International Journal of Research in Engineering and Bioscience ISSN: 2321-743X

Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF

TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR

Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A

Department of Botany, St. Joseph’s College (Autonomous), Thrichirappali-620 002, Tamil

Nadu, India

ABSTRACT
India is endowed with a rich wealth of medicinal plants. Some of these
plants are the potential source of antibiotics for various infections. The
antimicrobial activity of Tragia involucrata L. stem and leaf extracts was determined
against 7 pathogenic bacteria and fungi by disc diffusion method. The Chloroform
stem extract and Ethanol leaf extract has showed higher inhibition in bacterias like
Pseudomonas aeruginosa and Vibrio cholerae. Ethanol stem extract and Methanol
stem extract has more effect against fungi like Aspergillus niger and Rhizopus
arrhizus. There was no activity against Penicillium chrysogenum. Preliminary
phytochemical screening was carried out for qualitative determination of plant
extracts. The secondary metabolites such as Alkaloids, Flavonoids, Steroids,
Lignin’s, Quinines and Triterpenoides. The different peaks were obtained in the UV-
VIS profile and by the FTIR, it was confirmed that the presence of Alcohol, Amides,
Phosphines, Anhydrides, Carboxylic acids, Alkenes, Sulfoxides and Alkyl halides.

Keywords: Tragia involucrata L, Antimicrobial activity, Phytochemical analyzes, UV-

Vis, FTIR.

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 82
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

INTRODUCTION important in the production of

Medicinal plants constitute an economically important organic
important natural wealth of a country. compounds and pharmaceuticals. The
India is endowed with a rich wealth of rising incidence of multidrug resistance
medicinal plants. Plants have been an amongst pathogenic microbes has further
essential part of human society since the necessitated the need to search for newer
start of civilization. Medicinal plants are a antibiotic sources. The use of plant
boon of nature to cure a number of extracts and Phytochemicals with known
ailments of human beings. Ayurveda and antimicrobial properties can be of great
Unani systems of medicine regularly significance in therapeutic treatments
employ a large number of Indian (Sahaya Sathish et al., 2011).
medicinal plants as antibiotic agents. Our
In recent years attempts are being
country represents a store house of genetic
made to screen out the efficiency of better
diversity of plants (Perumal et al., 2004).
known plants which have medicinal
In India around 20,000 medicinal plants
values, especially to find out their
have been recorded recently, but more than
antimicrobial properties against different
500 traditional communities use about 800
pathogenic microbes. Several countries
plant species for curing different diseases
have already done analytical studies on
(Kalaivani et al., 2012).
better known medicinal plants for
Uses of antimicrobial agents controlling diseases with this motive. The
against harmful pathogen are known to screening of plants extracts and natural
have been common practice at least 2000 products for antimicrobial activity has
years. In order to promote the use of revealed the potential of higher plants as a
medicinal plants as potential sources of source of the new anti-infective agents as
antimicrobial compounds, it is important well as serving drug discovered from
to thoroughly investigate their composition natural products for primary and secondary
and activity and thus validate their use compounds. Antimicrobial agents include
(Nair et al., 2006). Some Phytochemicals all classes of secondary metabolites. These
produced by plants have antimicrobial principles play an important role in the
activity and used for the development of biochemical resistance against pathogenic
new antimicrobial drugs (Sahaya Sathish organisms. Some of these plants are the
et al., 2012). Higher plants are much more potential source of antibiotics for various

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 83
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

infections. Keeping all this in mind, in the obtained collected from Govt. Medical
present investigation an attempt was made College, Tiruchirappalli, Tamil nadu,
in the medicinal plant to study the India.
antimicrobial activity and phytochemical
Antibacterial activity assay
constituents of T. involucrata L. using
The antibacterial activity of the
UV-VIS, FTIR.
plant extracts were tested by the modified
MATERIAL AND METHODS disc diffusion method (Baur et al., 1966).
Collection and Preparation of different The bacterial inoculums (20 h broth) were
Plant extracts uniformly spread over the agar plates
Healthy, disease free and mature using a glass L-rod. A total of 0.2ml of
plant collected from Madurai. Madurai each extract was aseptically added to the
district, Tamil Nadu, India. Successive discs (0.5mm diameter) and allowed to dry
solvent extraction-the T. invocrata L. plant before being placed on the top of the agar
were taken and subjected to successive plate. The plates were incubated at 37°C
solvent extraction. The plant leaves and for 24h and the diameter of growth
stem were collected, washed with distilled inhibition zone was recorded. A standard
water and dried under shed. The dried antibiotic for Bacteria, Chloramphenicol
samples were grinded to fine powder. 30 and Fungi, Gentamicine was used as
gm of powdered sample was mixed with positive controls.
50 ml of solvent (Ethanol, Methanol,
Phytochemical analysis
Chloroform and Aqueous) kept
The Ethanol leaf extract was
undisturbed for 3 days. Finally the solvents
selected for Preliminary Phytochemical
were filtered to collect the plant extract.
screening because of their better
Tested Microorganisms antimicrobial activity. Compounds
Tested Microorganisms were identification by chemical tests. Tests for
Bacillus subtilis, (Gram positive bacteria), Steroids, Triterpenoides, Glycosides,
Pseudomonas aeruginosa, Salmonella Carbohydrate, Alkaloids, Phenolic
typhi, Vibrio cholerae (Gram negative compound, Catachins, Flavanoids,
bacteria) and Penicillium chrysogenum, Saponins, and Tannins were carried out
Aspergillum niger, Rhizopus arrhizus according to the methods of (Brindha et
(Fungus). The Microorganisms were al., 1981).

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 84
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

UV-VIS and FTIR Spectroscopic Chloroform stem extract showed the

analysis higher inhibition zone (1.26cm) in P.
The extracts were examined under aeruginosa, B. subtilis, V. cholerae,
visible and UV light for proximate and R. arrhizus (Table-2). Hence the
analysis For UV-VIS and FTIR antimicrobial activity of various solvent
spectrophotometer analysis, the extracts extracts of leaf and stem on selected
were centrifuged at 3000 rpm for 10 min microorganisms clearly revealed that the
and filtered through Whitman No. 1 filter aqueous extract was found to possess
paper by using high pressure vacuum significant inhibitory property. Both
pump. The sample is diluted to 1:10 with aqueous extracts showed the less inhibition
the same solvent. The extracts were on pathogenic strains (0.7cm). The
scanned in the wavelength ranging from antimicrobial activity was maximum of
300-1100 nm using Perkin Elmer Ethanolic extract while minimum activity
Spectrophotometer and the characteristic is in Aqueous extract (Table-1 & 2).
peaks were detected. FTIR analysis was However P. chrysogenum are resistance to
performed using Perkin Elmer all the solvent extracts.
Spectrophotometer system, which was
The qualitative UV-VIS spectrum
used to detect the characteristic peaks and
profile of T. invocrata L. Ethanolic stem
their functional groups. The peak values of
extract was selected at wavelength from
the UV-VIS and FTIR were recorded.
350 to 1100 nm due to sharpness of the
Each and every analysis was repeated
Ethanolic peaks and proper baseline. The
twice for the spectrum confirmation.
profile showed the peaks at 350 and
RESULTS AND DISCUSSION 975nm with the absorption of 353.68 and
In Ethanolic leaf extract, the 974.14 respectively (Fig. 1; Table- 5) and
antimicrobial activity in P. aeruginosa, B. UV-VIS profile of Ethanolic leaf extract of
subtilis, V. cholerae, A. niger, R. arrhizus T. invocrata L. was chosen at a
showed higher inhibition zone (1.63cm, wavelength of 400 to 1100 nm and the
Table-1). The inhibitory action was profile showed the peaks at 423.26, 617.64
observed in terms of inhibition zone and 676.47nm with the absorption 425.52,
formed around each disc caused by the 616.18 and 674.77 respectively (Fig. 2;
diffusion of antimicrobial substances from Table-5). Performing the next advanced
the paper disc into the surrounding media. phytochemical analysis technique of FTIR,

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 85
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

the presence of various functional group of used for the identification of chemical
different compound was found. Every constituents present in T. invocrata L
solvent had its respective functional group (Fig.3; Table-4). In addition, UV-VIS and
like Alcohol, Amides, Phosphines, FTIR spectroscopy is proved to be a
Carboxylic acids, Alkenes, Sulfoxides, reliable and sensitive method for detection
Alkyl halides. Hence, the crude extracts of biomolecular composition. (Komal
subjected to UV-VIS and FTIR analysis is Kumar et al., 2011).

Table: 1. Antimicrobial activity of leaf extract of T. involucrata L.

Zone of Inhibition (cm)*
Selected Microbes Ethanol Methanol Chlorofor Aqueous Standard
(Bacteria & Fungi) extract extract m extract extract antibiotics
P. aeruginosa 1.2±0.28 1.26±0.15 0.76±0.15 _ 2
B. subtilis 1.0±0.95 0.86±0.75 0.56±0.51 0.53±.0.47 2.1
S. typhi _ _ 0.93±0.32 _ 2
V. cholerae 1.63±0.20 1.26±0.05 1.16±0.25 _ 2
P. chrysogenum _ _ _ _ 1.9
A. niger 0.86±0.30 0.86±0.30 _ 0.83±0.15 2
R. arrhizus 1.5±0.20 1.3±0.20 0.96±0.56 0.73±0.20 2.1
Note: *values are the mean of triplicate, Standard antibiotics (Bacteria Chloramphenicol,
Fungi Gentamicine), ± standard deviation

Table: 2. Antimicrobial activity of stem extract of T. involucrata L.

Zone of Inhibition (cm)*
Selected Microbes Ethanol Methanol Chlorofor Aqueous Standard
(Bacteria & Fungi) extract extract m extract extract antibiotic
s
P. aeruginosa 1.06±0.45 0.95±0.15 1.3±0.26 0.8±0.43 1.9
B. subtilis _ _ 1.03±0.55 0.7±.0.35 2.1
S. typhi 0.83±0.46 1.16±0.46 0.93±0.35 _ 2
V. cholerae _ _ 0.93±0.55 0.83±0.11 1.9
P. chrysogenum _ _ _ _ 1.9
A. niger 0.8±0.3 0.8±0.3 _ 0.63±0.15 2
R. arrhizus 1.13±0.20 1.26±0.20 0.96±0.56 0.66±0.20 1.9
Note: *values are the mean of triplicate, Standard antibiotics (Bacteria Chloramphenicol,
Fungi Gentamicine), ± standard deviation

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 86
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

Table: 3. Phytochemicals screenings of T. involucrata L. Ethanol leaf extract.

S. No Phytochemicals Ethanol leaf extract

1 Steroids +
2 Triterpenoides +
3 Glycosides +
4 Carbohydrate +
5 Alkaloids +
6 Phenolic compound -
7 Catachins -
8 Flavanoids +
9 Saponins -
10 Tannins -
Note: + represent as present; - represents as absent

Table: 4. FTIR peak values of Ethanol leaf extracts of T. involucrata L.

Function group Molecular motion Wave number (cm-1)
Alcohol O-H stretch ~3696.72
Amides N-H stretch ~3434.77
Phosphines P-H stretch ~2355.95
Anhydrides C=O stretch 1813.91
Carboxylic acids C=O stretch 1613.87
Alkenes C=C stretch (conjugated) 1613.87
Alkenes C-H in –plane bent 1404.51
Sulfoxides S=O stretch 1030.40
Alkyl halides C-CI stretch 776.33
Alkynes Acetylenic C-H bent 685.24

Table 5: UV-VIS peak values of Ethanolic stem & leaf extracts of T. involucrata L.
S. No Ethanolic stem extract Ethanolic leaf extract
nm Absorption nm Absorption
1. 350.00 353.69 425.52 423.26
2. 506.25 505.22 617.64 616.18
3. 525.00 535.89 676.47 674.77
4. 606.25 605.86 __ __
5. 656.25 661.94 __ __
6. 975.00 974.14 __ __

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 87
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

Fig. 1: UV-VIS Spectrum of Ethanolic stem extracts of T. involucrata L

Fig. 2: UV-VIS Spectrum of Ethanolic leaf extracts of T. involucrata L.

Fig. 3: FTIR Spectrum of Ethanolic leaf extracts of T. involucrata L.

ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 88
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

SUMMARY AND CONCLUSION analysis of T. involucrata L is to be

The phytochemical analysis and concluded that this plant have bioactive
antimicrobial activity was performed in the compounds like Steroids, Triterpenoides,
leaf and stem of T. involucrata L. the Glycosides, Carbohydrate, Alkaloids,
antimicrobial activity was studied by disc Phenolic compound, Catachins,
diffusion method in the 3 different solvent Flavanoids, Saponins, and Tannins form
extract and Aqueous extract of the plant T. the characteristic nature for its medicinal
involucrata L. leaf and stem. There was uses. These compounds also possess
antimicrobial activity observed in the all antimicrobial activity against pathogenic
the 4 tested bacterial and 2 fungal species. microbes as well. This Indian herb serves
The Chloroform stem extract and Ethanol to cure many diseases an also used for
leaf extract has showed higher inhibition various other herbal treatment.
in P. aeruginosa (1.3cm) and V. cholera
ACKNOWLEDGEMENT
(1.6cm). Ethanol stem extract and
I heartily thankful to Arch Bishop
Methanol stem extract has more effect
Casimier Instrumentation center, St.
against A. niger (1.5cm) and R. arrhizus
Joseph’s College (Autonomous),
(1.26cm). There was no activity against P.
Tiruchirappalli, Tamil Nadu, India for
chrysogenum. On taking leaf extract,
providing the instrumentation facilities for
preliminary phytochemical analysis
UV-Vis and FTIR analysis.
(Brindha et al., 1981) was performed
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ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 89
 International Journal of Research in Engineering and Bioscience ISSN: 2321-743X
Special Issue: National conference on “Drug discovery, Threatened medicinal plants and
strategies for sustainable use” EDITED BY, S. DOMINIC RAJKUMAR & J. K. LAL
St. Andrew’s college, Gorakhpur, U.P., India

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ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF TRAGIA INVOLUCRATA L. USING UV-VIS AND FTIR –
Sahaya Sathish S, Vijayakanth P, Palani R, Thamizharasi T and Vimala A 2013 Vol. 1 (Special Issue 1) Page 90