Indo American Journal of Pharmaceutical Research, 2015 ISSN NO: 2231-6876

SPECTRAL AND ANTIBACTERIAL STUDIES ON SOME ETHNOBOTANICALLY

IMPORTANT MEDICINAL PLANTS USED BY THOTTIANAICKANS OF SEMMALAI
Thiravia Doss Dons, Sebastian Soosai Raj *
Department of Botany, St. Joseph’s College, Tiruchirappalli – 620002, Tamilnadu, India.

ARTICLE INFO ABSTRACT

Article history Ethanolic extract from four medicinal plants, Justicia tranquebariensis, Tribullus terrestris,
Received 27/07/2015 Aloe vera and Curcuma longa were tested for their antibacterial activity against four wound
Available online associated infectious microorganisms such as Staphylococcus aureus, Bacillus subtilis, E. coli
31/07/2015 and Klebsiella pneumonia by disc diffusion method. The Ethanolic stem extracts of Justicia
tranquebariensis showed significant antibacterial activity against all test organisms. The
Keywords spectrum of activity observed in the present study may be indicative of possible source of
Disc Diffusion Method, new and effective drugs from these herbals. The Ethanolic extracts of these four species were
Antibacterial Activity, analyzed to identify electron transition and the functional group by FT–IR and UV
FT –IR And UV – Spectroscopy. The presence of the secondary alcohol, phenol, amines, alkanes and halogen
Visible Spectroscopy has been revealed by FT-IR analysis. UV –VIS analysis of Justicia tranquebariensis,
Analysis. Tribullus terrestris and Curcuma longa confirm the presence of Aniline.

Corresponding author
Dr. S. Soosai Raj
M. Sc., Ph. D,
Assistant Professor,
Department of Botany
St. Joseph’s College
Tiruchirappalli - 620 002
[email protected]
9750124125

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Please cite this article in press as Thiravia Doss Dons et al. Spectral and Antibacterial studies on some ethnobotanically
important medicinal plants used by Thottianaickans of Semmalai. Indo American Journal of Pharm Research.2015:5(07).

Copy right © 2015 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Page

Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Vol 5, Issue 07, 2015. Thiravia Doss Dons et al. ISSN NO: 2231-6876

INTRODUCTION
Plants are rich in nutrients thus they serve as source of food, besides plants are also rich in phyto compounds which have
healing abilities. From earliest times, even when we were lacking the knowledge about the compounds present in the plants and their
mode of action, plants were used for the treatment of diseases. Over the centuries, societies around the world have developed their
own tradition to make sense of medicinal plants and their uses. The wide spread use of herbal remedies and health care preparations
elevated from ordinarily used traditional herbs to the occurrence of natural products with medicinal properties [1].
More recently, a lot of emphasis has been paid to use eco – friendly products that are having a fewer side effects. According to the
records, 80% of the world population have loyalty in traditional medicine, especially plant – based drugs for their primary health care
[2].
Scientific investigations of medicinal plants have been initiated in many countries because of their contributions to health
care. The primary benefits of using plant – derived medicines are relatively safer than synthetic alternatives, offering profound
therapeutic benefits and more affordable treatment [3].
There has been an increasing incidence of multiple resistances in human pathogenic microorganisms in recent years, largely
due to indiscriminate use of commercial antimicrobial drugs commonly employed in the treatment of infectious diseases. Numerous
studies were undertaken to identify compounds that are plants based effective as antibiotics [4]. Traditional healing systems around
the world that utilize herbal remedies are an important source for the discovery of newer antibiotics [5]. Some traditional remedies
have already produced compounds that are effective against antibiotic – resistant strains of bacteria [6]. The results indicate the need
for further research into traditional health systems [7]. It also facilitates pharmacological studies leading to synthesis of more potent
drug with reduced toxicity [8]. The need of the hour is to screen a number of medicinal plants for promising biological activity. In the
present work four different medicinal plants were evaluated for their antibacterial potency. The purpose of this study is to evaluate the
FT-IR, UV-vis and antibacterial effect of few selected species based on their traditional use by the local people.

MATERIALS AND METHODS

Semmalai
Semmalai hills, the area of investigation, lies within Tiruchirappalli district, just next to Dindigal and Karur district
boundaries approximately within 10°.10' to 11°.20' North latitude and 77°.40' to 78°.60' East longitudes. At an elevation ranging from
300-600m above sea level. During the months of October – November, the district receives a rainfall. Temperatures for the district
have been 25°-30°C respectively. The soil is mostly red and loamy.
Thottianaickans live in Ondibommanayakkanpatty, Desirnayakkanpatty, Kandamanayakkanur, Valiampatty,
Palattanayakkanur are the areas of their inhabitations in Semmalai hills. Thottianaickans are also referred to as Vaettainayakkans as
their main profession was once hunting, and also as kambalaththunayakkars as they always cover their body with a rug made of
animal hairs (Kambalam). Their name is supposed to be derived from Thottil; because each one carries a Thottil (cradle) made of cloth
or rug on their shoulder, either to carry a baby or a produce from the forest. Their diet includes meat of rats, cats, frogs, snails,
squirrels, birds, wild pigs etc. Their language is a corrupt form of Telugu and Tamil [9].

Collection of medicinal plants

Justicia tranquebariensis, Tribullus terrestris, Aloe vera and Curcuma longa were selected based on their ethno medical
importance. Healthy disease free leaves, stems of selected plants were collected in and around Semmalai, Karur district, shade dried,
pulverized and stored at 4°C. until further use.

Preparation of Plant extracts

Each plant powder (10g) was taken as equimolar ratio in 30ml of ethanol, totally 40g / 120ml in a single sterile bottle and left
undisturbed for 48hrs at 4°C. Thereafter, it was filtered through muslin cloth and the extract was collected and the solvent was
evaporated and the final weight is measured. Then, the extract was stored under refrigeration for further studies.

Test microorganisms
Staphylococcus aureus, Bacillus subtilis, E. coli and Klebsiella pneumonia.

Preparation of sterile disc

Whatman No. 1 filter paper was used to prepare disc of 6 mm diameter.

Antimicrobial study
Assay of antibacterial activity using Disc diffusion method
The antibacterial activity of selected plants was evaluated by agar disc diffusion method. The microorganisms was activated
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by inoculating a loopful of inoculums in the nutrient broth medium and incubated for 24hrs. 20ml of sterilized nutrient agar was
poured into sterile petriplates, after solidification, fresh culture of human pathogens was swabbed on the respective plates.
Disc of 6 mm diameter was impregnated with plant extract was placed on the medium. Disc soaked in pure solvents served as negative
control and antibiotic (Chloramphenicol) discs were used as positive control. The plates were then incubated at 37°C for 24h. The
antibacterial activity measured in terms of zone on inhibition (mm) of microbial growth after incubation.
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Vol 5, Issue 07, 2015. Thiravia Doss Dons et al. ISSN NO: 2231-6876

UV-VIS and FT-IR Spectroscopic analysis

The extracts were examined under visible and UV light for proximate analysis. For UV-VIS and FTIR spectrophotometer
analysis, the extracts were centrifuged at 3000 rpm for 10 min and filtered through Whatmann No. 1 filter paper by using high
pressure vacuum pump. The extracts were scanned in the wavelength ranging from 200-1100 nm using Perkin Elmer
Spectrophotometer and the characteristic peaks were detected. FTIR analysis was performed using Perkin Elmer Spectrophotometer
system, which was used to detect the characteristic peaks and their functional groups. The peak values of the UV-VIS and FTIR were
recorded. Each and every analysis was repeated twice for the spectrum confirmation [10].

RESULTS AND DISCUSSION

Antibacterial activity was conducted against gram positive (Staphylococcus aureus, Bacillus subtilis) and gram negative
bacteria (E. coli, Klebsiella pneumonia). The result was compared with that of the standard Chloramphenicol (30µg) antibacterial drug
and the results are tabulated Table I, Figure I.

Table I : Antibacterial activity of ethanol extracts of selected plants and antibiotic against some human pathogenic bacteria
(Zone of inhibition measured in mm).

Test organism Plant extracts and their zones Chloramphenicol +ve Ethanol –ve control
of inhibition (mm) Control (mm) (mm)
Staphylococcus aureus 14 37 -
Bacillus subtilis 18 19 -
E. coli 16 29 -
Klebsiella pneumonia 15 16 -

Figure I: Antibacterial activity of Ethanol extract of medicinal plants against human pathogenic bacteria using Disc diffusion
method.

In our study among the four pathogens a maximum inhibition zone (18mm) was found against Bacillus subtilis, and E. coli
and the moderate inhibition zone (16mm) was recorded against Klebsiella pneumonia. The minimum inhibition zone (14mm) was
recorded against Staphylococcus aureus.
The FT-IR and UV spectrum was used to identify the functional group of the active components based on the peak value in
the region of infrared radiation [11]. The results of FT-IR analysis of Justicia tranquebariensis was confirmed the presence of the
secondary alcohol, phenol, silicon, aldehydes, ketones, alkanes and lower to halogen, ethers, alkynes. Stretching which shows peaks
range between 659.88 to 3492.49 cm-1 Table II, Figure II.
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Vol 5, Issue 07, 2015. Thiravia Doss Dons et al. ISSN NO: 2231-6876

Table II: FT-IR Peak value and its functional groups of stem extracts of Justicia tranquebariensis.

S. No Peak Area Mode of Vibration Functional groups

1. 3392.49 O-H Stretching Alcohol / Phenol
2. 2976.11 CH3, CH2, and CH Stretch Alkanes
3. 2134.25 -N=C=O Isocyanides
4. 1923.54 Si - OR Silicon
5. 1647.26 N-H bond 1 amines
6. 1449.65 CH2 Aldehydes and ketones
7. 1329.37 Ar – NO2 Stretching Nitro
8. 1273.78 C – F Stretching Halogen
9. 1085.22 Si - OR Silicon
10. 1048.12 C – O Stretching Ethers
11. 659.88 C – H Stretching Alkynes

ACIC
St.Joseph's College ( Autonomous)
Trichy-2
Spectrum Name: IR-Bot1.sp

100.0
95
90 2134.25 1923.54

85 437.53
80
1273.78
75
1329.37
70
659.88
65 1647.26 1449.65 880.39
1406.89
60
55
50 2897.86 1085.22
%T
45

40 2976.11
1048.12
35
30
25 3392.49

20
15
10

5
0.0
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
cm-1

Figure II: FT-IR Peak value and its functional groups of Justicia tranquebariensis.

Tribullus terrestris the functional groups of the components were separated based on its peak such as secondary alcohol,
phenol, amines, alkanes, carboxylic acid, ammonium ions, silicon, halogen, alkynes and ethers which shows peaks range between
660.70 to 3399.81cm-1 Table III, Figure III.

Table III: FT-IR Peak value and its functional groups of stem with leaf extracts of Tribullus terrestris.

S. No Peak Area Mode of Vibration Functional groups

1. 3399.81 O – H Stretching Alcohol / Phenol
2. 2976.36 CH3, CH2, and CH Stretching Alkanes
3. 2898.92 CH3, CH2, and CH Stretching Alkanes
4. 2131.96 -N=C=O Isocyantes
5. 1923.59 SI - OR Silicon
6. 1405.68 CH2 bending-stretching Aldehydes and ketones
7. 1333.67 C – O Stretching Alcohols
8. 1273.30 C – F Stretching Halogen
9. 1084.95 Si - OR Silicon
10. 1048.59 C – O Primary Alcohols
11. 880.61 C – H Stretching Alkanes
12. 660.70 C –CL Stretching Halogen
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Vol 5, Issue 07, 2015. Thiravia Doss Dons et al. ISSN NO: 2231-6876

ACIC
St.Joseph's College ( Autonomous)
Trichy-2
Spectrum Name: IR-Bot2.sp

100.0
95
90
2131.96 1923.59
85
438.57
80 1273.30
75 1333.67
660.70
70
880.61
65 1647.31
1405.68
60
55 2898.92
1084.95
50
%T
45 2976.36
1048.59
40
35
30
25 3399.81

20
15
10

5
0.0
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
cm-1

Figure III: FT-IR Peak value and its functional groups of Tribullus terrestris.

In Aloe vera the functional groups of alcohol, phenol, ammonium ions, carboxylic acid, silicon, and lower to halogen, ethers,
and alkynes was observed with peaks range between 661.56 to 3397.87cm-1 Table IV, Figure IV.

Table IV: FT-IR Peak value and its functional groups of whole plant extracts of Aloe vera.

S. No Peak Area Mode of Vibration Functional groups

1. 3397.87 O – H Stretching Alcohol / Phenol
2. 2946.97 N–H Ammonium ions
3. 2836.28 C – O – CH3 Ethers
4. 2525.01 N–H Ammonium ions
5. 2364.52 P–H Phosphine
6. 2210.43 -N = C = O Isocyantes
7. 1812.35 Si - OR Silicon
8. 1599.59 N – H def Amides
9. 1445.96 - CH3 def Alkanes
10. 1407.70 C – O Stretching Phenols
11. 1114.17 C – F Stretching Halogen
12. 661.56 C – H Stretching Alkynes

ACIC
St.Joseph's College ( Autonomous)
Trichy-2

Spectrum Name: SJ C-Bot-3.s p

100.0
3882.87
95

90 3914.93 1812.35
2364.52
85 3780.63 2210.43
2046.30
80
2525.01
75 1114.17
70

65 1445.96 661.56
1599.59
60 1407.70

55

50
%T 2836.28
45

40
2946.97
35

30

25
1025.87
20
3397.87
15

10

0.0
4000.0 3000 2000 1500 1000 500 400.0
cm-1

Figure IV: FT-IR Peak value and its functional groups of Aloe vera.
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Curcuma longa the functional groups of alkanes, ammonium ions, carboxylic acid, isocyantes, amides, lactams, ethers, and
halogen was found with peaks range between 657.53 to 3370.57cm-1 Table V, Figure V.
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Vol 5, Issue 07, 2015. Thiravia Doss Dons et al. ISSN NO: 2231-6876

Table V: FT-IR Peak value and its functional groups of Rhizome extracts of Curcuma longa.

S. No Peak Area Mode of Vibration Functional groups

1. 2946.82 CH3, CH2, and CH Stretching Alkanes
2. 2835.07 N- H ammonium ions
3. 2525.72 O – H Stretching Carboxylic acid
4. 2363.56 P–H Phosphine
5. 2216.75 -N=C=O Isocyanates
6. 1599.53 N – H def Amides and Lactams
7. 1408.50 -CH2 bending - stretching Aldehydes and Ketones
8. 1113.81 C – H Stretching Alkanes
9. 1025.47 C = C – O – C Stretching Ethers
10. 657.53 C – CL Stretching Halogen

ACIC
St.Joseph's College ( Autonomous)
Trichy-2

Spectrum Name: SJ C-Bot-4.s p

100.0

95

90 3770.57 2363.56 2044.31

85
2216.75
80 1113.81
2525.72
75 1599.53 657.53
70
1408.50
65

60

55 2835.07

50
%T
45 2946.82

40

35 3371.74 1025.47

30

25

20

15

10

0.0
4000.0 3000 2000 1500 1000 500 400.0
cm-1

Figure V: FT-IR Peak value and its functional groups of Curcuma longa.

Hence, the crude extracts subjected to UV- VIS analysis is used for the identification of chemical constituents present in all
four pants. In addition, UV-VIS and FTIR spectroscopy is proved to be a reliable and sensitive method for detection of biomolecular
composition [12].
The qualitative UV-VIS spectrum profile of Justicia tranquebariensis, ethanol extract was selected from, 400 – 1000nm due
to sharpness of peaks and proper baseline. The solvent had its respective functional group like Aniline, Thiobenzopheno and
nitrosobutane Table VI, Figure VI.

Table VI: UV-Spectrum peak value and its functional groups of Justicia tranquebariensis.

S. No Peak Area Functional groups

1. 467.27 Aniline
2. 533.50 Thio benzophenone
3. 611.89 Alkenes, alkynes, aromatics
4. 664.20 Nitrosobutane

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Vol 5, Issue 07, 2015. Thiravia Doss Dons et al. ISSN NO: 2231-6876

Figure VI: UV Spectrum of Ethanol extract of Justicia tranquebariensis.

Tribullus terrestris Ethanolic extract was selected at wavelength from 260 to 700 nm and the profile showed the peaks with
the functional groups of Thiophene, 1-phenyl -1,3-butadiene, nitrosobutane, and isoquinoline Table VII, Figure VII.

Table VII: UV-Spectrum peak value and its functional groups of Tribullus terrestris.

S. No Peak Area Functional groups

1. 264.88 Thiophene
2. 268.98 1-Phenyl -1,3-butadiene
3. 301.02 Nitrosobutane
4. 314.87 Isoquinoline
5. 411.48 Β-carotene
6. 467.94 Thioester
7. 535.67 Tetraphenyl cyclopentadienone
9. 663.96 Nitrosobutane

Figure VII: UV Spectrum of Ethanol extracts of Tribullus terrestris.

The qualitative UV-VIS profile of ethanol extract of Aloe vera was taken at the wavelength of 320 – 700nm and the profile
showed at functional groups of n-Butyl nitrite and nitrosobutane Table – VIII, Figure VIII.

Table VIII: UV-Spectrum peak value and its functional groups of Aloe vera.
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S. No Peak Area Functional groups

1. 363.01 n- Butyl nitrite
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2. 371.04 Jacobsen catalyst

3. 604.12 Anthracene
4. 665.34 Nitrosobutane

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Figure VIII: UV Spectrum of Ethanol extracts of Aloe vera.

The UV – VIS profile Table IX, Figure IX of the Curcuma longa extract was studied at a wavelength range 230 to 550nm
and three groups are present in this plant like D-n-butyl disulfide, Anthracene and Acetyl chloride.

Table IX: UV-Spectrum peak value and its functional groups of Curcuma longa.

S. No Peak Area Functional groups

1. 235.90 Acetyl chloride, 2-Bromothiophene
2. 247.01 Di-n-butyl disulfide, Acetophenone 2,3 dimethyl
3. 370.95 Anthracene
4. 464.45 Thioester

Figure IX: UV Spectrum of Ethanol extracts of Curcuma longa.

CONCLUSION
The present study reveals the ethanol extract of mixture of four medicinal plants was an effective natural antibacterial agent.
The plant extract was tested against different pathogenic bacteria species such as Staphylococcus aureus, Bacillus subtilis, E.coli and
Klebsiella pneumonia. The ethanol extract produced significant zone of inhibitions and sensitive to the growth of Bacillus subtilis, and
E.coli. From the results of this study it has been concluded that the plant extracts may have great potential as a remedy for infectious
disease caused by these pathogenic bacteria species. FT-IR analysis revealed the presence of the secondary alcohol, phenol, amines,
alkanes and halogen. UV –VIS analysis of Justicia tranquebariensis, Tribullus terrestris and Curcuma longa confirm the presence of
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Aniline. Alkenes, alkynes, aromatics and nitrosobutane are found in Justicia tranquebariensis, Tribullus terrestris, and Aloe vera.
Thus in future these plant could be used to extract and find a novel wound healing drugs since they are efficient in inhibiting the
microbes.
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ABBREVIATIONS
UV-VIS - Ultraviolet and Visible Spectrophotometer
FT-IR – Fourier Transform Infrared Spectrometry.

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CONFLICT OF INTEREST STATEMENT

The authors declare that there is no conflict of interest.

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