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Journal of Applied Sciences Research, 6(2): 139-143, 2010

© 2010, INSInet Publication

Anti-inflammatory and Analgesic Potential of Leaf Extract of Allium Stracheyi


1
Shashi Ranjan, 1 Vikash S. Jadon, 1 Nitish Sharma, 1 Kamini Singh, 1 Varsha Parcha, 1Sanjay Gupta
and 2J.P. Bhatt

1
Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Balawala,
Dehradun, Uttarakhand, India, 248161
2
Department of Zoology and Biotechnology, HNB Garhwal University, Srinagar, Uttarakhand, India

Abstract: Allium stracheyi, a medicinal plant found at the height of 2500-3000 meters of Alpine
Himalayas of Uttarakhand, India is used as a spice and in ethano-medicine by local people. In the present
study, Allium stracheyi leaves were explored for their anti-inflammatory and analgesic potential on
experimental model and compared to standard drugs. The results showed that methanolic extract has
significant reduction in inflammation i.e. 61% (100mg/kg) (p< 0.5) as compared to the standard drug
Diclofenac sodium suspension in 0.1% Tween 80 (10 mg/kg body weight) as well as the highest analgesic
potential i.e. 64.62% (100mg/kg) as compared to aspirin 68.62 % at 25 mg/kg body weight. Thus,
methanolic extract of the plant can be fully explored for its anti-inflammatory and analgesic potential
which has not been reported so far. The plant extract showed a relative low toxicity hence justifies the
folkloric use of plant by the local people in W estern Himalayan region for curing inflammation and
painful conditions.

Key words: Allium stracheyi, anti-inflammatory activity, analgesic activity, Phytochemical screening

INTRODUCTION might serve as lead for the development of novel


drug [7 ] .
The importance of medicinal plants in traditional Allium stracheyi (Jambu) is a perennial herb,
healthcare practices, providing clues to new areas of flowers are white in color about 35-40 centimeters in
research and in biodiversity conservation is now well height and is traditionally being used by the local
recognized. The use of traditional medicine and people as spice for flavoring. It is mainly found at the
medicinal plants in most developing countries, as a height of 2500-3000 meters of Alpine Himalayas of
normative basis for the maintenance of good health has Uttarakhand, India near moist rock, dry rock and steep
been widely observed. However, information on the slope with a strong preference of sunny site. Edible
uses for plants for medicine is lacking from many plant part used includes flowers, leaves, root and bulb.
interior areas of Himalayan regions in India. Utilization The leaf and bulb parts of this plant are used locally in
of plants for medicinal purposes in India has been the alleviation of inflammation and painful conditions [8 ].
documented long back in ancient literature. However, Leaves and inflorescences are also used as seasoning
organized studies in this direction were initiated in agents. Although no specific mention of medicinal uses
1956 and off late such studies are gaining recognition has been for this species, member of this genus are in
and popularity due to loss of traditional knowledge and general very healthy additions to the diet. They contain
declining plant population. sulphur rich compounds with antioxidant, anti-
Drugs which are in use presently for the inflammatory, and antimicrobial properties. Thus, in the
management of pain and inflammatory conditions are present study attempts were made to investigate its
either narcotics or non-narcotics (NSAIDS) and present anti-inflammatory and analgesic potential with a view
well known side and toxic effects[1 ,2 ,3 ] . On the contrary of justifying the use of this plant in treatment of
herbal medicines with good absorption, less toxicity inflammatory disease and analgesic locally.
and easy availability have been used since long [4 ]. It is
therefore essential that efforts should be made to M ATERIALS AND M ETHODS
introduce new medicinal plants to develop cheaper and
effective drugs[5 ,6] . Plants represent still a large Collection of Plant M aterials: W hole plant of Allium
untapped source of structurally novel compounds that stracheyi was collected from various parts of

Corresponding Author: Shashi Ranjan, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research,
Balawala, Dehradun, Uttarakhand, India, 248161
E-mail: shashiranjan16@gmail.com, Mobile: +91-9997581583
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J. Appl. Sci. Res., 6(2): 139-143, 2010

Uttarakhand, India during August to September 2008. carrageenan-induced rat paw edema method [1 1 ]. Twenty
The seed samples were submitted for conservation at five albino rats of either sex were taken and divided
National Gene bank, National Bureau of Plant Genetic into 5 groups, each group contained 5 rats. Group I –
Resources, Pusa Campus, New Delhi, India and the received Tween 80 solution (0.1%) in a volume of 10
national identity number (SR-01-IC 567645) was ml/kg body weight, Group II- received Diclofenac
obtained. The leaves of this plant were dried under sodium suspension in 0.1% Tween 80 (10 mg/kg body
shade at 27 ° C - 30 ° C for 15 to 30 days, after which weight), Group III- received Petroleum ether extract
the leaves of the plant were chopped and grounded into suspension in 0.1 % Tween 80 (100mg/kg body
coarse powder. weight), Group IV - received aqueous extract
suspension in 0.1 % Tween 80 (100mg/kg body
Preparation of Plant Extract: The coarse powder of weight) Group V – received methanol extract
material (200gm) was successively extracted with suspension in 0.1% Tween 80 (100mg/kg body
solvents in order of increasing polarity like petroleum weight) in a volume of 10ml/kg, half hour after
ether, chloroform, methanol and water using hot soxhlet carrageenan (0.1 ml of 0.1 %) administration in the sub
extractor. The resulting extracts were concentrated by plantar region of left hind paw of each rat. They were
vacuum evaporator keeping the maximum temperature deprived of water during experiment to ensure uniform
48 ° C to 50 ° C. hydration and to minimize variability in oedematous
response (W inter et al. 1962). The measurement of the
Phytochemical Screening: The presence of various paw volume (cm 3 ) was done on the principle of
phytochemical constituents in the extract was vo lum e d isp lac e m e nt usin g L E T I C A d ig ital
determined using standard screening tests [9 ]. Plethysmometer. The readings were taken before and at
30 min intervals after the injection of carrageenan for
Experimental Animals: W istar rats (150-200g) and a period of 4 hrs. The edema at each time was
Swiss Albino M ice (25-40g) of either sex were used calculated in relation to the paw volume before the
for this study. The animals were obtained from the injection of the carrageenan.
animal house of the Department of Pharmacy, Sardar
Bhagwan Singh (P.G) Institute of Biomedical Sciences The anti- inflammatory activity was determined as
and Research, Dehradun, Uttarakhand, India. The the percentage of inhibition of inflammation after it
animals were maintained under standard environmental was induced by carrageenan by taking volume of
condition and had free access to food and water before inflammation in control group as 100%. The percentage
administration of plant extract inhibition was calculated by using the formula:

Acute Toxicity study (LD 5 0 ): The intraperitoneal (i.p) % Inhibition =Mean paw inflammation of control –
acute toxicity of the Methanolic extract was evaluated M ean paw inflammation of test / Mean paw
in Swiss Albino Mice by method described by inflammation of control x 100.
Lorke[1 0 ] . The method involved the determination of
LD 5 0 value in biphasic manner. The animals were Acetic Acid-induce W rithing in M ice: The analgesic
starved of feed but allowed access to water 24 hours investigation was carried out according to the
prior to the study. In the initial investigatory step method[12 ] . The mice were divided into six different
(phase 1), a range of doses of the extract producing the groups (of four mice each). They were differently pre-
toxic effects was established. This was done by treated with the different extract (100mg/kg i.p), aspirin
intraperitoneal administration of widely differing doses (100mg/kg i.p) and Control normal saline (10ml/kg i.p)
of the extract (50, 100, 500, 1000, 1200mg/kg i.p) to 30, 60, 90 and 120 min after the treatment, 1% acetic
four groups of mice (of four each). Based on the result acid solution was administered to the mice
obtained, a phase II investigatory step was done by (1ml/100gm, i.p). They were placed in a transparent
giving more specific doses (350, 400, 450, 500 mg/kg observation box. Five minutes after the administration
i.p) to four other groups of mice. of acetic acid the number of abdominal constrictions
The mice were observed for 24 hours for such (writhes) made within 20 minutes of every mouse was
behavioral signs as nervousness, excitement, dullness, counted. The results of the treatment groups were
ataxia or death. The LD 5 0 was estimated from the compared with those of normal saline pre-treated
geometric mean of the dose that caused 100% mortality control. The percentage of the writhes was calculated
and the dose which caused no lethality. as:

A nti-inflam m a to ry A ctivity: Anti-inflammatory % of writhes = (Test mean/Control mean) x 100.


activity of Allium stracheyi was evaluated by

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J. Appl. Sci. Res., 6(2): 139-143, 2010

Statistical Analysis: The calculation of the average revelation of the median lethal dose LD 5 0 of the
edema for the anti-inflammation and percentage Methanolic extract of A. stracheyi to 445.5 mg/kg i.p
writhing reflex for anti-nociception were based on the is probably an indicator that the extract may be
expression of numerical data as mean ± SEM. The relatively safe where as estimated LD 5 0 value > 1g is
statistical significance between groups were analyzed considered safe[1 0 ] . This is of importance because a
using two-way analysis of variance (ANOVA), cumulative toxic effect could not occur if the extract is
followed by students t-test. p values < 0.5 were taken taken over time. In the present study, the abdominal
to be significant. constriction (W rithing) model adopted is thought to
partly involve local peritoneal receptors [1 3 , 1 4 , 1 5 ]. The
Results: ability of the methanolic extract to cause a significant
Phytochemical Screening: The results of qualitative reduction in the number of acetic-acid induced writhes
analysis indicated that petroleum ether and chloroform in mice probably suggests an anti-nociceptive property.
extract is rich in steroids while methanolic and aqueous The use of abdominal constriction (writhing) model is
extract is rich in alkaloid and saponins (Table 1). known to be very sensitive when compared with other
models such as tail flick model[14 , 1 6 ] . This study also
Anti-inflammatory Activity: In the study the pro- showed that anti-nociceptive effect of the aqueous and
inflammatory effect was observed for methanol, chloroform extract showed very less effect in same
petroleum ether and aqueous decoction against the duration of time as compared to methanolic extract.
standard Diclofenac sodium. Out of the three extracts The result obtained complimented the earlier
Methanolic extract showed significant percent reduction investigation that methanolic extract of Allium species
(61%) (p<0.5) as compared to the diclofinac sodium contains copious amount of flavonoids, organosulphur
that showed 72% inhibition of paw volume (Figure 1). compounds[1 7 ] . Previous study on different Allium
Non-significant inhibition was observed for petroleum species also shows that these flavonoids and
ether extract and aqueous extract with 46% and 20% organosulphur compounds are potent analgesic and anti-
inhibition when the dose administered was 100mg/kg. inflammatory agents. These compounds prevent the
formation of pro-inflammatory messengers. This is in
Acetic Acid-induced W rithing: The result represented line with some other reports that some metabolites are
in Figure 2, shows that all the extracts of Allium as potent or even more potent in some activities as the
stracheyi at the doses of 100mg/kg and aspirin parent compound.
25mg/kg exhibited significant (p<0.5) inhibition of the The carragenan induced edema model showed the
control writhes at the rate of 68.62%, 55%, 64.62% anti-inflammatory property of the extract. Although the
and 38.47 for aspirin, petroleum ether extract, mechanisms of action for the anti-nociception and anti-
methanolic extract and aqueous extract respectively inflammation are not yet elucidated, the combination of
when compared to that of control. both effects can be taken advantage of, therapeutically.
In addition, extracts of Allium stracheyi at the Significantly (p<0.5) high anti-inflammatory activity of
above mentioned doses, potentiated analgesic activity methanolic extract (100 mg/kg body weight) of A.
of aspirin shown by further decreasing the writhing stracheyi may be due to inhibition of the mediators of
response when given in combination. Statistical analysis inflammation such as histamine, serotonin and
showed that Methanolic extract has highest analgesic prostaglandin. Thus, the research work justified the
potential than petroleum ether extract of 64.62% of traditional use of the plant as a spice in food
inhibition as compared to the standard aspirin that gave supplements and in the treatment of pro-inflammatory
68.62% inhibition. disease. Further studies will be carried out on
pharmacodynamics pattern to establish the mechanism
Acute Toxicity Study (LD 5 0 ): Mice treated with doses of the action of the plant extracts.
$ 500 mg/kg i.p. were dull, panted, showed occasional
abdominal stretching and died within 24h of treatment. ACKNOW LEDGM ENTS
The adverse signs and deaths were however, not seen
at doses below 500mg/kg i.p. The LD 50 of Methanolic The authors wish to acknowledge the financial
extract of Allium stracheyi was estimated to be 445.5 support of The Director, Uttarakhand State Council of
mg/kg i.p. Science and Technology, Dehradun, Uttarakhand, India.
The authors would also like to express their gratitude
D isc ussio n: T he prese n c e o f a lk a lo id s a nd to The Management, Sardar Bhagwan Singh PG
organosulphur compounds, which has been indicated in Institute of Biomedical Sciences & Research, Balawala,
analgesic and anti-inflammatory activities, supports the Dehradun, India, for providing research facilities to
traditional and local use of Allium stracheyi. The conduct the work.

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J. Appl. Sci. Res., 6(2): 139-143, 2010

Table 1: Phytochem ical analysis of different extracts of Allium stracheyi


Test perform ed Petroleum ether extract Chloroform extract M ethanol extract Aqueous extract
1. Test for steroids
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
i. Salkowski test (+) (+) (-) (-)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
ii. Gilberm an-Buchard's test (+) (+) (-) (-)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
2. Test for alkaloids
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
i. W agner's test (-) (-) (+) (+)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
ii. H ager's test (-) (-) (+) (+)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
3. Test for phenolic and flavonoid com pounds
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
i. Vanilin-H CL test (-) (-) (+) (+)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
ii. Ferric chloride test (+) (+) (+) (-)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
iii. Zinc hydrochloric acid reduction test (-) (-) (-) (-)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
4. Test for tannins (-) (-) (+) (+)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
5. Test for saponins (-) (-) (+) (+)

Fig. 1: Anti-inflammatory activity of different extract of Allium stracheyi in Carrageenan induced paw edema in
rat compared to the same point in control group (Normal saline)

Fig. 2: Analgesic effect of different extract of Allium stracheyi (100mg/kg, ip) & Aspirin (25mg/kg ip) in 1%
acetic acid induced paw licking in rat compared to the same point in control (Saline)

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