A PROJECT REPORT ON ANALGESIC ACTIVITY ON AQUEOUS FRUIT

EXTRACT OF Tribulus terristrisSubmitted in Partial fulfillment of the requirement for the award of the degree of

Bachelor of Pharmacy By

C. SUDHAKAR REDDY

Department of Pharmacology

VASAVI INSTITUTE OF PHARMACEUTICAL SCIENCES,VASAVI NAGAR, PEDDAPALLI (V), SIDHOUT (M), NEAR BHAKARAPET RAILWAY STATION,

KADAPA 516247.

Analgesia(pain)

Pain has been classified as productive pain and non- productive pain while this distinction has no

physiologic meaning. It may serve as a guide to treatment productive pain has been described as a

warring of injury.

Analgesic work at the level of the nerves, either by blocking the signal from the peripheral nervous

system. Or by distorting the interpretation by the central nervous system . class of drugs , based on

type of pain, severity of pain, and risk of adverse effects traditionally, pain has been divided into two

classes,

1) Acute pain

2) Chronic pain Acute pain

Acute pain is self limiting in duration, and includes post operative pain, pain of injury, and

childbirth. Because pain of these types is expected to be short term and long term side effects of

analgesic therapy may routinely be ignored.

An important consideration of pain management of analgesic . This applies to both the narcotic

and non- narcotic analgesics in severe pain is that patients should not be subject to the return of

pain .

Chronic pain:

Chronic pain, pain lasting over three months and severe enough to impair

function, is more difficult to treat, since the anticipated side effects of the analgesics are

more difficult to manage. While some classes of drugs ,such as the narcotic

agonist/antagonist drugs bupronophine, nalbuphine and pentazocine, and the COX-2

inhibitors celecoxib and rofecoxib represent advance in reduction of adverse effects, they

are still not fully suitable for long term management of severe pain.

Classification of Analgesics

Analgesics

Mild Analgesics

Aspirin

Acetaminophen

Ibuprofen

Strong Analgesics

Opium

Codeine

Heroin

Symptoms

The early symptoms of CKD are the same as for many other illnesses. These

symptoms may be only sign of a problem in the early stages.Symptoms may include:

Appetite loss

General ill feeling and fatigue

Headaches

Itching (pruritus) and dry skin

Nausea

Weight loss without trying to lose weight

Symptoms that may occur when kidney function has gotten worse include:

Abnormally dark or light skin.

Bone pain.

Drowsiness or problems concentrating or thinking.

Muscle twitching or cramps.

Breath odour.

Complications

Acute renal failure.

Chronic renal failure.

Interstitial nephritis.

Renal papillary necrosis(tissue death)

Urinary tract infections, chronic or recurrent.

Hypertension.

Causes

Damage Analgesic involves nephropathy involves damage within the internal structures of

the kidney. It is caused by long term use of analgesic s, especially over the counter (otc)

medications that contain phenacetin acetaminophen and non-steroidal anti inflammatory

drugs(NSAIDS) such as aspirin or Ibuprofen .

Risk factors include :

Use of OTC analgesics containing more than one active ingredient .

Chronic headaches, pain full menstrual periods, backache, or musculoskeletal pain .

Emotional or behavioural changes .

History of dependent behaviours including smoking , alcoholism, and excessive use of

tranquilizers.

Diagnosis

Diagnosis is traditionally based on the clinical findings above in combination with excessive

analgesic use. It is estimated that between 2 and 3 kg each of phenacetin or aspirin must be

consumed before evidence of analgesic nephropathy becomes clinically apparent.

Once suspected, analgesic nephropathy can be confirmed with relative accuracy

using computed tomography (CT) imaging without contrast. One trial demonstrated that the

appearance of papillary calcifications on CT imaging was 92%sensitive and 100% specific

for the diagnosis of analgesic nephropathy.

Clinical features

Common findings in people with analgesic nephropathy include headache, anaemia, high

blood pressure (hypertension), and white blood cells in the urine (pyuria).

Clinical findings in analgesic nephropathy

Finding Proportion affected

Headache 35-100%

Pyuria 50-100%

Anemia 60-90%

Hypertension 15-70%

Gastrointestinal symptoms 40-60%

Urinary tract infection 30-60%

Treatment

Non opioid and opioid analgesics are the main drugs used to treat pain.

Antidepressants, anticonvulsants, and other CNS-active drugs may also be used for chronic or

neuropathic pain and are first-line therapy for some conditions. Neuraxial infusion, nerve

stimulation, injection therapies, and neural blockade can help selected patients. Cognitive-

behavioral interventions (eg, changes in relationships in the home; systematic use of relaxation

techniques, hypnosis, or biofeedback; graduated exercise) may reduce pain and pain-related

disability and help patients cope.

Non opioid analgesics

Acetaminophen and NSAIDs are often effective for mild to moderate pain (see Table: Non

opioid Analgesics). Of these, only ketorolac and diclofenac can be given parenterally. Non

opioids do not cause physical dependence or tolerance. Acetaminophen has no anti-

inflammatory or antiplatelet effects and does not cause gastric irritation.

NSAIDs include non selective COX (COX-1 and COX-2) inhibitors and selective COX-2

inhibitors (coxibs); all are effective analgesics. Aspirin is the least expensive but has

prolonged antiplatelet effects. Coxibs have lowest risk of ulcer formation and GI upset.

A 1.5% solution of diclofenac has been shown to effectively treat pain and limited joint

function caused by osteoarthritis of the knees; dose is 40 drops (1.2 ml) appliedqid to each

affected knee. Other topical diclofenac formulations that may be useful for local pain relief

include a patch (applied bid over the affected area) or a 1% gel (2 g qid for the upper

extremities or 4 g qid for the lower extremities).

Opioid Analgesics

“Opioid” is a generic term for natural or synthetic substances that bind to specific opioid

receptors in the CNS, producing an agonist action. Opioids are also called narcotics a term

originally used to refer to any psychoactive substance that induces sleep.

Opioid Analgesics Dose:

Opioid analgesics are useful in managing acute and chronic pain. They are sometimes

underused in patients with severe acute pain or with pain and a terminal disorder such as

cancer, resulting in needless pain and suffering. Reasons for under treatment include

Underestimation of the effective dose

Overestimation of the risk of adverse effects

Generally, opioids should not be withheld when treating acute, severe pain; however,

simultaneous treatment of the condition causing the pain usually limits the duration of

severe pain and the need for opioids to a few days or less.

Adverse effects

In opioid-naive patients, adverse effects common at the start of therapy include

Respiratory depression (rare with appropriate doses)

Sedation and mental clouding

Nausea and vomiting

Itching

Because steady-state plasma levels are not approached until 4 to 5 half-lives have

passed, drugs with a long half-life (particularly levorphanoland methadone) have a risk of

delayed toxicity as plasma levels rise. Modified-release opioids typically require several

days to approach steady-state levels.

In the elderly, opioids tend to have more adverse effects (commonly, constipation

and sedation or mental clouding). Falls are a particular risk in the elderly. Opioids may cause

urinary retention in men with benign prostatic hyperplasia.

AIM AND OBJECTIVE

AIM

The present study was mainly aim to investigate analgesic activity of aqueous

extract of tribulus terristries in analgesic induced and hot plat model in albino rats.

OBJECTIVE

This study was planned to examine.

Phytochemical screening.

Evaluation of anti analgesic activity,(aqueous extract of T.terristeries) in analgesic induced

and hot plate model.

Plan Work

Plant collection Photochemical studies Extraction of plant material activity

Analgesic activity Pharmacological activity Photochemical test activity

RIVEW AND LITERATURE

Literature Review on Activities of plant

Prabha et al., 2008.

T.terristites was investigated for analgesic and anti-inflammatory activities. The analgesic

activity was compared with aspirin by tail-immersion and acetic acid induced writhing

methods. The anti-inflammatory activity against carrageenan induced paw edema in rats was

compared with ibuprofen. In both the methods encouraging results were obtained. 

Hajera Khatun et al., 2011.

p.murax fruit extracts were investigated for analgesic and anti oxidant activity. analgesic

activity was examined using acetic acid-induced writhing test and formalin test in rat. Results

suggest that the fruit extract of p.murax possesses remarkable analgesic properties.

Hajera Khatun et al., 2011.

T.terristites. fruit extracts were investigated for antinociceptive and insecticidal& antifeedent

activities. Different anxiolytic, muscle relaxant, sedative-hypnotic drug actions were acting

through GABAA .

PLANT PROFILE Tribulus territites

Pl ant name : GokhruBotanical name : Tribulusterristris Synonyms : Puncture vine, TribulusFamily : Zygophyllaceae

Vernacular names Bengalli : Gokhru, GokuriHindi : Chota gokhru, Bara gokhruKannada : NegaluMalayalam : NeringilPunjabi : LotakSanskrit : Gokshur ,Brihat gokshurTamil : Nerunji, MullTelugu : Palleru

Taxonamy of plantClass : DicotyledonsSubclass : PolypetataeSeries : ThalamifloraeOrder : GeranialesFamily : ZygophyllaceaeGenus : TribulusSpecies : Terrestris

Botanical description

It is a creeper that is about 2 to 3 feet long having branches spread all over, leaves are in pairs of

5 to 8 and is of irregular shape. Flowers are small and yellow colored. Fruits are round and

possess 5 to 12 compartments and each compartment contains a seed. The seeds contain

aromatic oil. Roots are 4 to 5 inch long, brown in color and bear a sweet aroma. The plant

flowers in early winters followed by fruiting. P. murex L. is a succulent herb found near sea coast

of south India and some tropical areas of India. It appears during the month of July – September.

It grows luxuriously in fertile soils and crop land as a weed at temperatures of 25–30 degrees.

Chemical constituents

Fruit: Alkaloids 3.5%–5%, stable oil, aromatic oil, resins, glycosides, carbohydrates, saponins

and triterpenoids.

Stem: Saponins, herman, phytosterols, tannins and carbohydrates.

Root: Reducing sugars, phenolic compounds, saponins, xanthoproteins, alkaloids, triterpenoids

and flavonoids.

Leaves: Flavonoids, alkaloids, steroids, resins, saponins and proteins.

Phyto chemical constituents

Preliminary chemical examination of T. tersistris  revealed presence of naturally occurring

different chemical constituents. Whole plant is reported to contain medicinally important.

Mainly fruits contain alkaloids (3.5%–5%), stable oil, aromatic oil, resins, carbohydrates,

saponins, glycosides, and Triterpenoids and also two important flavonoids like 2′, 4′, 5′-

trihydroxy-5, 7-dimethoxy flavonoids and triacontanyl dotriacontanoate.Alkaloids,

steroids, resins, saponins and proteins are also reported.

Medicinal uses

Fruits are used as a diuretic, tonic and al so in the treatment of calculous affections and

painful micturition.

They are al so used as aphrodisiac &in gout.

Gokhru a common ingredient of Ayurvedic preparations, dashamoolarishta&chyavanprash.

The dried fruits of pendulum murex are known as Bara gokhru.

The fruits contain fatty oil, mucilage, an alkaloid &resin.

Bara gokhru is used as a diuretic and in dysuria&gonarrhoea.

Method of preparation of plant extraction

The fruits of the plant were collected in the month of January and dried in the shade.

The shade dried fruits are powdered separately to get coarse powder.

About 500g of dried and coarsely powdered fruits was extracted first with water by

continuous hot percolation using iodine flasks.

The extraction was carried out, by using solvents of increasing polarity starting from

water respectively.

The extraction was continued for 24 hrs.

The aqueous extract was filtered and concentrated to a dry at room temperature. a `brow

colour residue was obtained (8gm).

PHYTOCHEMICAL TESTS Various chemical test are performed on the extract of medicinal plant to identify the

phytochemical constitutions. Test for Alkaloids Dragendroffs test Wagner’s test Test for Carbohydrates Molish testTest for Tannins ferric chlorideTest for Flavonoids; Shindo Test Ferric chloride test:Proteins Heat test Test with trichloroacetic acid Hydrolysis test Biuret testTriterpenoids Libermann-Burchard Test

PHARMACOLOGICAL STUDIES

Analgesic activity

The anti analgesic activity of aqueous extract of tribulus terristates was in induced by

eddy’s hot plate and writhing response methods.

Animals

Swiss albino rats of both sexes weighing (150-200g) were obtained from venkateswara

trades; Bang lore & maintain in the animal house. They were properly housed in separate

cages in room temperature (25c) & fed with stanadard diet and water ad libitum .

Twenty four hours before each experiment (expect for the sub chronic toxity test), the

animals were deprived of food but not of water.

Eddy’s hot plate method

Swiss albino rats were divide into six groups, each of three animals .Group 1 served

as control ,Group 2 served as standard and were injected Diclofenac sodium

(9mg/kg)l intraperitonially Group 3.

And 4 were treated orally with aqueous extracts of 500 and 1000 mg/kg body weight

respectively.

Group 5 and 6 were treated orally with alcohol extracts of 500 and 1000 mg/ kg body weight

respectively

The animals were individually placed on the hot plate maintained at 55 c , one hour after their

respective treatments.

The response time was noted as the time at which animals reacted to the pain stimulus either by

paw licking or jump response, whichever appeared first the cut off time of the reaction was 15

seconds.

WRITHING RESPONSE

Acetic acid induced writhing method was adopted for evaluation of analgesic activity.

writhing is defined as a stretch, to one side, extension of hind legs, contraction of the abdomen so

that the abdomen of mice touches the floor turning of trunk (twist).

Any writhing is considered as a positive response.

Swiss albino mice weighing between 15-35g were used for evaluation of analgesic activity ;in

each group six albino mice were kept.

A solution of acetic acid (1%v/v) in distilled water was prepared. A solution of

ibuprofen (dose-100mg/kg/10ml) was prepared in normal normal saline water. Test – 1:A

solution of etoricoxib (5mg/kg) in combination with diclofenac potassium (10m/mg) was

prepared in 10ml of normal saline water.

Wister albino mice of either sex were divide into four different group each

containing six animals were marked individually. Food was withdrawn 12 hours prior to

drug admintered till completion of experiment. The animal were weighed and number

appropriately. The test and standard drugs were given orally. After 60min writhing was

induced by intraperitonal injection of 1% of acetic acid in volume of 0.1ml/10g body

weight .the writhing episodes were collected for 30 min stretching movements consisting of

arching of the back, elongated of body and extensions of hind limbs were coned.

RESULTSPRELIMINARY PHYTOCHEMICAL ANALYSIS The results of preliminary phytochemical analysis of aqueous extract are shown in table. Table: Data showing the phytochemical screening of fruit extracts of tribulus terristities.

S.NO CONSTITUENTS AQUEOUS EXTRACT

1 Carbohydrates +

2 Glycosides +

3 Alkaloids +

4 Saponins +

5 Stable oil +

6 Tannins +

7 Proteins +

8 Flavonoids +

9 Terpinoids +

10 Cardiac glycosides +

11 Fats _

“+” = Presence“_” = Absence

GROUP TRETMENT REACTION TIME IS SECONDS AT TIME (min)

0 30 60 90 120

GROUP I Control (saline po)3.41±0.06

3.52±0.04

3.61±0.03

3.56±0.08

3.68±0.07

GROUP II Standard(ibprofene)(100 mg/kg po)

3.21±0.03

7.31±0.04**

8.42±0.02**

9.52±0.09**

8.25±0.08**

GROUP III Aq. extract of T. terristities(100 mg/kg po)

3.36±0.04

3.65±0.05

3.75±0.08

3.85±0.06

3.95±0.09

GROUP IV Aq. Extract of T. terristities (200mg/gm Po)

3.54±0.05

5.56±0.07**

6.65±0.06**

7.35±0.09**

6.56±0.07**

GROUP V Aq.exatract of T.terristites(400gm/mg Po)

3.55±0.02

8.31±0.05**

9.25±0.07**

10.56±0.06**

9.52±0.07**

Table1: shows analgesic activity of aqueous fruit extract of Tribulus terristries by Eddy’s hot plate

method.

**P <0.01 significant with respect to control group. Values are expressed as mean ± S.E.M; N=6 in each group.

Statistical analysis one way AONOVA followed by dunnett,s test.

Table 2: shows analgesic activity of aqueous fruit extract of Tribulus terristris by writhing response method.

**P <0.01 significant with respect to control group. Values are expressed as mean ± S.E.M; N=6 in each group. Statistical analysis one- way ANOVA followed by Dunnett’s test.

Groups Tretment No.writhings in (10 mints) % of inhibition

Group I Control(saline po) 32.43±0.02 _

Group II Standrad(Ibuprofene 100mg/kg po)

18.13±0.04** 44.09**

Group III Aq. extract of T.terristities(100mg/kgPo)

30.12±0.06 7.12

Group IV Aq. extract of T.terristities(200mg/kg Po)

20.15±0.03** 37.86 **

Group V Aq. extract of T.terristities(400mg/kg Po)

16.16±0.01** 50.16**

DISCUSSION

Generally medicinal values of the plants are dictated by their phytochemical and other

chemical constituents. The phytochemical analysis aqueous extract of tribulus terristities

revealed the presence of major phyto constituents like flavonoids tannins, saponins,

terpinoids, alkaloids, glycosides & carbohydrates. Hence the result of present investigation

indicate that the test drug may possess pharmacological proterties like analgesic. Theses

result also commensurate with earlier reported phytochemicals of this plant.

The preliminary phytochemical study revealed the presence of alkaloids,

carbohydrates, tannins, flavonoids. It helps to undertake further studies on the isolation and

identification of specific phytoconstituents. Extract showed the analgesic activity when

compared with control and analyzed statistically by ANOVA comparison test. On the

basis of these findings, it may be inferred that Tribulus terristris. In discussion, this study

provides evidences for the analgesic activity of Tribulus terristris.

CONCLUSION

Herbal drug are the option of treatment of disease which carries less side- effect and

toxicity.

Preliminary phytochemical studies revealed that the presence of saponins, tannins,

terpinoids, glycosides, alkaloids, flavonoids, carbohydrates, stable oils treatment with

aqueous extract of Tribulus terristris at the dose of 400mg/kg showed a significant

analgesic activity by eddy’s hot plate method and writhing response method.

The results of present study indicates the aqueous fruit extract of Tribulus terristris.

Possesses analgesic effect, which is in accordance with its ethno medical use.

Analgesic effect of the extracts was demonstrated in the experimental models using

eddy’s hot plate and writhing response methods using thermal stimuli, an increase in

reaction time is generally considered an important parameter of analgesic activity.

Tribulus terristris shows significant results for analgesic activity. 

REFERENCES

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Distibutors..Dehradun. 1995 p;308.

3.Agharkar SP.Medicinal plants of Bombay Presidemcy. Scientific Publisher

Jodhpur,India,1991.4.Olajide OA, Awe SO, and makinde,J M,pharmacological studies on the leaf of P sidium

g.Sauvaire and Baccon. Column techniques of Steroidal Saponins in Tribuilus Terristris linn. 1978; pg no.251.

5. Sivarajan VV, Balachandran I. Ayurvedic drugs and their plant sources. Oxford and IBH, New Delhi, 1994, 155-157.

6. Kolammal M. Pharmacognosy of Ayurvedic Drugs. Series 1, No. 2, Dept. of Pharmacognosy, Govt. Ayurveda College Trivandrum, 1978- 57-5

9. Singh NP, Panda H. Medicinal herbs with their formulation. Daya Publishing House, New Delhi, India, 2005, 671.

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