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DOI:
10.4103/jdras.jdras_85_22
Department of Pharmacy,
1
Rajiv Gandhi Proudyogiki
Vishwavidhyalay, Bhopal,
2
Department of Zoology,
Sri Satya Sai University of
Technology and Medical
Sciences, Sehore, Madhya
Pradesh, India
Address for
correspondence: Abu
Tahir,
Department of
Pharmacy, Rajiv
Gandhi Proudyogiki
Vishwavidhyalay,
Bhopal 462033,
Madhya Pradesh, India.
E-mail: aliabutahir2009@
gmail.com
Submitted : 07-06-2022
Revised : 04-09-2022
Accepted : 21-11-2022
Published : 31-03-2023
© 2023 Journal of Drug Research in Ayurvedic Sciences | Published by Wolters Kluwer - Medknow 173
Original Article
Evaluation of the anti-inflammatory
activity of hexane and ethanolic
extracts of polyherbal formulation
of Nigella sativa L. (seeds), Ocimum
tenuiflorum L. (leaves), and Piper
longum L. (fruits) on carrageenan-
induced paw edema in wistar rats
Abu Tahir1, Mohd Shafi Dar2
Abstract
BACKGROUND: Inflammation is a primary physiological defence mechanism that assists the
body in protecting itself from infection, toxic chemicals, or other noxious stimuli. The current study
demonstrated phytochemical screening, anti-inflammatory activities, and sub-acute toxicity of hexane
and ethanol extracts of Nigella sativa L. (seeds), Ocimum tenuiflorum L. (leaves), and Piper longum
L. (Fruits), as well as anti-oxidant activity. The Carrageenan-Induced Rat Paw Edema method was
used to assess anti-inflammatory activity.
METHODS: The carrageenan-induced paw edema test evaluated anti-inflammatory activity. Male
albino Wistar rats weighing 150 ± 10g were divided into six groups of six animals each. Paw edema was
induced with 1.5% carrageenan in all the groups except the normal. Group, I received a plain control of
1 ml of 1% Carboxy Methyl Cellulose (CMC); Group II standard drug received Indomethacin (10 mg/kg);
Group III received Polyherbal formulation of hexane extracts (PHFH) 250 mg/kg b.w; Group IV received
PHFH 500 mg/kg b.w.; Group V was given 250 mg/kg b.w. of Polyherbal formulation of ethanolic extracts
(PHFE). The extract’s acute toxicity (2000 mg/kg) as per OECD guidelines was studied in albino rats
for 14 days. The qualitative analysis of various phytochemical constituents of various phytoconstituents
was determined. The DPPH method was used to evaluate anti-oxidant activity.
RESULTS: The results showed that both PHFH and PHFE exhibited marked inhibition of the edema
size from 1, 3, and 5 hrs of study as compared to the standard drug indomethacin (10 mg/kg b.w).
The PHFE (250 and 500 mg/kg) displayed excellent protection against inflammation to PHFH (250
and 500 mg/kg). Compared to the standard drug, indomethacin which showed the highest excellent
protection against inflammation. PHFE has lower anti-oxidant activity than standard ascorbic acid but
exhibits higher anti-oxidant activity than PHFH. In an acute toxicity test, hexane-ethanolic extracts
up to 3000 mg/kg had no toxic effects.
CONCLUSION: From this study, we conclude that Nigella sativa L. (seeds), Ocimum tenuiflorum
L. (leaves), and Piper longum L. (fruits) have anti-inflammatory activity by reducing paw inflammation
as well as showing anti-oxidant activity.
Keywords:
Anti-inflammatory, antioxidant, iper longum, nigella sativa, ocimum tenuiflorum, polyherbal formulation
This is an open access journal, and articles are distributed under How to cite this article: Tahir A, Dar MS. Evaluation
the terms of the Creative Commons Attribution-NonCommercial- of the anti-inflammatory activity of hexane and
ShareAlike 4.0 License, which allows others to remix, tweak, and ethanolic extracts of polyherbal formulation of Nigella
build upon the work non-commercially, as long as appropriate credit sativa L.  (seeds), Ocimum tenuiflorum L.  (leaves),
is given and the new creations are licensed under the identical terms. and Piper longum L. (fruits) on carrageenan-induced
paw edema in wistar rats. J Drug Res Ayurvedic Sci
For reprints contact: WKHLRPMedknow_reprints@wolterskluwer.
2023;8:173-80.
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Tahir and Dar: Evaluation of the anti-inflammatory activity Nigella sativa L., Ocimum tenuiflorum L., and Piper longum L.
Introduction
T he term ‘inflammation’ comes from the Latin word
‘inflammo,’ which means ‘to set alight; to ignite, or
to set on fire.’ Inflammation is referred to as a hot thing in
Greek. Within, the Greek word phlegmon has been used
to define inflammatory lesions.[1,2] Inflammation has
always been a mystery to humans. This phenomenon
can be triggered by something as minor as a bruise or as
serious as a myocardial infarction. Non-steroidal anti-
inflammatory drugs (NSAIDS) and corticosteroids are
the most commonly used anti-inflammatory drugs, but
their toxic side effects include epigastric distress, peptic
ulceration, osteoporosis, and iatrogenic Cushing’s
syndrome limited their use.[3,4] It is a normal reaction to
disruptions in homeostasis caused by infection, injury,
or trauma.[2,5-7] Waram (inflammation) is a broad term
in Unani medicine that refers to any abnormal swelling
caused by the accumulation of blood, pus, water, or
flatus.[8] A swelling known as a waram is brought on
by an abnormal substances absorbing into an organ.[9,10]
According to the WHO, 70–80% of the world’s population
relies on non conventional medicine, primarily from
herbal sources, for primary health care.[11,12] Its popularity
is growing, particularly in developing countries where
the cost of consulting a physician and the cost of medicine
are out of reach for the majority of people.[13]These anti-
inflammatory medications are used to relieve pain in
various conditions such as arthritis, muscle pain, and
ligament pain. Conventional drug treatments are not
very exclusive in controlling the occurrence and outcome
of many inflammatory diseases. In addition, they cause
significant side effects in patients.[14]
Herbal medicines derived from plant extracts are
increasingly being used to treat a wide range of clinical
diseases, despite little being known about their mode
of action. The pharmacological evaluation of various
plants used in Indian traditional systems of medicine is
gaining popularity.It is known that Ayurvedic herbals
are prepared in many dosage forms, mostly of which
are PHF (Polyherbal formation).In the current scenario,
Plant-derived medicinal compounds such as flavonoids,
saponins, alkaloids, terpenoids, glycosides, and
coumarins could provide an excellent starting point for
developing new anti-inflammatory agents that are more
efficacious, safer, more affordable, and more accessible
to patients.
The plant has been linked to various pharmacological
activities, including diuretics, CNS depressants,
diaphoretic and styptic properties. Plant decoction has
activity in renal and urinary complications. Plant seeds
have anti-infammatory activity, and plant roots are used
to treat cancer and scrofulous tumours.[15,16]Carrageen-
174 Journal of Drug Research in Ayurvedic Sciences - Volume 8, Issue 2, April-June 2023
induced rat paw edema exhibits numerous biochemical
and cellular characteristics that have been clearly
identified in the background and are constantly being
revised in light of discoveries.[17,18]
Based on the nature of the interaction, there are
two mechanisms for how synergism acts (i.e.,
pharmacodynamics and pharmacokinetic). In terms of
pharmacokinetic synergism, the ability of the herb to
facilitate the absorption, distribution, metabolism and
elimination of other herbs is focused. On the other hand,
pharmacodynamic synergism studies the synergistic
effect when active constituents with similar therapeutic
activity are targeted to a similar receptor or physiological
system. Other than that, it is believed that multiple
factors and complications cause diseases in most cases,
leading to visible and invisible symptoms. Here, the
combination of herbals may act on multiple targets at
the same time to provide thorough relief.[19]
Due to synergism, polyherbalism confers some
benefits unavailable in a single herbal formulation.
Better therapeutic effect can be reached with a single
multi-constituent formulation. For this, a lower dose
of the herbal preparation would be needed to achieve
desirable pharmacological action, thus reducing the
risk of deleterious side effects. Besides, PHFs improve
patients’ convenience by eliminating the need to take
more than one different single herbal formulation at a
time, which indirectly leads to better compliance and
therapeutic effect. All these benefits have resulted in
the popularity of PHF in the market when compared
to single herbal formulation.[20]This study is aimed to
evaluate anti-oxidant and anti-inflammatory effect
of PHF prepared by the combination of Nigella sativa
(seeds), Ocimum sanctum L.  (leaves) and Piper longum
L. (fruits) hexane and ethanol extracts individually
in different doses on carrageenan-induced rat paw
edema model.This approach is an alternative method
to discover potentially new herbal combinations/
formulations for the treatment of inflammation and its
symptoms.
Material and Methods
Plant materials and authentication
Plant materials of Nigella sativa L.  (seeds), Ocimum
tenuiflorum L.  (leaves), and Piper longum L.  (fruits)
were collected locally from Bhopal, M.P. And the
authentication of all the three plants were done by Dr. Zia
Ul Hasan, Department of Botany, Safia College of Science,
Peergate, Bhopal, M.P India. A  voucher specimen of
Nigella sativa L. (seeds), Ocimum tenuiflorum L. (leaves),
and Piper longum L.  (fruits) were deposited for record
with the specimen number 332/Bot/Safia/18, 366/Bot/
Safia/18 and 418/Bot/Safia/18, respectively.
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Tahir and Dar: Evaluation of the anti-inflammatory activity Nigella sativa L., Ocimum tenuiflorum L., and Piper longum L.
Extraction and preparation of hexane and
ethanolic extracts of each selected plant
Material
The seeds of Nigella sativa L., leaves of Ocimum tenuiflorum
L., and fruits of Piper longum L.  were air-dried up to
7 days. Dried plant parts were stored in air-tight glass
containers in dry and cool place to avoid contamination
and deterioration.[21] Each air-dried medicinal plant
(100 gm) was coarsely powdered and extracted by cold
extraction method (double maceration) with hexane
and ethanol at 25°C for 48 hours. The extracts were
individually filtered through laboratory filter paper,
and the filtered solutions were dried using a rotary
evaporator under reduced pressure (335 mbar) at 40 ± 2°C
and stored in the refrigerator (2–4°C) for further activity.
Preparation of polyherbal formulations (suspension)
Dried hexane extract of Nigella sativa L. (seeds), Ocimum
tenuiflorum L. (leaves), and Piper longum L. (fruits) were
mixed in 1:1:1 ratio and suspended in double distilled
water with carboxymethyl cellulose sodium (CMCS)
(2%) to prepare polyherbal formulation of hexane
extracts (PHFH). Similarly, dried ethanolic extractof
Nigella sativa L. (seeds), Ocimum tenuiflorum L. (leaves),
and Piper longum L. (Fruits) were mixed in 1:1:1 ratio and
suspended in double distilled water with carboxymethyl
cellulose sodium (CMCS) (2%) to prepare polyherbal
formulation of ethanolic extracts (PHFE).[22]
Preliminary phytochemical analysis
The phytochemical anaylsis of hexane and ethanol
extracts of Nigella sativa L. (seeds), Ocimum tenuiflorum
L. (leaves), and Piper longum L. (fruits) was performed
in 100 ml of its own mother solvents to obtain a stock
with a concentration of 1% (v/v). The resulting extracts
were subjected to preliminary phytochemical screening
using standard phytochemical screening methods[23,24]
Experimental animals
Wistar rats (150 ± 10 g) were housed in groups of six (n = 6)
under a standard 12 h light/dark cycle and temperature and
humidity control (25 ± 2°C, 55–65%). Rats were fed standard
rodent chow and given unlimited access to water. Before the
experiments, rats were acclimatised to laboratory conditions
for 7  days. The experimental work was conducted at
Pinnacle Biomedical Research Institute (PBRI), Bhopal,
India. With protocol approval reference number was PBRI/
IAEC/PN-18010. The animal studies were approved by the
Institutional Animal Ethics Committee (IAEC), which was
established by the Ministry of Environment and Forests,
Government of India, for the purpose of controlling and
supervising experimental animals (CPCSEA Reg No.-1824/
PO/ERe/S/15/CPCSEA).
Acute oral toxicity
Acute toxicity testing of plant materials prepared
extracts was performed following the Organization
Journal of Drug Research in Ayurvedic Sciences - Volume 8, Issue 2, April-June 2023 175
 
for Economic Cooperation and Development (OECD)
Guidelines-423.[25] The animals were fasted for four hours
but had free access to water the entire time. According to
OECD recommendations, the starting dose level should
be that which is most likely to cause mortality in some of
the dosed animals; and when no information is available
on a substance to be tested in this regard; for animal
welfare reasons, the dose level to be used as the starting
dose is chosen from one of three fixed levels of 50, 100,
300, and 2000 mg/kg body weight. The selected animals
were alienated into 4 different groups. All animals were
kept as per conditions mentioned in the guidelines. The
dose selected for both (PHFH, PHFE) was 2000 mg/
Kg. The variations in consciousness, abnormalities,
rate of respiration, skin colour, behaviour, excretion,
impairment in diet intake, water feeding, and loss of hair
or death of test animals were observed first 5 h, 12 h and
every day for 14 days.[26]
In vitro anti-oxidant assay
DPPH assay method
2 ml solution of both PHFH and PHFE in different
concentrations (20  μg/ml-100  μg/ml) were added
individually to 2 ml of DPPH solution (75 μM). Standard
solution was ascorbic acid in the same concentrations as
samples. The reaction mixtures were shaken thoroughly
and kept in the dark for 30 minutes. The solution was
prepared by adding methanol (2 ml) to DPPH solution
(2 ml).[27] The absorbance of all the samples and control
solution was measured at 517nm using UV-Visible
spectrophotometer and the remaining DPPH was
calculated. The radical scavenging activity was expressed
as the percentage inhibition and was calculated using the
following formula:
Percentage of Inhibition = [(AC - AS)/AC] X 100.
Where AC is the absorbance of the control and AS is the
absorbance of the Sample. The graphs were plotted
between % inhibition and different concentrations of
PHFH, PHFE and standard ascorbic acid individually
and IC50 value was determined.
Chemicals and drugs
Indomethacin (purchased from a local pharmacy store),
Carrageenan (Sigma Aldrich, India), All of the chemicals
used in this experiment were of the analytical grade.
Design of Experiment
Six groups of six animals were formed at random from
the animals. Group I control group receives Carrageenan
1% (sub-plantar injection) + 0.5 ml of Carboxymethyl
cellulose sodium (CMCS) (2%), orally.Group II Standard
group receives Carrageenan 1% (sub-plantar injection) +
Indomethacin (10 mg/kg b.w), orally. Group III and IV
Treatment groups receives Carrageenan 1% (sub-plantar
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Tahir and Dar: Evaluation of the anti-inflammatory activity Nigella sativa L., Ocimum tenuiflorum L., and Piper longum L.

injection) + PHF prepare by hexane extract (250 mg/ Table  1: Anti-inflammatory activity of polyherbal
kg and 500 mg/kg), orally. Group V and VI Treatment formulations (PHFH and PHFE)
groups receives Carrageenan 1% (sub-plantar injection) S. No Treatment Paw volume (ml) (mean ± SEM)
+ PHF prepare by ethanolic extract (250 mg/kg and 1h 3h 5h
500 mg/kg), orally respectively. 1 Control 1.662 ± 0.122 1.922 ± 0.045 2.42 ± 0.087
group
Anti-inflammatory activity (in-vivo) 2 Standard 0.795 ± 0.091 0.69 ± 0.094 0.4525 ± 0.078
group
Carrageenan-induced paw edema (CIPE)model 3 Treatment 1.412 ± 0.062 1.917 ± 0.066 1.800 ± 0.082
Evaluation of anti-inflammatory activity of both PHFH group PHFH
and PHFE was performed on male Wistar albino rats 250mg/kg
(150 ± 10g) by CIPE model. Animals of all groups 4 Treatment 1.480 ± 0.025 1.617 ± 0.128 1.552 ± 0.0425
were administered with 0.1 ml sub-plantar injection group PHFH
500mg/kg
of carrageenan (1%) in normal saline in the left hind
5 Treatment 1.262 ± 0.092 1.487 ± 0.120 1.375 ± 0.072
paw to induce acute inflammation, 1 h after oral drug group PHFE
treatment. [28] The volume of paw of each rat was 250mg/kg
measured by digital plethysmometer at 1,3 and 5 h after 6 Treatment 1.195 ± 0.072 1.285 ± 0.0967 1.06 ± 0.0353
administration of carrageenan injection.[29]The variation group PHFE
between initial and the successive readings represented 500mg/kg
the rate of inflammation or edema volume and the All values are stated as SD, number of animals in each group=6.
percent inhibition were measured.[22]
Table  2: Qualitative phytochemical analysis of
The following equation was used to calculate the percent ethanolic extracts
edema inhibition: Phytoconstituents Ocimum Nigella Piper
tenuiflorum L. sativa L. longum L.
 Vt  Steroids + + +
Percentedema inhibition=  1-  ×100 Tannins + + -
 Vc  Terpenoids + - +
Flavonoids + + +
Where, Vc represented the mean increase in paw volume Saponins - + +
of control group, and Vt represented the mean increase Alkaloids - + +
in paw volume of test and standard drugs. The results Glycosides + - +
were presented as SD Phenolics + + +
Fatty acids - - -
Statistical analysis Carbohydrate - + +
All values were expressed as mean ± SD (n = 6 in each Proteins - + +
group). One-way ANOVA was applied to test for the + represents presence of constituents; -represents absence of constituents
significance of biochemical data of the different groups.
P-values less than 0.05 were regarded as statistically Table  3: Qualitative phytochemical analysis of hexane
significant. extracts
Phytoconstituents Ocimum Nigella Piper
Results tenuiflorum L. sativa L. longumL.
Steroids + + +
Results of preliminary screening of phytochemicals and Tannins + - -
in vitro anti-oxidant assay by DPPH method along with Terpenoids + + +
anti-inflammatory activity by carrageenan-induced paw Flavonoids - + -
edema method were analyzed statistically and presented Saponins - - -
in Tables 1-5 and Figures 1-5. Alkaloids - - -
Glycosides + - -
Discussion Phenolics + + +
Fatty acids + + +
The carrageenan-induced rat paw edema model is an Carbohydrate - - -
appropriate test for evaluating anti-inflammatory drugs, Proteins - + +
and it has frequently been used to assess the drug’s anti- + represents presence of constituents; -represents absence of constituents
edematous effect. Carrageenan is a powerful chemical
that stimulates the release of inflammatory and pro- Acute or chronic inflammation is the two types of
inflammatory mediators (prostaglandins, leukotrienes, inflammation. Acute inflammation is the body’s initial
histamine, bradykinin, TNFα, and so on).[30,31] response to injurious stimuli, manifested by increased
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Tahir and Dar: Evaluation of the anti-inflammatory activity Nigella sativa L., Ocimum tenuiflorum L., and Piper longum L. 

Table  4: DPPH scavenging activities of extracts of

PHFH, PHFE andAscorbic acid
concentrations μg ml-1 % inhib ition
% inhibition
PHFH PHFE Ascorbic
acid
70
(Standard) 60
20 μg ml-1 35.14412 44.0133 52.21729 50
40 μg ml-1 38.3592 47.22838 55.87583
40
60 μg ml-1 42.1286 50.99778 61.08647
30
80 μg ml-1 51.21951 58.42572 68.62528
100 μg ml-1 60.53215 66.85144 71.3969
20 y = 0.3182x + 26.386
IC50 = IC50 = IC50 = 13.725 10 R² = 0.9456
74.27μg 47.74 μg μg ml-1 0
ml-1 ml-1
0 50 100 150
Values were expressed as mean ± SD for triplicates
Concentration (µg/ml)
Table  5: Percentage inhibition in anti-inflammatory
activity of polyherbal formulations (PHFH and PHFE) Figure 2: DPPH assay by using PHFH
S. No Treatment Percent
Inhibition (%)
1
2
Control group
Standard group
--
81.40
% inhibition
3 Treatment group PHFH 250 mg/kg 25.6 80
4 Treatment group PHFH 500mg/kg 36
5 Treatment group PHFE 250mg/kg 43.24 60
6 Treatment group PHFE 500mg/kg 56.20
40

20 y = 0.2844x + 36.441
% inhibition 0
R² = 0.9567

0 20 40 60 80 100 120
80
Concentration (µg/ml)
60
Figure 3: DPPH assay by using PHFE

40
y = 0.2555x + 46.508 2.5
20 R² = 0.9816
2

1 hr
0 1.5
3 hr
0 50 100 150 1
5 hr
Concentration (µg/ml) 0.5

Figure 1: DPPH assay by using standard ascorbic acid
movement of plasma and leukocytes from the blood into
the injured tissues. Acute inflammation is initiated by
cells already present in the tissues. This is characterized
by significant vascular changes, such as vasodilatation
and increased capillary permeability, caused by the
actions of various inflammatory mediators.Chronic
inflammation is a long-term inflammatory response that
results in a progressive shift in the type of cells present
at the site of inflammation and is characterized by the
inflammatory process’s simultaneous destruction and
healing of tissues.
0
Control Standard PHFH 250 PHFH 500 PHFE 250 PHFE 500
Figure 4: Paw volume at different hours 1, 3 and 5 of PHFH and PHFE at 250 mg/
kg and 500 mg/kg
The response to carrageenan-induced paw edema is
biphasic.[32-34]
The first phase was mediated by the release of
histamine, serotonin, and kinin, whereas the second
phase was mediated by the release of prostaglandins
and slow-acting substances, which peaked at 4 hours.
According to reports, the second phase is sensitive to
both steroidal and non-steroidal anti-inflammatory

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Tahir and Dar: Evaluation of the anti-inflammatory activity Nigella sativa L., Ocimum tenuiflorum L., and Piper longum L.
90
80
70
60
50
40
30
20
10
0 The results showed that both PHFH and PHFE were
Control Standard PHFH 250 PHFH 500 PHFE 250 PHFE 500
Figure 5: Percentage inhibition of PHFH and PHFE at 250 mg/kg and 500 mg/kg
agents.[35] Prostaglandins are the primary cause of acute
inflammation. NSAIDs, in general, strongly inhibit the
second phase of carrageenan-induced edema, whereas
others inhibit both phases.The ability of a compound to
reduce local edema induced in the rat paw by injection
of an irritant agent is the most widely used primary test
for screening new anti-inflammatory agents. N.  sativa
(seeds), O. sanctum L. (leaves) and P.longum (fruits) may
contain anti-inflammatory agents that are responsible for
the inhibition of prostaglandins and the inflammatory
pathway. Anti-inflammatory activity of polyherbal
formulations was evaluated by carrageenan-induced rat
paw edema method and outcomes are given in Table 1.
The anti-inflammatory activity of Nigella sativa L. (seeds),
Ocimum tenuiflorum L.  (leaves), and Piper longum
L. (fruits) hexane and ethanol extracts may be due to a
combination of different biological constituents rather
than any single compound.The presence of secondary
metabolites such as alkaloids, flavonoids, glycosides,
phenols, saponins, sterols, and others may contribute to
the curative properties of medicinal plants. Tables 2 and 3
show the presence of flavonoids, glycosides, phenols,
fatty acids, steroids, and terpenoids in successive extracts
of Nigella sativa L. (seeds), Ocimum tenuiflorum L. (leaves),
and Piper longum L. (fruits).
Under acute toxicity test, oral administration of both
the polyherbal formulations (PHFH, PHFE) at the dose
of 2000 mg/ Kg body weight exhibited no evidence of
any abnormalities, significant behavioral changes, skin
sensitivity or deaths. This may be due to the fact that
all the three plants Nigella sativa L.  (seeds), Ocimum
tenuiflorum L. (leaves), and Piper longum L. (Fruits) have
been traditionally used for many years by humans to
treat various disease ailments without any reported
incidence of adverse effects.
Figures 1–3 showed the anti-oxidant activity of the
polyherbalformulations compared with ascorbic acid. IC50
values of ascorbic acid, PHFH and PHFE were 13.725μg/
ml, 74.27μg/ml and 47.74μg/ml, respectively.According
to the findings, PHFE has lower anti-oxidant activity
178 Journal of Drug Research in Ayurvedic Sciences - Volume 8, Issue 2, April-June 2023
than ascorbic acid but higher anti-oxidant potential than
PHFH [Table 4]. The scavenging effect of polyherbal
formulations(PHFH, PHFE) on DPPH might be due to
the hydrogen contributing abilityof active constituents
present in plant extracts.[36] Though, the scavenging
potential of both the PHFs were found less than that of
ascorbic acid,the results supported the proton-donating
potential of both the PHFs as primary anti-oxidants.
exhibited appreciable inhibition of the edema size from
1, 3, and 5 hrs of study as compared to the standard
drug indomethacin [Figure 4]. The PHFE emerged as
the most promising formulation in doses of 250 and
500mg/kg [Figure 5], also displayed excellent protection
against inflammation with percent protection of 43.24%
and 56.20%, respectively [Table 5]. The PHFH also
showed considerable inhibition of the edema size in both
doses of 250 and 500mg/kg [Figure 5], also exhibited
protection against inflammation with percent protection
of 25.6%and36.0%, respectively [Table  5]. However, the
standard indomethacin (10 mg/kg) showed the highest
inhibition of edema size and showed remarkable protection
against inflammation with percent protection of 81.40%
protection [Table 5].
On the basis of above outcomes, it can be inferred that
the PHFE inhibitory effect against inflammation induced
by carrageenan is more potent when compared to
PHFH. It is postulated that the inflammatory potential
of both the polyherbal formulations (PHFE and PHFH)
may be due to the COX enzyme inhibition potential of
constituents present in plant extracts.[37] The study’s
findings revealed that various phytochemicals are
present in plant extracts and may be responsible for
pharmacological activity. Plant extracts reduces pain
in rats, leading to the conclusion that it has potent anti-
infammatory activity.
Conclusion
The antiinflammatory properties of Nigella sativa L. (seeds),
Ocimum tenuiflorum L. (leaves), and Piper longum L. (fruits)
were observed in present study. It is also proposed that
the mechanisms of action of Nigella sativa L.  (seeds),
Ocimum tenuiflorum L. (leaves), and Piper longum L. (fruits)
are related to the inhibition of histamine, serotonin,
and prostaglandin synthesis. However, more research
is needed to isolate and characterise anti-inflammatory
chemical constituents found in hexane and ethanolic
extracts of Nigella sativa L.  (seeds), Ocimum tenuiflorum
L. (leaves), and Piper longum L. (fruits).
Acknowledgement
The authors would like to express their gratitude to Dr.
Megha Jha,Sr. Scientific Manager (RandD)of PBRI, for
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Tahir and Dar: Evaluation of the anti-inflammatory activity Nigella sativa L., Ocimum tenuiflorum L., and Piper longum L.
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Tahir and Dar: Evaluation of the anti-inflammatory activity Nigella sativa L., Ocimum tenuiflorum L., and Piper longum L.

हिदं ी सारांश

विस्टर चूहों में कै रेगीनन प्रेरित पाव-एडिमा पर निजेला सैटिवा एल. (बीज), ओसिमम टेनइु फ्लोरम एल. (पत्र), और
पाइपर लोंगम एल. (फल) के बहुपादपीय योगो के हेक्सेन और एथेनॉलिक अर्क की शोथरोधी कारक का मूल्यांकन

पृष्ठभूमि: शोथ एक प्राथमिक शारीरिक रक्षा तं त्र है जो शरीर को सं क्रमण, जहरीले रसायनों या अन्य हानिकारक उत्तेजनाओं से बचाने में सहायता करता है। वर्तमान अध्ययन
में निगेला सैटिवा एल. (बीज), ओसिमम टेन्यूफ्लोरम एल. (पत्र), और पाइपर लोंगम एल. (फल) के हेक्सेन और इथेनॉल के अर्क की फाइटोके मिकल स्क्रीनिगं और
एं टी-ऑक्सीडेंट एक्टिविटी, एं टी इन्फ़्लेमेटरी एक्टिविटीज़ तथा सब-एक्यूट टोक्सिसिटी का प्रदर्शन किया गया। एं टी इन्फ़्लेमेटरी एक्टिविटी का आकलन करने के लिए
कै रे गीनन -प्रेरित चूहा पाव-एडिमा विधि का उपयोग किया गया ।
पद्धतियाँ: कै रेगीनन -प्रेरित पाव-एडिमा परीक्षण ने एं टी इन्फ़्लेमेटरी एक्टिविटी का मूल्यांकन किया। 150 ± 10 ग्राम वजन वाले नर अल्बिनो विस्टर चूहों को छह जानवरों
के छह समूहों में विभाजित किया गया। सामान्य को छोड़कर सभी समूहों में 1.5% कै रेगीनन के साथ पाव-एडिमा को प्रेरित किया गया। समूह I को 1% कार्बोक्सी मिथाइल
सेलुलोज (सीएमसी) के 1 मिलीलीटर का नियं त्रण प्राप्त हुआ; समूह II मानक दवा इं डोमेथसे िन (10 मिलीग्राम / किग्रा) प्राप्त हुई; समूह III ने हेक्सेन अर्क (पीएचएफ़एच)
250 mg/kg bw का पॉलीहर्बल सूत्रीकरण प्राप्त किया; समूह IV ने पीएचएफ़एच 500 mg/kg bw प्राप्त किया; समूह V को इथेनॉलिक अर्क (पीएचएफई) के
बहुपादपीय योगो को 250 mg/kg b.w दिया गया । अर्क की तीव्र विषाक्तता (2000 मिलीग्राम/किग्रा) प्रति ओईसीडी दिशानिर्देशों के अनुसार एल्बिनो चूहों में 14 दिनों
तक अध्ययन किया गया। विभिन्न पादप घटकों के विभिन्न पादप रासायनिक घटकों का गुणात्मक विश्लेषण निर्धारित किया गया। एं टी-ऑक्सीडेंट गतिविधि का मूल्यांकन
करने के लिए डीपीपीएच पद्धति का उपयोग किया गया।
परिणाम: परिणामों से पता चला है कि पीएचएफ़एच और पीएचएफ़ई दोनों ने मानक दवा इं डोमेथसे िन (10 mg/kg bw) की तुलना में 1, 3, और 5 घं टे के अध्ययन से
एडिमा के आकार का उत्कृ ष्ट अवरोध प्रदर्शित किया। पीएचएफ़एच (250 और 500 mg/kg) ने पीएचएफ़ई (250 और 500 mg/kg) को सूजन से उत्कृ ष्ट सुरक्षा प्रदान
की। मानक दवा की तुलना में, इं डोमेथसे िन ने सूजन के खिलाफ उच्चतम उत्कृ ष्ट सुरक्षा दिखायी। पीएचएफ़ई में मानक एस्कॉर्बिक एसिड की तुलना में कम एं टी-ऑक्सीडेंट
गतिविधि है, लेकिन पीएचएफ़एच की तुलना में उच्च एं टी-ऑक्सीडेंट गतिविधि प्रदर्शित करता है। एक एक्यूट टोक्सिसिटी परीक्षण में, 3000 मिलीग्राम/किलोग्राम तक
हेक्सेन-एथेनॉलिक अर्क का कोई विषाक्त प्रभाव नहीं था।
निष्कर्ष: इस अध्ययन से, हम निष्कर्ष निकालते हैं कि निगेला सैटिवा एल. (बीज), ओसिमम टेन्यूफ्लोरम एल. (पत्र), और पाइपर लोंगम एल. (फल) में एं टी इन्फ़्लेमेटरी
एक्टिविटी होती है, जो पं जे की सूजन को कम करने के साथ-साथ एं टी-ऑक्सीडेंट गतिविधि दिखाती है।
शब्दकुं जी: एं टी इन्फ़्लेमेटरी, एं टी-ऑक्सीडेंट, पाइपर लोंगम, निगेला सैटिवा, ओसिमम टेन्यूफ्लोरम, पॉलीहर्बल फॉर्मूलेशन

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