University Journal of Creativity and Innovative Research 2020 Vol-01 Issue-01

Phytochemical Studies and Antibacterial Activities of Ruellia tuberosa L.

Khin Nwe Than Khin Myint Maw
Yadanabon University Mandalay University of Distance Education
drkhinnwethannn@gmail.com drkhinmyintmawbot@gmail.com

Abstract antidote activity. These plants are used to cure antidote

for snake-bite, centipede and gecko-bite according to [2].
The medicinal plant Ruellia tuberosa L. of The preliminary phytochemical investigation was
Acanthaceae family is a tropical plant widely carried out by ethanol extracts for study of plants
distributed in South East Asia. In phytochemical containing chemical constituents (alkaloid, steroid,
investigation, 10 compounds were studied. Among them, flavonoid, glycoside, tannin, reducing sugar, phenolic
glycoside, alkaloid, reducing sugar, terpene, phenolic compound, saponin, polyphenol and terpenoid).
compound, polyphenol, saponin and tannin were Antibacterial activities were also made with various
present but flavonoid and steroid were absent in the extract (ethanol, ethyl- acetate and n-hexane) of the
powder of study plant. Moreover, the antibacterial whole plant of R. tuberosa L. by using agar well
activity were carried out by using 95% ethanol, ethyl- diffusion method against six different types of test
acetate and n-hexane. Ethanol extract, n-hexane extract organisms. The present research aimed to investigate the
and ethyl-acetate extract were obtained the best chemical composition of Ruellia tuberosa L. as well as
inhibition zone diameter (0.7 cm) on Salmonella typhi. to determine the antibacterial activity.
The inhibition zone of ethanol extract showed the most
effective on Staphylococcus aureus (0.8 cm) and less 2. Materials and Methods
inhibition zone showed on Pseudomonas aeruginosa
(0.3 cm). All of the bacteria strains except in E. coli 2.1. Morphological Studies
were showed sensitive to different plant extracts. E. coli
strain was resistant to all of the plant extracts (n-hexane, The specimens of Ruellia tuberosa L. were collected
ethyl-acetate and ethanol). These are not showed from Amarapura Township in Mandalay Region. The
inhibition zone on plant extracts. specimens were collected during flowering and fruiting
periods. The collected specimens were identified by the
Keywords: Ruellia tuberosa L., Phytochemical, help of available literature [4], [6] and [8]. The
Antibacterial activity morphological characters were made from the fresh
specimens of both the vegetative and reproductive parts.
1. Introduction
2.2. Phytochemical Studies
Ruellia tuberosa L. belonging to the family
Acanthaceae is growing abundantly in shade places The plant samples were extracted with ethanol
along the roadside. It is well known plant as Byauk and (EtOH) and distilled water. Before phytochemical
as well as Na-ga-hmaing in traditional medicine of investigation, the powdered of Ruellia tuberosa L. were
Myanmar society. In other country, its also known as stored in air tight container to prevent moisture changes
minnie root, fever root, snapdragon root and sheep and contamination. Ethanol and distilled water extracts
potato plant is a species of flowering plant in of Ruellia tuberosa L. were tested by using reagents to
Acanthaceae family. These plant names are also called the solution. After the tests were detected by visual
popping pod, duppy gun and cracker plant come from observation of color change or by the precipitate
the fact that children like to play with the dry pods that formation according to [7]. General reactions in these
pop when rubbed with spit or water [10]. analyses revealed the presence or absence of these
The Ruellia species are used medicinally to cure compounds in the crude extracts tested at Department of
gonorrhea, syphilis, eye sores and in renal infections. In Chemistry, University of Mandalay.
traditional medicine, Ruellia tuberosa L. are used to
cure diuretic, anti-diabetic, antipyretic, antihypertensive 2.3. Antibacterial Studies
and antidote [1]. The whole parts of genus Ruellia
tuberosa L. are used in bladder stones and in bronchitis. 2.3.1. Preparation of Crude Extracts
Paste of the leaves are also used for skin diseases and
boil. Roots are used as anthelmintic. In Bangladesh, The plant materials were washed with water, air
Ruellia tuberosa is used for treatment of gonorrhea [9]. dried at room temperature and then grinded to get fine
In Myanmar, the leaves of Ruellia tuberosa L. are powder and stored in air-tight containers. The whole
used to cure ulcers, boil, blain. The liquid of the drained plants of Ruellia tuberosa L. were extracted by using
leaves used to cure for impetigo in children skin disease. hot continuous extraction technique in a soxhlet
The whole plant parts of the Ruellia tuberosa L. have extraction method by using 95% ethanol and water.
Then, the extracts were evaporated by placing them on
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hot water bath at 50°C. All obtained extracts were on both surfaces; petioles 0.3-1.1 cm long.
weight before packed in water proof plastic flacks and Inflorescences axillary or terminal dichasial cymes.
stored at 4°C until. Flowers bisexual, zygomorphic, pentamerous, pale blue
or violet, 3.5-4.0 cm in diameter at anthesis, bracts
2.3.2. Determination of Antibacterial Activity linear. Calyx 5 partite, linear-lanceolate, persistent in
Microorganisms fruit. Corolla 5-lobed, infundibuliform, subequal.
Stamens 4, didynamous, included, attached to the base
The test organism (Bacillus cereus, Escherichia coli, of corolla tube; filaments filiform, white; anthers
Staphylococcus aureus, Shigella boydii, Salmonella dithecous, basifixed. Ovary oblongoid, glabrous,
typhi, Pseudomonus aeruginosa) used in this study. All bilocular with many ovules in each locule on the axile
the test strains were obtained from Microbiology placentae; style filiform; stigma bifid. Fruits capsule,
Laboratory, Biotechnology Department, Mandalay 1.0-2.9 cm long, oblongoid, mucronate at the apex,
Technological University. Before testing the bacteria green, brown colour in dry. Seeds many, brown colour,
were grown in Nutrient broth at 37°C and maintained on ovoid, compress.
nutrient agar slants at 4°C.

Culture Media

Muller-Hinton agar for antibacterial susceptibility

were used. All culture media will be prepared and treated
according to the manufacture guide lines (Hi-Media) [3].

Agar well diffusion method

The antimicrobial activity of Ruellia tuberosa

extract was studies against Staphylococus aureus,
Shigella boydii, Salmonella typhi, Escherichia coli,
Bacillus cereus and Pseudomonas sp.. Bacteria Figure 1. Habit of Ruellia tuberosa L.
pathogens were inoculated in Nutrient broth and
incubated for 6-8 hours at 37°C. These pathogen
cultures were swabbed throughout on nutrient medium
with the help of sterile cotton wool. Swab in three
directions to ensure complete plate coverage. Sterilize a
cork bore by autoclaving. Then, the agar well (0.9 cm in
diameter) were prepared by scooping out the media with
sterile borer. The extract solution (n-hexane, ethyl-
acetate and ethanol) was filled into each well (30 mg per
well) incubated at 37°C for 24 hours and one well filled
with solvent for control.

Antibacterial tested

The plates inoculated with different bacterial strains Figure 2. A branch with inflorescences and seeds of
were incubated at 37°C for 24 hours. Antibacterial Ruellia tuberosa L.
activities were evaluated by measuring inhibition zone
of complete growth inhibition surrounding bacterial 3.2. Phytochemical Studies
growth.
3.2.1. Phytochemical investigation of powder of the
3. Results whole plant Ruellia tuberosa L.

3.1. Morphological Studies Preliminary phytochemical investigation on the whole

plant of Ruellia tuberosa L. was done on the ethanol and
Family : Acanthaceae distilled water extracts and the presence or absence of
Scientific name : Ruellia tuberosa L. active constituents in this plant was shown in Table 1.
Local name : Byauk; Na-ga-hmaing
Common name : Minnine root, fever root
Perennial erect herbs with tuberous roots, up to 20-
35 cm high; stems and branches quadrangular, tumid at
the node. Leaves obovate, 1.9-8.6 m in length and 0.7-
3.6 cm in breadth, cuneate at the base, entire along the
margins, slightly acute at the apex, sparsely pubescent
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Table 2. Antibacterial activity of different solvent

Table 1. Preliminary phytochemical examination extracts from the whole plant of Ruellia tuberosa L.
of the whole plant of Ruellia tuberosa L. Control
Organisms A1 A2 A3
Result (Ethanol)
Solvent
Test Reagent Obser- Re-
Extract
vation mark Bacillus cereus 0.2 cm 0.5 cm 0.4 cm 0.6 cm
Wagner's Reddish
+
reagent brown ppt Escherichia
Alkaloid 1% HCl - - - -
Dragendroff's coli
Orange ppt +
reagent
EtOH, Mg Staphylococcus
95% Green 0.2 cm 0.4 cm 0.5 cm 0.8 cm
Flavonoid ribbon conc: – aureus
EtOH colour
HCl
1% FeCl3 and Shigella
95% Greenish 0.2 cm 0.6 cm 0.6 cm 0.5 cm
Polyphenol 1% K3 + boydii
EtOH blue colour
[Fe(CN)6]
Salmonella
95% Dil H2SO4,
Yellowish 0.3 cm 0.7 cm 0.7 cm 0.7 cm
Tannin brown + typhi
EtOH 1% FeCl3 Pseudomonas
colour ppt 0.3 cm 0.5 cm 0.7 cm 0.3 cm
Purplish aeruginosa
Phenolic Distilled 10% FeCl3
blue colour +
Compound Water Solution
- = no inhibition
Distilled water, A1 = Ruellia tuberosa L. with n-hexane extract
Distilled Yellow
Glycoside 10% lead + A2 = Ruellia tuberosa L. with ethyl acetate extract
Water colour ppt
acetate A3 = Ruellia tuberosa L. with ethanol extract
Reducing Distilled Benedict's Orange red
+
sugar water solution ppt
Distilled
Saponin NaHCO3 Frothing +
Water
CHCl3, acetic
Pep- Yellow
Terpene anhydride, +
ether ppt
conc: H2SO4
CHCl3, acetic
95% Yellow
Steroid anhydride, –
EtOH colour
conc: H2SO4
+ = Presence – = Absence ppt = precipitate

3.3. Antibacterial Studies

3.3.1. Antibacterial activity of ethanol, ethyl-acetate

and n-hexane extract of the whole plant of
Ruellia tuberosa L.
A. Bacillus cereus
The study of antibacterial activity was carried out by
using 95% ethanol, ethyl, acetate, n-hexane. The results
were shown in Table 2. According to this result, ethanol
extract, n-hexane extract and ethyl-acetate extract were
obtained the best inhibition zone diameter (0.7 cm) on
Salmonella typhi. The inhibition zone of ethanol extract
showed the best on Staphylococcus aureus (0.8 cm) and
less inhibition zone on Pseudomonas aeruginosa (0.3
cm). All of the bacteria strain except in E. coli was
showed sensitive to different plant extract. E. coli strain
was resistant to all of the plant extract (n-hexane, ethyl-
acetate and ethanol). These are not showed inhibition
zone.

B. Escherichia coli

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4. Discussion and Conclusion

In the present research work, the whole plants
Ruellia tuberosa L. family Acanthaceae with the
various extracts was carried out for phytochemical and
activity against some of the bacterial strains, This plant
was erect herbs with tuberous roots; leaves simple,
obovate; inflorescence dichasial cymes; flowers
bisexual, zygomorphic, pentamerous, pale blue or
violet; fruits capsule, oblongoid, mucronate at the apex.
These characters were in accordance with those
described by [4], [6] and [8].
The presence of the secondary metabolites in the
C. Staphylococcus aureus
crude extracts of this plant may be responsible for some
of the biological activity. The preliminary
phytochemical investigation on the whole plants of
Ruellia tuberosa L. contains alkaloid, terpene,
polyphenol, glycoside, phenolic compound, reducing
sugar, tannin and sponin but flavonoid and steroid were
absent.
According to antibacterial activity investigation,
ethanol extract more effective than those of the other
extract. The ethanol extract most effective on the
Staphylococcus aureus bacterial strain (0.8 cm). The
ethanol extract, n-hexane extract and ethyl-acetate
extract effective on Salmonella typhi (0.7 cm). E.coli
strain not effective on all plant extract.
D. Shigella boydii Escherichia coli to urinary tract infections,
neuonatal menigitis, septicemia, diarrhoea and
dysentery, Staphylococcus aureus to skin infections and
food poisoning, Shigella boydii to dysentery,
Salmonella typhi to cause systemic infections, typhoid
fever, Pseudomonas aeruginosa to pneumonia, septic
shock, urinary tract infection, gastrointestinal infection,
skin and soft tissue infections, Bacillus cereus to eye
infection, soft tissue and cutaneous infections,
according to [5].
In conclusion, Ruellia tuberosa L. contains potential
antibacterial and phytochemical components that may
be used for against various pathogens. To do this, the
three extracts were tested to the 6 bacterial species. This
plant crude extracts could serve as potential sources of
E. Salmonella typhi new antibacterial agents. However, further
investigations are required to identify the active
constituents and to verify the medicinal properties of
the active constituents.

Acknowledgements

I am greatly indebted to Dr Aung Aung Min, Rector

and Dr Si Si Khin and Dr Tint Moe Thuzar, Pro-rector
of Yadanabon University for their permission to submit
this research paper. I would like to express my thanks to
Dr Htar Lwin, Professor and Head and Dr Pyone Yi,
Professor, Department of Botany, Yadanabon
F. Pseudomonas aeruginosa University, for their permission to carry this research
work.
Figure 3. Antibacterial activity of the whole plant of
Ruellia tuberosa L.

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References
[1] A. R. Ahmad, A. Munin, B. Elya, "Study of antioxidant
activity with reduction of dppt radical and xanthine oxidase
inhibitor of the extract of Ruellia tuberosa L. leaf",
International Res. J. Pharm, 2012.
[2] Ashin Nagathein, "Pon Pya Say Ah Bea Don", Vol. 2,
(Myanmar version) Mingalar Press, Yangon, 1972.
[3] Atlas, R. M., Handbook of Microbiological Media,
London, 2004.
[4] Backer, C. A., Flora of Java, Vol. 2. Nordhoff. Graningen,
Nethrlands, 1965.
[5] Cruickshank, R.J.P., "Medicinal Microbiology",
Livingstone Ltd, London, 1975.
[6] Dassanayake, M.D., A Revised Handbook to the Flora of
Ceylon, Vol. 12. Amerind Publishing. Co. Pvt Ltd., New
Delhi, 1998.
[7] Harborne JB., Phytochemical methods, A guide to the
modern techniques of plant analysis. London: Chapman
and Hall, 1984.
[8] Hooker. J.D., The Flora of British India”, Vol. 4. Oxford-
Clarendon Press, 1885.
[9] K. Afzal, "Genus Ruellia: Pharmacological and
phytochemical importance in ethnopharmacology", Acta
Poloniae Pharmaceutica- Drug Research, Malaysia, 2015,
Vol. 72. No. 5, pp. 821 – 827.
[10] S. Kumr, P. Praveen and P. Pardhasaradhi, "Preliminary
phytochemical investigation and anti-ulcer activity of
aerial parts of Ruellia tuberosa L. (Acanthaceae) in male
Wistar rats;", Int J. Pharm Biomed Res, 2013, 4 (3), pp.
145-148.

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