Int. J. Life Sci. Pharma Res.

2019 Oct; 9(4): (P) 90-102 ISSN 2250-0480

Original Research Article P lant physiology and biochemistry

International Journal of Life science and Pharma Research

COMPARING ANTIBACTERIAL POTENTIAL AND PHYTOCHEMICAL

CONSTITUENTSOF TWO SPECIES OF GENUS URTICA

PRIYANKA RAJPUT*, MAITRY CHOUDHARY AND R.A. SHARMA

Plant Physiology and Biochemistry Lab, Lab No. 14,Department of Botany, University of Rajasthan,
Jaipur, Rajasthan- 302004

ABSTRACT

The present study was conducted to evaluate and compare the in-vitro bioactive potential of various
organic extracts of the root, stem and leaves of Urtica dioica and Urtica urens on growth of tested Gram
Positive (+ve) and Gram negative (–ve) bacteria.The present investigation also includes the comparative
chemical constituency of the crude extract of root, stem and leaves of both plants. The comparative
bioactive potential of Urtica dioica and Urtica urens was tested against both pathogenic and non-
pathogenic strains of bacteria that are Gram positive (Staphylococcus aureus and Bacillus subtilis) and
Gram negative (Pseudomonas aeruginosa and E. Coli) conducted by using agar well diffusion method
.The Minimum Inhibitory Concentrations (MICs)-the lowest concentration of antimicrobial agent that will
inhibit any visible growth of microbein chloroform and methanol extracts of root, stem and leaves of both
plants was determined using micro-well dilution method. For the phytochemical evaluation of plants
metabolites- ascorbic acid, flavonoids, phenols and proteins has been quantitatively estimated. The MIC
has been to be most effective in case of methanolic extract of root against both gram positive and gram
negative bacteria. Significant antimicrobial activity has been observed against gram negative bacteria E.
coli and both the gram positive bacteria testedexpressed by U. dioica comparatively more effective than
U. urens. Only a moderate amount of activity is exhibited against P. aeruginosa by both the plants. The
non-polar extracts were found to have comparatively higher bactericidal potential than the polar extracts.
The phytochemicals analysis suggested the presence of ascorbic acid, phenols and proteins to be found in
U. urens in significantly higher concentration than U. dioica. On the other hand, U. dioica has been found
to be more abundant in flavonoid concentration comparatively.

KEYWORDS: Antibacterial efficacy, Urtica dioica, Urtica Urens, Agar well diffusion assay,
Minimum Inhibitory Concentration (MIC), Phytochemicals constituency.

PRIYANKA RAJPUT *
Plant Physiology and Biochemistry Lab, Lab No. 14,Department of
Botany, University of Rajasthan,Jaipur, Rajasthan- 302004

Received on: 03-10-2019

Revised and Accepted on: 31.10.2019
DOI: http://dx.doi.org/10.22376/ijpbs/lpr.2019.9.4.P90-102

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INTRODUCTION found helpful in reducing difficulties associated

with urination problems in benign prostatic
hyperplasia and also in regulating sugar levels in
A vast source of medicinally active plants has
diabetic patients.12Phytochemicals are the
been provided for the benefit of human beings by
metabolites of non-nutrient nature derived from
the environment.1 Thousands of modern drugs are
plants found in various dietary substances, food
isolated from these vast natural sources from the
grains, vegetables with an enormous potential of
knowledge and information of medicinal uses of
fighting the chronic human and animal disorders.
herbs that are scripted in our ancient literature of
These phytochemicals arise in the plant body
Ayurveda, Charak samhita. 2 Researchers are
through various in-built metabolic processes.13
continuously exploring the potentials of different
The phytochemicals analysis of genus Urtica has
plants and their products for expanding the use of
proved the presence of 100 different compounds
natural sources for isolating active biological
like sterols, phenols, fatty acids, lignans,
compounds making them suitable for
3 alkaloids, terpenoids and other compounds.
pharmacological usage. The genus Urtica is
Among all the species of the genus Urtica, the
derived from the word ‘euro’ which means to burn
most important one regarding to its importance in
or ‘urere’ to sting. 4Urtica dioica (wild
ethno-pharmacological medicines is U.
inhabitant), and Urtica urens (wild as well as 14
dioica. The current investigation has been
cultivated in recent years) belong to family
conducted to evaluate the comparative potential of
Urticaceae of order Rosales is commonly known
the two different species of the genus Urtica to act
as stinging nettle and dwarf or burning nettle,
against the bacterial growth and both nonpolar
respectively. The names have been provided
and polar extracts of U. dioica and U. urens were
probably on the basis of characteristic property of
evaluated to compute the MIC exhibited by
being covered with stinging hairs that introduce
different plant parts- root, stem and leaves. Apart
irritants onto the human skin on coming in contact
from screening the comparative bioactive
with the plant resulting in persistent pain and
potential of the plant, a comparative study on
burning sensation afterwards. The potential of this
phytochemical constituency of crude extract of
plant for long remained far from
both plants has been carried out.
acknowledgement and has remained undervalued
till now5. But now-a-days the plants, especially U.
urens has started gaining significance due to its MATERIALS AND METHODS
pharmacological potential. 6 U. dioica plant is a
perennial shrub, erect in nature, 1-1.5 m tall in Chemical Reagent
height armed with stinging hairs. Leaves are Petroleum ether, chloroform, acetone,
found in opposite arrangement and are 7-15 cm methanol,distilled water, sodium chloride, nutrient
long. Stinging hairs of U. dioica are trichomes agar, ciprofloxacin as standard, metaphosphoric
that are usually found distributed along its leaves acid, 2,4-dinitrophenylhydrazine, thiourea, copper
and stem, that acts as hypodermic needles which sulphate, sulfuric acid, ascorbic acid, aluminium
on coming in contact with skin injects chemicals chloride, potassium acetate, quercetin, Folin–
like acetic acid, histamine, etc. that causes a Ciocalteau reagent, caffeic acid, ethanol, Sodium
stinging sensation.6-8Urtica urens is an annual Carbonate (Na2CO3), Tri-chloro Acetic acid
herb with erect plant of ascending type up to 2 (TCA), alkaline solution.
feet tall about 60 cm in height with a tap root
system. Leaves on the plant found are oval with Collection of plant material and extraction of
opposite arrangement up to ½ to 2 inch or 4 cms plant sample for antimicrobial potential
long, sharp, tipped leaves that are densely toothed. estimation
The irritant hairs present on the leaves and stem of Urtica dioica and Urtica urens fresh plant
plant cause irritant dermatitis.9Traditionally the material were collected from the Dist.- Palampur
plant stinging potential has been used in therapies (Himachal Pradesh) (July, 2016) and Village –
known as urtication, which is believed to treat the Naggar, Dist-Kullu (H.P.) (May, 2017)
numb arthritic and paralytic limbs by flailing with respectively, and were authenticated by CSIR,
a fresh plant.10 This stimulates blood circulation in IHBT Herbarium, Palampur(H.P.). Voucher
the area and brings warmth to joints. Stinging specimen herbarium (PLP-18301) of Urtica
nettle is currently being used in the treatment of dioica and (PLP-18302) of Urtica urens were
rheumatism as a diuretic agent. 11 It is utilized in deposited at IHBT, CSIR, Palampur (H.P.)
relieving symptoms of seasonal allergies and also herbarium. The plant was thoroughly washed with
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distilled water without pressing or crushing it to method.15 Following the provided protocol the
remove dirt and soil particles. Root, Stem and medium used for bacterial growth was Mueller
leaves of plant were separated, shade dried and Hinton Agar Medium. The agar media was melted
then powdered. The dried powder (200gm) was and cooled to 48-50ºC before pouring into plates.
then extracted using different solvents- Petroleum Plates were prepared by pouring 25 ml of freshly
ether, chloroform, acetone, methanol and water by prepared agar media in sterilized 100mm X 15
soxhlet extraction apparatus. The solvent extract mm plates. Plates were allowed to solidify.The
was prepared by dissolving 20 mg of powdered solidified agar plates were then inoculated
material sequentially in 100 ml solvent each. The aseptically with various bacterial strains
solvent was allowed to get evaporated at room suspensions. The terget bacterial strain
temperature spontaneouslyand the remaining dry suspensions were freshly prepared by diluting the
crude extract from each of the solvent was microbial culture to prepare a microbial
weighed and then diluted in 100% DMSO at a concentration of 108 CFU/ml. Wells of 3mm
concentration of 10 mg/ml. radius were prepared in the inoculated agar
plates.The analyte or the material under testing
Bacterial Strains (60 µl each well) was then introduced in the wells
The strains of bacteria against which the effective (6 mm) prepared. These plates were then kept
potential of various solvents of plant parts was under incubation at 37ºC for 24 hours. The
tested includes Staphylococcus aureus (MTCC antibacterial potential were measured in terms of
0087), Pseudomonas aeruginosa (MTCC 4646), the diameter of the inhibition zone produced by
Bacillus subtilis (MTCC 0121), E. coli (MTCC the chemical composition of the analyte tested for
1652). These bacterial strains are collected from the potential activity in comparison to that of
S.M.S. Medical College, Jaipur, Rajasthan, India. antibiotic, ciprofloxacin of concentration 10
These strains were maintained in sterile nutrient mg/ml taken as standard (40 µl each well). As the
agar (Himedia) slants and were stored at 4ºC until incubation period is over, the plates were analyzed
further use . for the inhibitory zones (ZI) measured in
millimeters (mm). The extracts of U. dioica and
Agar well diffusion assay U. urens exhibiting significant inhibitory effect on
In-vitro antibacterial efficacy of different plant microbial growth were shown in figure 1 below.
parts were tested using agar well diffusion

Zone of Inhibition of Sample

Activity Index (AI) =
Zone of Inhibition of Standard
(1)
The activity index of the extract would be determined using the above described formulae.

MIC determination micro plate, 5 μl inoculum and 95 μl of nutrient

For the determination of MIC, micro-plate method broth was added, to which 100μl extracts of
was used.16The MICs of the chloroform and concentrations 100 μg/ml, 50 μg/ml, 25 μg/ml,
methanol extract of the root, stem and leaves of U. 12.5 μg/ml, 6.25 μg/ml and 100 μl of TTC (0.01%
dioica and U. urens were determined. In this w/v). The plates were incubated at 37 ºC for 24
method TTC (2, 3, 5-triphenyltetrazolium hrs in incubator. In the presence of bacterial
chloride) visually indicates the presence of growth the TTC converts to red colored formazan
microbial growth. In each well of the 96-well which indicates the active viability of cells.

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Figure 1
Illustration of zone of Inhibition (selected) exhibited by various organic extracts

a.S. aureus: UD:S(P) g. B. subtilis: UD: S (C) m. E. coli: UD: L(C)

b.S. aureus: UD: S(C) h. B. subtilis: UU: S(P) n. E. coli: UU: S(P)
c. S. aureus: UD: L(C) i. B. subtilis: UU: S (C) o. E. coli: UU: R(A)
d.S. aureus: UU: S (P) j. B. subtilis: UU: R(P) p. P. aeruginosa:UD:S(C)
e. S. aureus: U.U: S (M) k. E. coli: UD: S(P) q.P. aeruginosa:UD:S(M)
f. B. subtilis: UD: S(P) l. E. coli: UD: S(C) r.P. aeruginosa:UU:R(M)
Key: UD →U. dioica, UU→U. urens S→ Stem, L→ Leaves, R→ Root, P→ Pet Ether, C→
Chloroform, M→ Methanol, A→ Acetone.

Phytochemical Analysis metaphosphoric acid in 6.0 gm/dl concentration.

The quantitative estimation of phytochemicals: After 5 minute homogenized material was
ascorbic acid content, flavonoids, phenols and centrifuged at 2500 rpm for 10 minutes.
protein contentin different plant parts of U. dioica Supernatant was collected in separate test tube of
and U. urens was carried out The detailed which 1.2 ml was taken and 0.4 ml of 2,4-
procedure were described below. dinitrophenylhydrazine-Thiourea-Copper sulphate
reagent was added into it. The mixture was
Total ascorbic acid content incubated for 2 hours at 37º C and then chilled for
The total ascorbic acid content of powdered plant 10 min in an ice bath. To each test tube 6 ml of
samples were estimated using a slight cold sulfuric acid (12mol/l) was mixed. Now the
modification of the colorimetric method.17 Each solution was divided into three separate test tubes
of the powdered plant material (500 mg) was of 3.6 ml each and the optical density of the
homogenized in 20 ml of freshly prepared mixture was measured at 520 nm through

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spectrophotometer. 1.2 ml of metaphosphoric acid 1M Potassium acetate solution and 5.6 ml of

6.0 gm/dl concentration was stated as blank. The distilled water. Then the optical density of each
concentration of the test samples were computed prepared extract was measured at 415 nm. The
from the standard calibration curve Figure 2 (a) of solution prepared by substituting sample with 0.25
the ascorbic acid. ml ethanol dissolved in 3 ml methanol, 200 μl of
10% AlCl3 solution, 200 μl of 1M Potassium
Total Flavonoid Content acetate solution and 5.6 ml of distilled water serve
Total flavonoids content of an extract was as blank. The concentration of the test samples
determined using aluminium chloride colorimetric were computed from the standard calibration
method.18 According to this method 200 μg/ml curve of quercetin (Figure 2 (b)).The total
concentration of extract solution was prepared by flavonoid concentration is measured in terms of
adding 1 ml of methanol plant extract in 3 ml mg/100g QE (Quercetin Equivalent).
methanol, 200 μl of 10% AlCl3 solution, 200 μl of

R × D. F.× V × 100
Total Flavonoid Content =
W

R = Concentration computed through standard curve of quercetin

V = Volume of stock Solution
D.F.= Dilution factor
100=for 100 gm dried plant
W= Weight of the plant used in experiment (in gm)

Estimation of total protein content concentrations of protein in different extracts

The total protein of the powdered extract has been provided in figure 2 (c).
estimated by using Lowry method.19 According to
this method, 10 gm of plant sample was weighed Total Phenolic Content Determination
properly and homogenized in 50 ml of 10% TCA. Phenolic content present in the samples was
After that obtained extract was centrifuged for 10 determined by using Folin–Ciocalteu reagent
min at 15000 rpm in 4ºC, of which the supernatant using caffeic acid as a standard phenolic
was discarded and further pellet was dissolved in compound.20 According to this method 200 mg of
5% TCA. Mixture was mixed in a vortex mixer plant sample crushed with 3ml of 80% ethanol
and collected in a test tube. The collected mixture was centrifuged for 20 minutes at 1500 rpm at
was kept in incubator at 80 ºC for 30 minutes. The room temperature. From the supernatant 1 ml is
mixture is allowed to cool at room temperature. taken in the test tube to which 1 ml of one
From the sample extract mixture 5 ml is separated milliliter of Folin–Ciocalteu reagent was added
and to it is added 25 ml of alkaline solution and and the content of the flask was mixed thoroughly.
2.5 ml of folin-ciocalteu phenol reagent and it is After 3min, 3ml of Na2CO3 was added which then
mixed properly. The optical density of sample was allowed to stand for 2h with intermittent
mixture was taken at 750 nm via a UV-Vis shaking. The absorbance was measured at 750
spectrophotometer with 10% TCA taken as blank nm in a spectrophotometer. The total
solution. A standard calibration curve of Bovine concentration of phenolic compounds in the
Serum Albumin (BSA) of varying concentration ethanol extract was determined as micrograms of
with their respective optical density readings at caffeic acid equivalent by using an equation that
750 nm has been used for calculating the was obtained from standard caffeic acid graph
given in figure 2 (d).

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Figure 2
Calibration curves of (a) Ascorbic acid (b) Quercetin (c) BSA (d)Caffeic acid

STATISTICAL ANALYSIS RESULTS

Antibacterial screening
Statistical analysis of data was performed using The antibacterial assay of different extracts of
minitab SPSS statistical subscription. One way root, stem and leaves of U. dioica and U. urens on
ANOVA (Analysis of variance) was carried out observation after 24 hr incubation at 38 ºC
for statistical analysis. Each experiment was exhibited various zones of inhibition with their
repeated 3 times. Mean±Standard error was respective activity index compiled in comparative
computed from analysis of each treatment. Data graphs (Figure 3). From the above data obtained
was presented in mean±SE format and were via agar well diffusion method, some of the
compared via Tukey’s test at a level of 5% extracts show high inhibitory activity against
probability. growth of various gram positive as well as gram
negative bacteria.

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Figure 3
Graphs indicating comparative activity indices of U. dioica and U. urens exhibited by various organic
extracts of (a) Root, (b) Stem and (c) Leaves

The significantly higher amount of inhibitory U. dioica, spirit and water extract of stem and
activity has been exhibited by petroleum ether leaves chloroform extract of U. dioica exhibits
extract of root and leaf of U. dioica and stem of significant inhibitory activity against S. aureus
U. urens, chloroform extracts of root and leaves of (gram positive). The plant is found to be least
U. dioica, acetone extract of all the tested plant effective against P. aeruginosa, except the
parts of U. dioica and root of U. urens while all chloroform extract of stem and leaves of U. dioica
the leaf extracts of U. urens shows moderate plant shows moderate inhibitory activity against
effect against gram negative bacteria E. coli. the growth of P. aeruginosa. Pet ether extract of
Significant inhibition effect was also observed root, chloroform extract of stem and leaves,
from pet ether extract of root, stem and leaves of methanol extract of root, stem and leaves of U.

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dioica shows least significant inhibitory potential MIC Determination

against gram positive bacteria B. subtilis. On the The methanol and chloroform found to be the
other hand, chloroform extract of root, pet ether most effective against most of the tested bacteria.
and chloroform extract of stem and aqueous Therefore both the extracts were examined for the
extract of the leaves of both U. dioica and U. minimum concentration required for antimicrobial
urens shows moderate inhibitory potential against growth. The result of MIC determined that
both B. subtilis and P. aeruginosa. Apart from all methanol extract of root was active against all
the above observations an overall comparatively four strains of microorganisms at the lowest
more effective inhibition on growth of all the concentration examined i.e. 6.25 mg/ml as shown
tested microorganisms were impacted by U. in table 1. The extracts that shows significant MIC
dioica rather from U. urens.The methanol extract values were chloroform extract of root against B.
of the root of U. dioica showed significant level of subtilis, methanol extract of stem and leaves
inhibitory potential against all gram positive and against E. coli. The minimum MIC value was
negative bacteria taken for testing. B. subtilis has exhibited by root chloroform extract against P.
been found to be sensitive to both methanol and aeruginosa and against B. subtilis by methanol
chloroform extract of stem at a concentration of extract of stem and leaves. All the remaining
100 mg/ml. No significant difference has been extracts showed an intermediate effect against
found in the inhibitory potential of the methanol remaining bacterial strains.
and chloroform extract of stem and leaves against
P. aeruginosa.
Table 1
MIC determination of chloroform and methanol samples of roots leaves and stem at
concentrations ranging from (6.25 to 100 mg/ml)
Name of the Minimum Inhibitory Concentration
Bacteria Root Stem Leaves
U. dioica U. urens U. dioica U. urens U. dioica U. urens
M C M C M C M C M C M C
E. coli 6.25 12.5 NA 50 6.25 12.5 NA 100 6.25 50 NA NA
P. 6.25 100 NA 25 12.5 12.5 25 NA 12.5 12.5 NA 12.5
aeruginosa
B. subtilis 6.25 6.25 12.5 12.5 100 100 NA NA 50 12.5 NA NA
S. aureus 6.25 12.5 NA NA- 12.5 25 NA 25 12.5 50 12.5 12.5
#‘NA’ → No activity .
Phytochemical analysis
Primary phytochemical investigation was done to determine the chemical composition of the different
plant part crude extract. The comparative results of the appropriate quantitative estimation tests
conducted were compiled in graphical format below in figure 4.

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Figure 4
Comparative graphs of quantitative estimation of different plant parts of U. dioica and U. urens
(a)Ascorbic acid (b) Flavonoids (c) Protein (d) Phenols

Ascorbic acid estimation Protein Estimation

Ascorbic acid content found in crude powdered The total protein concentration of the sample
extracts in roots, stem and leaves of U. dioica and material was computed from standard calibration
U. urens computed from the standard ascorbic curve of (Bovine Serum Albumin) BSA prepared
acid calibration curve formed by observing optical in different concentrations at 750 nm wavelength
density of standard ascorbic acid at varying in a UV-Vis spectrophotometer. Comparatively
concentration at 520 nm wavelengths via a UV- significant amount of protein concentration in
Visible spectrophotometer. The highest amount of their chemical constitution was shown by all plant
ascorbic acid concentration 26.439±0.891 μg/ml parts of U. urens of which leaves shows the
was exhibited by root extract of U. urens.All the highest amount of 280.4±35.2 μg/ml whereas
plant parts has higher proportion of ascorbic acid least amount 8.488±0.671 μg/ml was found to be
in their chemical composition.The results of total available in the leaves of U. dioica. The obtained
ascorbic acid concentration in different plant part results were compiled in table 2.
extracts of U. dioica and U. urens were tabulated
in table 2.
Table 2
Total concentration of ascorbic acid and protein ( in μg/ml)

Plants Root Stem Leaves

a b
U. dioica Ascorbic acid Concentration 10.42±0.648 14.615±0.582 8.488±0.671a
(μg/ml)
Protein Concentration 85.39±4.37b 98.58±7.53a 115.7±6.81c
(μg/ml)
U. urens Ascorbic acid Concentration 26.439±0.891b 18.919±0.642c 23.555±0.628b
(μg/ml)
Protein Concentration 175.4±32.4a 185.5±38.42a 280.4±35.2c
(μg/ml)
# Values are mean± S.E. (standard error) of 3 observations (p<0.05). Values marked with similar
superscript a, b or c indicates lack of significant difference from each other

Total Phenolic and Total Flavonoid Content extract i.e. 168.393±5.781 mg CAE/100gdw.
Both the total phenolic and flavonoid The range of concentration of flavonoid and
concentration of different parts of plant are shown phenols found in U. dioica is 22.522±3.515to
in table 3. The highest concentration of flavonoid 76.007±2.483 mg QE/100gdw and
has been reported from leaf extract i.e. 126.75±4.9321 to 168.393±5.781 mg
76.007±2.483 mg QE/100gdw while highest CAE/100gdw, respectively.
phenol concentration was reported from root

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Table 3
Total flavonoid and total phenol concentration

Plant Total flavonoid content in mg QE/100g of Total phenolic content in mg CAE/100g of

Part dried material dried material
U. dioica U. urens U. dioica U. urens
Root 22.52±6.09a 20.77±2.84a 168.39±10.01a 154±9.86b
Stem 68.211±3.380b 36.214±0.972a 126.75±10.46b 260.97±17.02c
b c a
Leaf 76.007±2.483 5.280±2.86 153.683±3.424 260.7±25.3c
# Values are mean± S.E. (standard error) of 3 observations (p<0.05). Values marked with similar
superscript a, b or c indicates lack of significant difference from each other

DISCUSSION the treatment of B. cereus, V. parahaemolyticus,

S. aureus and methicillin-resistant S. aureus
The results of the conducted study exhibited a (MRSA) infections. As it is well known, MRSA,
greater more significant microbial growth E. coli and Bacillus species, especially B. cereus,
inhibitory activity by organic extracts of U. are the potential food poisoning agents. The
dioica. The bactericidal activity was more active potential area of applying plant extracts in the
against gram negative bacteria E. coli and reduction in numbers and growth inhibition of
moderate activity against gram positive bacteria food-borne pathogens.26 For introducing these
B. subtilis and S. aureus. The results suggested extracts for future use in pharmaceutical and food
comparative higher resistance of gram –ve industries further research and testing were
bacteria P. aeruginosa against almost all the needed to examine the effectiveness as well as the
tested organic extracts of both plants U. dioica toxicity of these extracts to achieve the pure
and U. urens.This difference in antibacterial bioactive component that attribute to their
property towards P. aeruginosa was attributed to significant antimicrobial potential. Antimicrobial
the morphology of bacteria which is the presence activity of plant extract was attributed to the
of an extra protective membrane of presence of numerous bioactive compounds,
lipopolysaccharide enclosing the cell wall in gram therefore the phytochemical screening of plant
–ve bacteria that help in blocking the penetration parts has been done along with quantitative
of bioactive compounds.21-23It has been observed determination of total phenols and flavonoid
that growth of gram positive bacteria was most concentration in the sample.Terpenes and phenols
sensitive due to the presence of phenols as a of U. dioica are found to be one of the major
predominant active compound in the plants of groups of compounds with a potential source for
genus Urtica.24The various organic extracts of extraction of useful drugs exhibiting the inhibitory
different plant parts of both plants exhibited great activity of microbial infections.27-30In accordance
spectrum of inhibitory activities by agar well to the findings of the present study, research
diffusion method. The results indicated that the shows that antimicrobial activity was found in U.
non-polar extracts like petroleum ether extracts dioica water extract against E. coli, while no
has been significantly more active against significant activity was found against P.
microbial growth. Results of some studies shows aeruginosa. On the other hand, in contrast to our
that non-polar extracts provide better results than predicted results a research showed that U. dioica
polar extract.25According to the present research exhibits low antibacterial potential against S.
results petroleum ether and chloroform extracts aureus, whereas no activity against E. coli, P.
showed higher antimicrobial activity than the aeruginosa and S. aureus 31 and another study
other crude organic extracts. A comparatively confirmed that U. urens possess no significant
larger zone of inhibition was observed against antibacterial activity.32-34The significant effect of
E.coli (chloroform extract of root of U. dioica), inhibition on E. coli growth can be hypothesized
with an MIC value of 12.5mg/mL while the as the effect that an extract exerted on the outer
methanol extract of the root has an MIC value of membrane leads to alteration in membrane
6.25 mg/ml. The plant U. dioica has been used in structure and permeability of the cell. These
changes may result in breakage of hydrogen
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bonds that help in keeping the structure of toxicity testing of the extracts showing significant
membrane rigid.35,36 It is a fact that gram positive bactericidal activity should be done before
bacteria are comparatively more susceptible to introducing them for usage to commercialize in
inhibition by the solvent extracts than their gram – the form of pharmaceutical medicine.
ve counterparts.In addition, our findings
suggested that the non-polar extracts like ACKNOWLEDGMENTS
petroleum ether and chloroform of U. dioica and
some polar and non-polar extracts of U. urens that The authors are thankful to Head, Department of
are showing good inhibitory effect on growth of Botany, University of Rajasthan, Jaipur, India for
providing laboratory facilities and the Senior
pathogenic bacteria may further be suggested as a
principal scientist, IHBT, CSIR Palampur for the
potential source of natural antimicrobial agent.
identification of the plant specimen.

CONCLUSION
AUTHORS CONTRIBUTION
The results obtained from the agar well diffusion STATEMENT
assay has established that the root, stem and
leaves of U. dioica plant display in-vitro Miss. Priyanka Rajput conceptualized and
antimicrobial potential to varying extent against gathered all the results and data by performing
various bacterial strains. The antimicrobial all the required experiments by her related to
potential was found better against gram positive
this work. Dr. R. A. Sharma provided
bacteria than towards gram negative ones. The
positive results of the anti-bacterial screening necessary inputs and guidance for the
contribute for the extraction of potent concerned work. All the authors discussed the
antimicrobial agents. It provides preliminary methodology, results and contributed the final
information for further phytochemical and manuscript.
pharmacological analysis on the chemical
constituency of the plant extracts. Further research CONFLICT OF INTEREST
is needed for the bioactive compounds
identification and isolation and also an in vivo Conflict of interest declared none.
antimicrobial activity evaluation along with

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