values of leaves
Carbohydrate, protein, fibre, fat, vitamin B1 and vitamin C contents
have been determined in National Nutritional Centre, Department of Health,
according to the procedures given in the methods of A.O.A.C. (Horwitz,
1980).
Antimicrobial activity of different solvent extracts of leaves
Extraction and test organisms
The powdered leaves were extracted with chloroform, acetone,
petroleum-ether, methanol, ethyl acetate, ethanol and distilled water by
percolation method. The filtrate solvents were evaporated by using water
bath .
Crude extracts of various solvents were tested on six pathogenic
microorganisms such as Bacillus pumalis, Bacillus subtilis, Escherichia coli,
Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans.
The test organisms used in this research were supplied from the
Development Centre for Pharmaceutical Technology (DCPT) for
determination of antimicrobial activities.
The study of antimicrobial activities was determined by agar-well
diffusion method described by Cruickshank, et al., (1975). Nutrient agar
was prepared and boiled and 20 - 25 ml of the medium was poured into a
test tube and plugged with cotton wool and steriled at 121˚C for 15 minutes
in an autoclave. Then the tubes were cooled down to 30 - 35˚C and poured
into the sterile petridishes and 0.1 - 0.2 ml of the test organisms were also
added into the dishes. They were allowed to set for 2 - 3 hours. Then 10 mm
agar - well was made by the help of sterilized agar-well cutter. After then
about 0.2 ml of sample was introduced into the agar-well and incubated at
37˚C for 24 hours. The inhibition zones which appeared around the agar -
well indicated the presence of antimicrobial activities. The diameter of the
inhibition zones were measured and recorded in mm, the controlled
experiments using solvent only were prepared for the comparison with
leaves plant extracts.
340 Universities Research Journal 2011, Vol. 4. No. 1

Results
Morphological Characters of Sesbania grandiflora L.
Small tree, stems cylindrical pubescent. Leaves alternate,
unipinnately compound, imparipinnate. Inflorescence axillary racemes.
Flower creamy white and dark red. Stamens (2+3), synsepalous. Pistil 1-
carpel, ovary linear-oblongoid (Fig. 1 – 5).

Pauk-pan- phyu Pauk-pan-ni

Fig.(1) Habit Fig.(2) Habit Fig.(3) Leaves Fig.(4) Flowers Fig.(5) Fruits

Qualitative and quantitative analysis of powdered leaves

Qualitative analysis
Qualitative analyses were concerned with determination of presence
or absence of phytochemical in qualitative value. The investigation of these
tests confirmed the presence of alkaloids, α-amino acid, carbohydrate,
starch, reducing sugar, glycoside, tannin, saponin, phenolic compound,
steroid/ terpenoid and flavonoid in Pauk-pan-phyu and Pauk-pan-ni leaves.
Cyanogenic glycoside were absent in the leaves of both plants. The results
were tabulated in Table (1).
 Universities Research Journal 2011, Vol. 4. No. 1 341

Table (1) Qualitative analysis of Sesbania grandiflora L.

Results

Solvent used Pauk- Pauk-

No. Test Test reagent Observation pan- pan-ni
for extraction
phyu
leaves leaves
1 Alkaloids 1%HCl Dragendroff’s Orange ppt + +
reagent
Mayer’s regent White ppt + +
Wagner’s reagent Reddish Brown
Sodium picrate ppt + +
solution Yellow ppt + +
2 α-amino-acid DW Ninhydrin Pink spot + +
reagent
3 Carbohydrate DW 10% α-naphtha Red ring + +
and conc: H2SO4

4 Starch EtOH Iodine solution Blue black ppt + +

5 Reducing sugar Dil H2SO4 Benedict’s Brick red ppt + +

solution
6 Cyanogenic DW Sodium picrate No colour - -
glycoside solution
7 Glycoside EtOH 10% lead acetate Pale yellow ppt + +
solution
8 Phenolic compound EtOH 3% ferric chloride + +
solution blue ppt
9 Saponin DW Distilled water Frothing + +
10 Tannin EtOH 3% ferric chloride Yellow ppt + +
solution

11 Steroid/Terpenoid Pet-ether Acetic anhydride Green ppt + +

and conc: H2SO4

12 Flavonoid EtOH EtOH, Mg, conc: Pink colour + +

HCl
DW=Distilled Water
342 Universities Research Journal 2011, Vol. 4. No. 1

Quantitative analysis of Sesbania grandiflora L.

From the results of quantitative analysis, the moisture content was
usually determined by drying to constant weight and taking the loss in
weight as moisture. Total ash, acid insoluble ash, and water soluble ash
content were also recorded. All these values were useful for the quality
control system regarding ash and it was used for medicinal purposes. The
solubilities of leaves powder plant in acetone, chloroform, petroleum ether,
ethyl acetate, ethanol, methanol and distilled water were carried out to
determine the amount of total solids soluble in an individual solvent.
Sesbania grandiflora L. (Pauk-pan-phyu and Pauk-pan-ni) leaves were
found to be significantly soluble in methanol and ethanol than those of other
solvents. (Table 2)

Table (2) Quantitative analysis of Sesbania grandiflora L.

Content in %
No Physicochemical characters Pauk-pan-phyu Pauk-pan-ni
leaves leaves
1 Moisture content 9.75 9.09
2 Total Ash content 10.15 11.38
3 Petroleum ether soluble content 3.33 3.00
4 Chloroform soluble content 3.40 3.25
5 Ethyl Acetate soluble content 3.35 3.00
6 Acetone soluble content 6.00 6.67
7 Methanol soluble content 33.3 30.0
8 Ethanol soluble content 28.5 28.0
9 Distilled water soluble content 10.05 10.0
10 Acid soluble ash content 2.00 1.82
11 Acid insoluble ash content 3.08 3.20
Universities Research Journal 2011, Vol. 4. No. 1 343

Elemental analysis and nutritional values of Sesbania grandiflora L.

Elemental analysis was to determine the amount of elements in
Sesbania grandiflora L. (Pauk-pan-phyu and Pauk-pan-ni) leaves. It was a
quantitative indication of the level of nutrients in plant obtained by using
EDXRF (Energy Dispersive X-ray Fluorescence) spectrometer. It was
found that Calcium (Ca), Phosphorus (P), Potassium (K), Sulphur(S),
Chlorine (Cl) and Iron (Fe) are macronutrient elements, where as Strontium
(Sr), Brominr (Br), Manganese (Mn) and Zinc (Zn) are found to be as
micronutrient elements. Above the elements were relatively abundant in the
leaves of both samples heavy toxic metals Lead (Pb), Arsenic (As) and
Mercury (Hg) were absent. (Table 3)
Table (3) Comparative studies on relative elements of Sesbania grandiflora
L. by EDXRF
Relative abundance in %
No. Elements Pauk-pan-phyu Pauk-pan-ni
leaves leaves
1 Calcium (Ca)** 1.327 1.072
2 Phosphorus(P)** 0.997 0.700
3 Potassium(K)** 0.447 0.548

4 Sulphur(S)** 0.288 0.203

5 Chlorine(Cl)** 0.225 0.106

6 Iron(Fe)** 0.021 0.022

7 Strontium(Sr)* 0.013 0.008

8 Bromine(Br)* 0.011 0.006

9 Manganese(Mn)* 0.011 0.011

10 Zninc(Zn) 0.003 -
**= macronutrient, *= micronutrient

Elemental analysis by AAS

Due to the results of AAS analysis, Cu content in powdered leaves
of Pauk-pan-phyu and Pauk-pan-ni were 3.65 and 4.71 ppm respectively
which was found to be higher than the other elements. (Table 4)
344 Universities Research Journal 2011, Vol. 4. No. 1

Table (4) Mineral values of Sesbania grandiflora L. by using AAS

Pauk-pan-phyu leaves
No. Mineral elements Pauk-pan-ni leaves (ppm)
(ppm)
1 Copper 3.65 4.71
2 Potassium 0.01 2.08
3 Calcium 0.37 0.56
4 Iron 0.03 0.03
5 Phosphorus 0.05 0.06

Nutritional values of Sesbania grandiflora L.

The nutritional values of powdered plant were carried out at the
National Nutritiional Center, Department of Health and Ministry of Health.
According to the experiments, the amount of carbohydrate and protein were
greater than fiber and fat. Although vitamin B1 was 0.21 mg and 0.19 mg,
and the amount vitamin C 105.74 mg and 120.63 mg were found to be
respectively. (Table 5)
Table (5) Nutritional values of Sesbania grandiflora L.
Pauk-pan- Pauk-pan-ni
No. Type of nutrients
phyu leaves leaves
1 Carbohydrate 32.81 g 21.31 g
2 Fat 6.59 g 9.53 g
3 Fibers 6.02 g 6.94 g
4 Protein 34.68 g 40.75 g
5 Vitamin B1 0.21 mg 0.19 mg
6 Vitamin C 105.74mg 120.63 mg

Antimicrobial activity of different solvent extracts of

Sesbania grandiflora L.
In the experiment, the antimicrobial activities of crude extracts were
carried out by using various solvent such as petroleum ether, ethyl acetate,
distilled water, ethanol, acetone, chloroform and methnol. In this results,
Universities Research Journal 2011, Vol. 4. No. 1 345

among various extracts of leaves Sesbania grandiflora L. (pauk-pan-phyu

and pauk-pan-ni), chloroform extract showed the highest activity on all six
different types of microorganisms especially on Pseudomonas aeruginosa.
The results were shown in Figure (6 -11).

Discussion and Conclusion

In this research, chemical studies such as preliminary phytochemical
investigation, physicochemical characterization, elemental analysis,
examination of nutritional values and vitamins contents of Sesbania
grandiflora L. (Pauk-pan-phyu and Pauk-pan-ni) leaves had been studied.
For preliminary phytochemical investigation, Sesbania grandiflora
L. (Pauk-pan-phyu and Pauk-pan-ni) leaves plant samples were collected
from Hpa-an and prepared, alkaloids, α-amino acid, carbohydrate, starch,
reducing sugar, glycoside, tannin, saponin, phenolic compound,
steroid/terpenoid and flavonoid were detected in leaves. Cyanogenic
glycoside was absent in the leaves of both plants. Therefore, the plants
Sesbania grandiflora L. (Pauk-pan-phyu and Pauk-pan-ni) have
phytoconstituents. Physicochemical characteristics such as moisture
contents, ash content and different solubility of organic constituents in
organic solvents were determined. The solubility in methanol and ethanol
were found to be the greatest in these solubility tests. The more polar
phytoconstituents can also be found.
According to the EDXRF result of elemental analysis, it was found
that the macronutrient elements in calcium, phosphorus, potassium, sulphur,
chlorine and iron. The micronutrient elements in strontium, bromine,
manganese and zinc were found in Pauk-pan-phyu and Pauk-pan-ni leaves.
346 Universities Research Journal 2011, Vol. 4. No. 1

Antimicrobial activity of different solvent extracts of

Sesbania grandiflora L. (Pauk-pan-phyu and Pauk-pan-ni) leaves

Pauk -pan -
No Solvent Pauk -pan -ni
phyu

1 Petroleum - 11 mm 10 mm
ether Ba

2 Chloroform 15 mm 20 mm

3 Methanol 12 mm 13 mm

4 Acetone 10 mm 11 mm Ba

5 Ethyl acetate 11 mm 10 mm
Table (6) Organisms -
6 Ethanol 15 mm 15 mm Bacillus pumalis
7 Distilled water 12 mm 8 mm Fig .( 6 )

Agar-well = 7mm
Table (7) Organisms - Bacillus subtilis
Pauk -pan - Pauk -pan -
No Solvent
phyu ni

1 Petroleum - 12 mm 9 mm
ether Pauk-pan-ni

Pauk –pan-phyu
2 Chloroform 16 mm 20 mm

3 Methanol 8 mm -

4 Acetone 8 mm - Pauk-pan-ni

5 Ethyl acetate 10 mm 9 mm Fig.(7)

6 Ethanol 14 mm 10 mm
Agar-well = 7mm
7 Distilled water 13 mm -
Universities Research Journal 2011, Vol. 4. No. 1 347

Table (8) Organisms - Escherichia coli

Pauk -pan - Pauk -pan -

No Solvent
phyu ni

1 Petroleum -ether 11 mm 11 mm

2 Chloroform 15 mm 20 mm

3 Methanol 12 mm 14 mm

4 Acetone 11 mm 15 mm

5 Ethyl acetate 10 mm 10 mm Fig.(8)

6 Ethanol 14 mm 16 mm

7 Distilled water 10 mm 8 mm

Agar-well = 7mm
Table (9) Organisms – Pseudomonas aeruginosa
Pauk -pan Pauk -pan -
No. Solvent
-phyu ni

1 Petroleum -ether 10 mm 12 mm

Pauk –pan-phyu
2 Chloroform 17 mm 21 mm

3 Methanol 12 mm 14 mm

4 Acetone 10 mm 15 mm
Pauk-pan-ni

5 Ethyl acetate 9 mm 10 mm Fig.(9)

6 Ethanol 12 mm 15 mm

7 Distilled water 10 mm 12 mm

Agar-well = 7mm
348 Universities Research Journal 2011, Vol. 4. No. 1

Table (10) Organisms- Staphylococcus aureus

Pauk -pan - Pauk -pan -

No. Solvent
phyu ni

1 Petroleum -ether 10 mm 9 mm

Pauk –pan-phyu
2 Chloroform 15 mm 19 mm

3 Methanol 12 mm 12mm

4 Acetone 10 mm 12 mm
Pauk-pan-ni
5 Ethyl acetate 10 mm 10 mm
Fig.(10)
6 Ethanol 14 mm 17 mm

7 Distilled water 10 mm -

Agar-well = 7mm
Table (11) Organisms - Candida albicans

Pauk -pan Pauk -pan -

No. Solvent
-phyu ni

1 Petroleum -ether 8 mm 10 mm

2 Chloroform 19 mm 20 mm

3 Methanol 14 mm 13 mm
Pauk –pan-phyu
4 Acetone 13 mm 13 mm

5 Ethyl acetate 8 mm 10 mm

Pauk-pan-ni
6 Ethanol 13 mm 17 mm
Fig.(11)
7 Distilled water 8 mm 9 mm

Agar-well = 7mm
Universities Research Journal 2011, Vol. 4. No. 1 349

There are six main classes of nutrients that the body needs:
carbohydrates, proteins, fats, vitamins, minerals, and water. It is important
to consume these six nutrients on a daily basis to build and maintain healthy
bodily function. The important nutritional values found in this plant might
also be major contributors to the medicinal value of Pauk-pan-phyu and
Pauk-pan-ni.
The antimicrobial activities with six different microorganisms were
also tested by using seven crude extracts such as chloroform, acetone,
petroleum ether, methanol, ethanol, aqueous extract of Pauk-pan-phyu and
Pauk-pan-ni leaves. In this experiment, chloroform extracts of Pauk-pan-
phyu and Pauk-pan-ni leaves showed the highest activity on test organisms.
Methanol, acetone and aqueous extracts were activity of Bacillus subtilis
and aqueous extracts was no activity of Staphylococcus aureus Pauk-pan-ni
leaves.
From the overall assessment of chemical studies of Pauk-pan-phyu
and Pauk-pan-ni plants which can be used as nutritious food due to the
results of the vitamins and nutrient elements. Furthermore, the presence of
phytoconstituents, steroids, and flavones in these plants indicate the
medicinal properties.

Acknowledgements
I would like to express my deepest gratitude and special thanks to Dr. Thet Thet
May, Professor and Head and Professor Dr. Aye Pe, from the Department of Botany,
University of Yangon for their invaluable advice and kind suggestions. I would like to
express my deepest gratitude to my supervisor Dr. Moe Moe Lwin, Lecturer, Department
of Botany, Pathein University for her close supervision, valuable guidance, constant
encouragement for the success of my research work..

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