1. INTRODUCTION:

Research in herbal medicine has recently been revolutionized with the identification of several botanical plants with established physiological effect and efficacy for clinical condition either alone or combination with pharmaceuticals. Ayurveda is a comprehensive natural health care system that organized in India more than 5000 year ago.

It is still widely used in India as a system of primary health care, and interest in it is growing worldwide as well. Ayurveda has unique concept and methodologies to address health care throughout the course of life, from pregnancy and infant care to geriatric disorder. Verities of common spices as well as herbs, herbal mixture .and spatial preparation known as rasayanas are utilized to modulate the disease physiology. Research has been conducted worldwide on ayurveda and results were very encouraging and strongly advocating it to the various treatments of various ailments such as depression, anxiety, sleep disorder, hypertension, diabetes mellitus, etc. ¹

A vast assay of illness and medicinal complaint has always been and always will be a real part of the human condition. Plants have supplied human with cases for their ailments from relieving headache to treating heart disease. Since the time of earliest human evolution, the impact of plant-derived medicine on human history has been remarkable. Opium, snake root, digitalis, fever bark and chaulmoogra have all left their leaf print on the human time line. These medicinal plant and others are the foundation and future of the human medicine. The vast natural pharmacy of the plant medicine is accounted for the 80% of the substances that used to cause the disease before use learned to synthesize medicinal compound in the laboratory.

Inflammation is a complex pathophysiological process mediated by a variety of singling molecules produced by leukocytes, macrophages and mast cells as well as by the activation of complement factors, which bring about edema formation as a result of extravasations of fluid, proteins etc and pain at the site of inflammation ². Despite the great numbers of non-steroidal anti-inflammatory drugs, their therapeutic efficacy seems to be hampered by the presence of a number of undesired, and often serious, side effects.³

Desmostachya bipinnata (L.) Stapf. is an indigenous medicinal plant of India and is commonly used by traditional healers for the treatment of inflammation, as anti obesity. so far various activity of Desmostachia bipinnata (L.) Stapf. there is no reported activity of the Desmostachia bipinnata (L.) Stapf. Clumps and rootstock, therefore the present study is to find out various activities which are claimed by traditional healer.

Herbal medicine is based upon the promise that plant contains natural substance that can promote health and alleviate illness. The WHO has estimated that perhaps 80% of the earth’s 6 billion people rely upon health care needs and major part of this therapy involves the use of plant extracts or their active constituents. Scientists in many part of their world have carried out extensive research and have proven to humanity, the effective use of herbal medicine. ⁴

About 130 pure chemical substances extracted from some 100 species of higher plants are used in medicines throughout the world. There is a massive health available in these collections. But what is coming in the way is lack of standardized products, lack of reliable production technology and absence of pharmacological profile of drugs. Allopathic system has recognized by the importance of herbal medicine and drugs used in their system are hormones including sex hormones, anti cancerous drugs, cvs drugs, etc. In development of herbal formulation like tablet, capsule, liquids etc. It is necessary of active of principle in formulation. ⁵

1.1 Inflammation:

Inflammation is a protective mechanism by the organism to remove the injurious stimuli and to initiate the healing process. Inflammation is a defense reaction in order to eliminate the spread of injurious agents. ⁶

The purpose of inflammation is to eliminate the initial cause of cell injury, clear out necrotic cells and tissues damaged from the inflammatory process, and to initiate tissue repair. ^{7, 8}

Type of Inflammation

(A) Acute inflammation

· Acute inflammation, usually of sudden onset, in which vascular and exudative processes predominate, does little damage.

· Immediate and early response to tissue injury.

(B) Chronic inflammation:

· A chronic inflammatory response happens over a long period of time.

· Chronic inflammation is caused by Long-term infection,Immune deregulations, Poor sleep, mental stress, Environmental toxins, Microorganisms, Fat cells.

Causes of inflammation:

(A) Physical agents

· Mechanic agents:

fractures, foreign corps, sand, etc.

· Thermal agents:

Burns, freezing

(B) Chemical agents:

Toxic gases, acids, bases

Biological agents:

Bacteria, viruses, parasites

Symptoms of Inflammation

i. There are four cardinal signs-

ii. Rubor (redness) caused by increased blood flow and pooling of blood.

iii. Calor (heat) due to increased blood flow.

iv. Tumor (swelling) because of edema and accumulation of inflammatory exudates clot formation in the effected tissue spaces.

v. Dolor (pain) is caused by chemicals such as kinin (especially bradykinin)

Pathophysiology of inflammation:

Inflammation is essentially a protective response intended to eliminate both the initial cause of cell injury (e.g. microbes or toxins) and the necrotic cell and tissues arising a consequence of such injury. ⁹

There are many causes of inflammation including food poisoning, infection, burns, radiations, and Inflammation may also be caused due to a chronic problem like viral the causes of the inflammatory response have the basic stages. Vasodilatation and increased permeability of blood vessels. Emigration of phagocytes from blood in to interstitial fluid.

In response to injury. ^{10, 11, 12}

i. Histamins: Mast cells, vasophyl, platelet, induced vasodialation.

ii. kinkns: this polypeptide formed from kinogens induced vasodialation.

iii. Prostaglandine: (PGs.) Lipid E, seriesare stimulates the vasodialation.

iv. Complement chemotaxsis

v. Cause of Inflammation

Bacterial Infections:

Several types of bacteria which get into our body through contaminated food or water are the main causes of inflammation.

Viral infections:

Many viruses are also responsible for the cause inflammation including rotavirus, Norwalk virus, cytomegalovirus, herpes simplex virus, and viral hepatitis.

Parasitic infections:

Is also a cause for inflammation.

Food Intolerance:

Some people are not able to digest some component of food properly, such as lactose, the sugar found in milk-which ultimately leads to inflammation.

1.2 Antiiflammatory Drugs^{13, 14}

Nonselective cox inhibitors (Traditional NSAIDs)

i. Salicylates: -Aspirin

ii. Propionic acid derivatives

· Ibuprofen

· Naproxen

· Ketoprofen

iii. Anthranilic acid derivatives

· Mefenamic acid

iv. Aryl-acetic acid derivatives

· Diclofinac,

· Aceclofenac

v. Oxicam derivatives

· Piroxicam,

· Tenoxicam

vi. Indole derivative

· Indomethacin

vii. Pyrazolone derivative

· Phenylbutazone

Preferential COX-2 inhibitors

· Nimesulide

· Meloxicam

Selective COX-2 inhibitors

· Celecoxic

· Etoricoxib

· Parecoxib

Table 1: List of Anti-inflammatory plants^{15, 16, 17}

Sr.No.	Botanical name	Common name	Uses
1.	Desmostachia bipinata	Kusha	anti-inflammatory and analgesic
2.	Cedrus deodara	Devedaru	Anti inflammatory and anti ulcer
3.	Sida cordifolia	Bala	Anti-rheumatic and anti-inflammatory
4.	Vitex negundo	Nirgundi	Anti-rheumatic, anti- inflammatory
5.	Commophora mukul	Guggulu	Anti-inflammatory, antirheumatic, hypolipidemic
6.	Glycyrrhiza glabra	Licorice (root)	anti- inflammatory
7.	Azadiracta indica	Neem (all parts)	anti- inflammatory, insepticidal
8.	Curcuma longa	Turmeric (root)	anti- inflammatory
9.	Gingiber officinale	Ginger (root)	anti- inflammatory

1.3 MECHANISM OF ACTION:

According to unifying concept of NSAID Drugs action during inflammation, pain, fever, Arechedonic acid librated from phspholipid fraction of the cell membrane, Arachidonic acid then converted via. Cyclooxygenase (COX-1 and COX-2) Pathways to prostaglandine (PGs). The step –I is oxidation of arachidonic acid in to endoper oxide, PGG2, and its subsiquents reduction to the hydroxyl, endoperoxide, PGH2, the letter is then transformed in to the primary prostanoids PGE2, PGF2, PGD2, PGI2, and TXA2. ¹⁸

Through COX-I and COX-2 are very similar. COX-1, Activity is constituently present in really all cell type at the constant level, COX-2, Activity normally absent from cell (except kidney and brain). thus COX-1is physiologically while COX-2 is pathologically. PGs sensitize blood vessel to the effect of the inflammatory mediators that increases permeability¹⁹. PGs. Particularly PGE2, PGF, hyperalgesia associated with inflammation They sensitize the chemical receptors of the affect the pain ending to other mediators such as bradykinin and histamine. Release PGs. in the CNS may lower the threshold of the central pain circuts.

The non selective NSAIDs drugs inhibits cox-1 7 cox-2 , while other act more selective on COX-2, Thus indomethacin, and piroxicam,10-40, fold selective for COX-1where as nabumetone, is 15 fold selectively for cox-2, ibuprofen and Asprin inhibits cox-1 and cox-2. ²⁰

1.4 PATHOPHYSIOLOGICAL MECHANISM:

Inflammation:

Inflammation was characterized two thousand years ago by Celsus by the four Latin words: Rubor, color, tumor and dolor. Inflammation has different phases: the first phase is caused by an increase of vascular permeability resulting in exudation of fluid from the blood into the interstitial space, the second one by infiltration of leukocytes from the blood into the tissues and the third one by granuloma formation. Accordingly, anti-inflammatory tests have to be divided into those measuring acute inflammation, sub acute inflammation and chronic repair processes. In some cases, the screening is directed to test compounds for local application. Predominantly, however, these studies are aimed to find new drugs against poly arthritis and other rheumatic diseases. Since the etiology of polyarthritis is considered to be largely immunologically, special tests have been developed to investigate various immunological and allergic factors. ^{21, 22}

1.5 Analgesic activity:

Pain is a symptom of many diseases requiring treatment with analgesics. Severe pain due to cancer metastases needs the use of strong analgesics that means opioid drugs. The addiction liability of opioids led to intensive research for compounds without this side effect. Many approaches have been used to differentiate the various actions of strong analgesics by developing animal models not only for analgesic activity but also for addiction liability. Several types of opioid receptors have been identified in the brain allowing in vitro binding tests. ²³ However, the in vitro tests can only partially substitute for animal experiments involving pain. Pain is a common phenomenon in all animals, at least in vertebral animals, similar to that felt by man. Analgesic effects in animals are comparable with the therapeutic effects in man. Needless to say, that in every instance painful stimuli to animals must be restricted.

As much as possible. Painful stimuli can consist of direct stimulation of the efferent sensory nerves or stimulation of pain receptors by various means such as heat or pressure. The role of endogenous peptides such as enkephalins and endorphins gives more insight into brain processes and the action of central analgesics. Pain can also be elicited by inflammation. Progress has been made in elucidating the role of various endogenous substances such as prostaglandins and peptides in the inflammatory process. Most of the so called non-steroidal anti-inflammatory agents have also analgesic activity. Lim and Guzman (1968) differentiated between antipyretic analgesics causing analgesia by blocking impulse generation at pain receptors in the periphery while the narcotic analgesics block synaptic transmission of impulses signaling pain in the central nervous system. Today, the classification into central and peripheral analgesics is definitively too simplified but provides a guide for differentiation by pharmacological methods. ²⁴

The pathophysiological mechanisms underlying the loss of intestinal fluid in inflammation have been the subject of much debate for decades inflammation may be caused by an increase in osmotic load within the intestine, excessive secretion of electrolytes and water into the intestinal lumen, exudation of protein and fluid from the mucosa, infection and inflammation; and altered intestinal motility, resulting in rapid transit. In most instances, multiple processes are simultaneously affected involving several factors, a particular factor becoming a dominant player in a given environment, however, motility and/or secretary disturbances usually remain a common denominator in most cases. The mucosal lining of the gastrointestinal tract is provided with an extensive nerve supply from the enteric nervous system. Neurotransmitters such as acetylcholine and noradrenaline and neurotransmitter candidates such as ATP, CGRP, CCK-8, ENK, GAL, GABA, serotonin, NO, somatostatin, SP, VIP etc have been implicated to different extents in normal and patho physiological situations. Based on the knowledge gained about the divergent factors controlling the processes of secretion of electrolytes and motility, many interventional strategies have been adopted by researchers and numerous anti diarrheal compounds have been developed but not many compounds are without side effects and therefore there has always been a need for finding new ones. ^25,
26

Acetylcholine is the endogenous neurotransmitter at cholinergic synapses in the central and peripheral nervous system. The stimulation of vagal input to the gastrointestinal tract increases tone, amplitude of contraction and secretory activity of the stomach and intestine. Since such responses are inconsistently seen with administered acetylcholine, possibly because of poor perfusion and rapid hydrolysis by plasma butryl cholinesterase, use of neostigmine was made in the present investigation. Neostigmine is an inhibitor of acetylcholinesterase and increases the amount of acetylcholine at the synapse and thus exerts a pro-kinetic effect. The results show that the Lantana camara leaf powder and LCME significantly reduced the % intestinal transit in a dose dependent manner. Lantadene A also produced a statistically significant reduction in % intestinal transit.

The induction of inflammation with Carragennin I results from the action of ricinoleic acid formed from hydrolysis of its triglyceride in the carragennin .The released ricinoleic acid produces changes in the transport of water and electrolytes resulting in a hypersecretory response and speeds intestinal transit. The involvement of nitric oxide from neurons in the diarrhea induced by the castor oil has also been proposed. Carragennin I increase the induction of prostaglandins, cause changes in the permeability and mucosal injuries and stimulate PAF biosynthesis which may result in inflammation of intestinal mucosa. The preventive administration of LCME was associated with significant protection against of inflammation with. Carragennin I in rats. Lantana camara might possess some compounds with anti secretory properties which may account for its efficacy against inflammation induced by carragennin in rats. ^{27, 28, 29}

Lantana camara has been reported to be toxic to grazing animals such as cow, buffaloes, sheep and goat and laboratory animals such as guinea pigs and female rats. In spite of its widespread toxicity in the Lantana affected animals, various parts of this plant have been used in the traditional medicines for treating cuts, ulcers, swelling, eczema, inflammation, fever etc. Gastrointestinal stasis, ruminal stasis, constipation, discolorization of conjunctiva, photosensitization, decreased bile flow and urinary retention in the Lantana poisoned animals has been noticed. These symptoms resembled those due to atropine toxicity i.e., anticholinergic excess. ³⁰

Anti-dysenteric and anti-inflammatory properties of medicinal plants have been suggested to be due to tannins, alkaloids, saponins, flavonoids, sterols and triterpenes and reducing sugars. The sesquiterpene lactones have been reported to have the ability to relax smooth muscles and thereby relieve. Result in decrease in motility. Although the anti-diarrheal properties of the reported active terpenoids are well established, aspects of their mechanism of action remain poorly understood. Terpenes, flavonoids and terpenoid derivatives may act by inhibiting release of autocoids and prostaglandins thereby inhibit the motility and secretion induced by neostigmine. Intestinal motility alterations in Lantana camara foliage poisoned sheep has been described by Pass et al. But no mechanism has been suggested. ³¹

2. INTRODUCTION TO PLANT ^{32, 33, 34}

PLANT PROFILE

Name: Desmostachia bipinnata (L.) Stapf.

SYNONYMS:

Hindi: Kusha

Ayurvedic: Kusha, Suchyagra, Yagyabhuushana, Kshurapatra.

Taxonomy

Kingdom : Planteae

Subkingdom : Tracheophyta

Superdivision : Spermophyta

Order : Poales/Cyperales

Family : Poaceae (grass family)

Sub family : Chloridideae

Genus : Desmostachya

Species : Desmostachia bipinnata (L.) Stapf.

Table 2: Some medicinally used plants, family poaceae³⁵

Parts Used	Biological Source	Action
Whole plant	Cynodon dactylon	Antiemetic.
Stem and root bark	Fragmites australis	Diuretics
Root	Cympopogon flexviosus	Astringent
Stem bark	Zea maize	Laxative
And root	Vetiveria zizinoides	urinary and skin disease
Root	Lemon grass	Bacteriostatic
Twing	Agrstis capillaries	Anti-rheumatism
Bark and latex	Periploca aphylla	Used in cerebral fever
Root	Achanatherum hymenoides	Purgative
Stem and root	Desmostachia bipinnata (L.) Stapf.	Antimicrobial

Figure No. 02 Root and clumps of (Desmostachia bipinnata (L.) Stapf.

Figure No. 03 Desmostachia bipinnata (L.) Stapf.Plant

TRADITIONAL AND MODERN USE:

The Ayurvedic Pharmacopoeia of India indicated the use of root and leaf in asthma and dyspnoea, stem bark in diseages of the spleen. Bhavaprakasa used Desmostachia bipinnata (L.) Stapf. in enlargement of spleen. Powder of the root bark is an excellent substitute for epecacunha in dysentery. ³⁶

Plant is used as a traditional medicinal plant with unique properties. Traditionally of Desmostachia bipinnata (L.) Stapf is used alone or with other medicinals to treat common disease such as fevers, rheumatism, indigestion, cough, cold, eczema, asthma, elephantiasis, nausea, vomiting. According to Ayurveda, dried whole plant is a good tonic, expectorant, depurative, and anthelmintic. The dried root bark is a substitute for ipecacuanha. The root bark is febrifuge, anthelmintic, depurative, expectorant, and laxative. The powdered root used in asthama, bronchitis, and dyspepsia. The leaves are useful in the treatment of paralysis, arthralegia, swellings, and intermittent fevers. The flowers are bitter, digestive, astringent, stomachic, anthelmintic, and tonic. It is also a reputed Homoeopathic drug. 37

THERAPEUTIC USES:

1. Plant pacifies vitiated pitta,

2. Diarrohea,

3. Dysentery,

4. Menorrhagia,

5. Antibacterial

6. Jaundice,

7. Skin disease,

8. Burning sensation,

9. Excessive perspirant,

10. Anti helicobacter pylori

11. Anti-ulcerogenic

12. Diuretic,

13. Calcium channel blocking activity.

MAJOR CHEMICAL CONSTITUENTS^38,39,40

Plant contain five main flavonoid glycosides were isolated, for the first time, from the ethanol extract of Desmostachia bipinnata (L.) Stapf. (Gramineae). They were identified as kaempferol, quercetin, quercetin-3-glucoside, trycin and trycin-7-glucoside.

The sample of essential oil was obtained from the aerial parts of the plant by hydrodistillation. From the 16 compounds representing 99.97% of the oils: camphene (16.79%), isobornyl acetate (9.92%), tricyclene (4.30%), (+,-) trans-2, 6-gamma-Irone (2.21%), Caryophyllene diepoxide (12.29%), β-eudesmol (11.16%) Eseroline (25.15%) and Calarene (3.48%) appear as the main components. Plant contains xanthenes also. The plant contains flavonoids and carbohydrates also.

Bioactive compound:

Plant contain five main flavonoid glycosides were isolated, for the first time, from the ethanol extract of Desmostachia bipinnata (L.) Stapf. (Gramineae). They were identified as kaempferol, quercetin, quercetin-3-glucoside, trycin and trycin-7-glucoside. Alkaloids. (such as hordenine, gramine and 5-methoxy –N-methyl tryptamine), flavonoides which have antioxidant activities and lower the concentration of blood triglycerides and cholesterol, Some bioactive compound such as–3–O–rutinoside, isorhamnetin-3–O–glucopyranoside , taraxasteryl acetate etc were isolated.

Plant and essential constituents:

A careful further study of plants within the Gramineae family that have similar properties may help reveal the manner in which certain groups of active constituents can explain the traditionally attributed herbal effects. Recently, a number of Gramineae plants has been utilized specifically for the treatment of immune- deficiency caused by cancer therapy (or other medical intervention, such as radiation therapy), these and other from this plants family have been used for different purpose. The mechanism of action has not been fully elucidated and comprehensive study of the chemical constituents and pharmacology is needed to determine whether these components from the same family of the plants share common structure function relationship. The leafy culms are used for thatching in India, and in India and Sudan, for roving into a coarse rope. In India the culms are considered diuretic and are used to treat dysentery and menorrhaeia. Plant contain five main flavonoid glycosides were isolated, for the first time, from the ethanol extract of Desmostachia bipinnata (L.) Stapf. (Gramineae). They were identified as kaempferol, quercetin, quercetin-3-glucoside, trycin, and trycin-7-glucoside. Alkaloids (such as hordenine, gramine and 5-methoxy –N-methyl tryptamine), flavonoides which have antioxidant activities and lower the concentration of blood triglycerides and cholesterol.

3. MATERIALS AND METHODS:

3.1 MATERIAL:

COLLECTION AND IDENTIFICATION:

The root and clums of Desmostachia bipinnata (L.) Stapf. Were collected from wild sources surrounding Lucknow, U.P.

3.2 METHOD:

Macroscopy:

Color : Green color

Odor : None

Taste : None

Texture : Short, rough.

Size : 2-3 m long

Microscopy:

A small portion of dried part of root put in hot water for 30 min. after that the T.S. of root with different staining supervision under microscope. Root shows a thick portion of rhytidoma, made of about 15-20 alternate bands of and dead cells of secondary cortex and secondary phloem, rectangular, about 4-6 layered, parenchymatous cells, filled with brown pigment. Secondary phloem consists of sieve elements, phloem fibers, fibers present and stone cells mostly rectangular in groups of 3-5 with bands of phloem fibers. ⁴¹

The Transverse section of root showed the fallowing characteristics:

i. Stele: These include pericycle, vasicular bundle, and pith.

ii. Pericycle: It is outer layer of stele consists of few layers of parenchymatous cells.

iii. Xylem and Phloem: The xylem and phloem are arranged in ring, The arrangement is radial, xylem is exarch.

iv. Trichome: Trichomes are absent but cuticle is present.

v. Epidermis: Epidermis is single layered present with stoma.

vi. Hypodermis: Hypodermis is sclerenchymatous cells, multilayered are present.

vii. Cortex: It is continues, narrow large parenchymatous zone, having oval/spherical cell, with intracellular space, containing brownish granular amorphous, store watery matter.

viii. Edodermis: This forms complete ring around stele .its cells are barrel shaped, they are thick due to mass of parechymatous cells.

ix. Vasicular bundle: Vasicular bundle are scattered in the ground tissue. Surrounded by bundle sheath. Vasicular bundle are colletertal closed, Vasicular bundle are usually oval shaped.

3.3 PHYSIOCHEMICAL INVESTIGATION:^42,43,44

Total ash value:

Weight the accurately 2-3gm of the air dried crude drug in a tarred platinum of silica dish and incinerate at a temperature not exceeding 450 ⁰ c until free from carbon, cool and weight. If a carbon free ash cannot be obtained in this way exhaust the charred mass with hot water, collect the residue on an ash less filter paper, incinerate the residue and filter paper until the ash is white or nearly so, add the filtrate, evaporate to dryness and ignite at a temperature not exceeding 450 ⁰C. Process repeated three times. Calculate the percentage of ash with the reference to the air dried drug.

Acid insoluble ash:

Boil the ash for five min. with 25 ml water, collected the insoluble matter in a gooch crucible are an ash less filter paper, Wash with hot water and ignite for 15 min. at a temperature not exceeding 450⁰C. Subtract the weight the insoluble matter from the weight of the ash, the difference in weight represent the water soluble ash.. Process repeated three times Calculate the percentage of water soluble ash with reference to the air dried drug.

Determination of moisture contents:

Loss and drying method:

The air dried root of Desmostachia bipinnata (L.) Stapf was determined by using the procedure.

Accurately weighed quantity of sample was taken in tared glass bottle and initial weight was taken .The sample was heated at 105⁰C in an oven and weighed. Process repeated three times, this procedure was repeated until a constant weight was obtained. The moisture content of the sample was calculated with reference to air dried drug.

3.4 PHYTOCHEMICAL INVESTIGATION^45,46,47,48

Methods of Extraction

EXTRACTIVE VALUE:

The water-soluble and alcohol soluble extractive values of air dried sample were evaluated using the procedure.

Succesive extraction:

100 gm of dried root clumps powder was extracted successively with various solvent in increasing order of polarity viz. n-hexane, ethyl- acetate, ethanol, and water. Each extract was concentrated to small volume and allowed to dry in dessicater. After drying, the respective extracts were weighed and percentage yields were calculated.

n-hexane extractive value

Macerate 5 grams of air dried drug with 100 ml n-hexane of the specified strength in a closed flask for twenty-hours, shaking frequently during six hours and allowing standing for eighteen hours, filtering it. Evaporate 25 ml of the filtrate to dryness in a tarred flat bottomed shallow dish and dry at 105⁰C to constant weight. Calculate the percentage of n-hexane soluble extractive with reference to air dried drug.

Ethyl-acetate extractive value:

Macerate 5 grams of air dried mark of the drug with 100 ml ethyl acetate of the specified strength in a closed flask for twenty-hours, shaking frequently during six hours and allowing standing for eighteen hours. Then filter it, evaporate 25 ml of the filtrate to dryness in a tarred flat bottomed shallow dish and dry at 105⁰c to constant weight and calculate the percentage of ethyl acetate soluble extractive with reference to air dried drug.

Water soluble extractive value:

Macerate 5 grams of air dried mark of the drug with 100 ml chloroform water of the specified strength in a closed flask for twenty-hours, shaking frequently during six hours and allowing standing for eighteen hours. And filter it. To evaporate 25 ml of the filtrate to dryness in a tared flat bottomed shallow dish and dry at 105⁰ c to constant weight and weigh. Calculate the percentage of water soluble extractive with reference to air dried drug.

Alcohol soluble extractive value:

Macerate 5 grams of air dried mark of the drug with 100 ml alcohol of the specified strength in a closed flask for twenty-hours, shaking frequently during six hours and allowing standing for eighteen hours. And filter it. To evaporate 25 ml of the filtrate to dryness in a tarred flat bottomed shallow dish and dry at 105⁰ c to constant weight and weigh. Calculate the percentage of alcohol soluble extractive with reference to air dried drug.

Hydro-alcoholic extraction:

250 gm of air dried powder was extracted with (50:50) water: ethanol by cold maceration in a specified strength in a closed flask for twenty-four hours, shaking frequently during six hours and allowing standing for eighteen hours. The extract was filtered, concentrated under reduced temperature using rotary evaporator and dried extracts was subjected for phytochemical, and biological activity on experimental animal model.

3.5 QUALITATIVE CHEMICAL ANALYSIS:

The various extract i.e. n-hexane; ethyl-acetate, ethanol, and water were subjected to qualitative chemical analysis to detect the presence of various phytocontituents. ^{49, 50}

Test for Carbohydrates

i. Molisch’s test:

Treat the test solution with few drops of alcoholic alpha-napthol. Add 0.2 ml of con. H₂SO₄ slowly through the sides of test tube, purple to violet color ring appears at the junction of two layers.

ii. Benedict’s test:

Treat the test solution with the few drops of Benedict’s reagents (alkaline solution containing cupric citrate complex) and upon boiling on water bath, reddish brown precipitate forms if reducing sugars are present.

iii. Fehling’s test:

Equal volume of Fehling’s A (copper sulphate in distilled water) and Fehling’s B (Potassium tartarate and sodium hydroxide in distilled water) reagents are mixed and few drops of sample is added and boiled. A brick red precipitate of cuprous oxide forms, if reducing sugars are present.

iv. Caramelisation:

Carbohydrates when treated with strong sulphuric acid, they undergo charring with the dehydration along with burning sugar smell.

v. Bromine water tests:

It gets decolorized by aldose but not by the ketose, because bromine water oxidizes selectively the aldehyde group to carboxylic group, giving raise to general class of compound called aldonic acid.

Test for Tannin:

i. Ferric chloride test: Test solution give blue-green color with Fecl_3.

ii. Vanillin hydrochloride Test: Test solution when treated with few drops of Vanillin hydrochloride reagents gives purplish red color.

iii. Alkaline reagents tests: Test solution with sodium hydroxide solution gives yellow to red with in short time.

iv. Tannins: Get precipitated in the solution when treated with heavy metals.

v. Tannins: Yields bulky precipitate with phenazone especially in the presence of sodium and phosphate.

Tests for Proteins and Amino acids:

i. Millon’s test: Test solution with 2 ml of millon’s reagents (Mercuric nitrate in nitric acids containing traces of nitrous acid) gives white precipitate, which turns red upon gentile heating.

ii. Ninhydrin test: Amino acid and proteins when boiled with 0.2% solution of Ninhydrin (Indane 1, 2, 3,-trione hydrate, Violet color appear.

Tests for Sterols and Triterpenoids:

i. Libermann-Burchard test:

Extract treated with few drops of acetic anhydride, boil and cool, concentrated sulphuric acid is added from the side of test tube, shows brown ring at the junction of two layers and upper layer turns green which shows the presence of sterols and formation of deep red color indicates the presence of triterpenoids.⁵¹

ii. Salkowski’s test:

Treat extract in chloroform with few drops of concentrated sulphuric acid, shake well and allow to stand for some time ,if red color appears at the lower layer indicates the presence of sterols and appearance of yellow colored at the lower layer is shows the presence of triterpinoids.

Tests for Glycosides:

General test for the presence of glycosides:⁵²

Test 1: Extract 200 mg of the drug by warming in a test tube with 5 ml of dilute 10% sulphuric acid on water bath at 1000c for two minutes, centrifuge or filter, pipette off supernatant or filtrate. Neutralize the acid extract with 5 % solution of sodium hydroxide (noting the volume of NaOH added). Add 0.1ml of Fehling’s solution and then B until alkaline (test with ph paper) and heat on water bath for 2 min. Note the quantity of red precipitate formed and compare with that formed in test 2.

Test 2: Extract 200mg of the drug using 5 ml of water instead of sulphuric acid .After the boiling add volume of water equal to the volume of NaOH used in the above test. Add 0.1 ml of Fehling A and B until alkaline and heat on water bath for two minutes. Note the quantity of red precipitate formed. Compare the precipitate formed in the test 2 with that formed in test 1. If the precipitate in test 1 is greater than in the test2 then Glycoside may be present. Since test 2 represent the amount of free reducing sugar already present in the crude drug, where as test 1 represent the free reducing sugar plus those released on acid hydrolysis of any glycosides in the crude drug.

Tests the extract for free Sugar⁵³:

After complete removal of free sugars, the extract is hydrolyzed with mineral acid and then tested for the glycone and aglycone moieties.

i. Raymond’s Test: The test solution when tested with dinitrobenzene in hot methanolic alkali gives violet color.

ii. Legal’sTest: Treat the extract with pyridine and add alkaline sodium nitroprusside solution, blood red color appears.

iii. Bromine water Test: Test solution when treated with bromine water gives yellow precipitate.

3.6 CHEMICAL TESTS FOR GLYCOSIDES^54,55

Tests for Glycosides:

A. Test for Glycosides:

i. Froth Test: Place 1ml solution of drug in water in semi-microtube shake well and note the stable froth.

ii. Haemolysis test:

Add 0.2 ml solution of saponin (prepared in1% normal saline) to 0.2 ml of v/v blood in normal saline and mix well, centrifuge and note the red supernatant. Compare with a control tube containing 0.2 ml of 10% blood in normal saline diluted with 0.2 ml of normal saline.

Test for Anthraquinone Glycosides:

i. Borntrager’s test:

Boil test solution with 1ml of dilute sulphuric acid in a test tube for 5min ( anthracene glycoside are hydrolyzed to aglycone and sugar by boiling with acids centrifuge or filter while hot (if centrifuged hot, the plant can be removed while anthacene aglycone are still sufficiently soluble in hot water, they are however insoluble in cold water),pipette out the supernatant or filtrate ,cool and shake with an equal volume of dichloromethane (the aglycon will dissolve preferably in dichloromethane). Separate the lower dichloromethane Layer and shake with half its volume with dilute ammonia. A rose pink color is produced in ammonical layer. (Aglycones based on anthraquinones give red in presence of alkali).

ii. Modified borntrager’s test:

Boil 200 mg of the test material with 2 ml of dilute sulphuric acid, 2ml of 5 % of aqueous ferric chloride solution for 5 min. and continue the test as above. As some plant contains anthracene aglycones in reduced form, if ferric chloride is used the extraction, oxidation to anthaquinines takes place, which shows response to the Borntrager’s test.

Tests for Cardiac Glycosides:

i. Kedde’s test:

Extract the drug with chloroform ,Evaporate to dryness, add one drop of 90 % of alcohol and 2 drops of 2 % 3,5 –dinitro benzoic acid (3,5-dinitrobenzene carboxylic acid-kedde’s reagent) in 90 % alcohol. Make alkaline with 20 % sodium hydroxide solution. A purple color is produced. The color reaction with 3, 5 dinitrobenzoic acids depends upon the presence of alpha, beta unsaturated-gamma lactones in the aglycone.

ii. Keller killiani test (test for Deoxy sugars):

Extract the drug with the chloroform and evaporate it to dryness. Add 0.4 ml of glacial acetic acid containing a trace amount of ferric chloride. Transfer it to small test tube; add carefully 0.5 ml of concentrated sulphuric acid by side of the test tube, blue color appears in acetic acid layer.

Test for Cyanogenic Glycosides:

Place 200 mg of drug in a conical flask and moisten with few drops of water. There should be no free liquid in the bottom of the flask (the tests will not work if there is any liquid in the flask as the hydrogen cyanide produced will dissolve in the water rather then, come off as a gas to react with the paper) Moisten a piece of picric acid paper with sodium carbonate solution (5 %aqueous) and Suspend the strip of sodium picrate paper by means of cork in the neck of the flask, warm gently at about 37 0c. Observe the change in color. Hydrogen cyanide is liberated from cyanogenatic glycosides by the enzyme activity and reacts with sodium picrate to form the reddish purple sodium isopicrate.

Tests for Alkaloids:

i. Mayer’s test: Alkaloids give cream color precipitate with Mayer’s reagent (potassium mercuric iodide solution)

ii. Dragendorff’s test: Alkaloids give reddish brown color precipitate with Dragendorff’s reagents (potassium bismuth iodide solution)

iii. Wagner’s test: Alkaloids give reddish brown precipitate with Wagner’s reagent.(Solution of Iodine in Potassium Iodide).

iv. Hager’s test: Alkaloids give yellow color precipitate with Hager’s reagent (Saturated solution of picric acid)

v. Tannic acid Test: Alkaloids give buff color precipitate with 10 % tannic acid solution.

Test for Flavonoids:

i. Shinoda test (Magnesium Hydrochloride reduction test): To the test solution add few fragments of magnesium ribbon and add concentrated hydrochloric drop wise, pink scarlet, crimson red or occasionally green to blue color appears after few minutes.

ii. Zinc-hydrochloride reduction test: To the test solution add a mixture of zinc dust and conc. Hydrochloride. It gives red color after few minutes.

iii. Alkaline reagent test: To the test solution add few drops of sodium hydroxide solution, formation of intense yellow color which turns to colorless on addition of few drops of dilute acetic acid indicates the presence of flavonoids.

3.7 CHROMATOGRAPHIC STUDIES^56,57,58

Thin layer chromatography (TLC) studies were carried out for various extracts to confirm their presence in the extracts.

TLC is mode of liquid chromatography in which extracts is applied as the small spot to the origin of a thin sorbet layer supported on the glass plate. The mobile phase move through stationary phase by capillary action some time assisted by gravity or pressure.TLC separation takes place in the open layer with each component having the same total migration time but different migration distance. Mobile phase consist of single or mixture of the solvents. Silica gel G used as stationary phase.

PROCEDURE:

Slurry of silica gel G was prepared in distilled water and poured over a glass plate to form a thin layer film. The prepared plates were allowed for setting (air-drying). After setting, the plates were kept in an oven at 100-120⁰c (30min) for activation (removal of water).The extracts was dissolved in alcohol or any other suitable solvent and spotted over an activated plate. It was kept in previously saturated developing chamber containing mobile phase and allowed to run 3/4 of the height of the plate. The developed plate was removed, air dried, sprayed with spring reagent and heated in an oven at110⁰c for about 15 min. for visualization of spots. The Rf. was calculated using following formula.

Distance traveled by the solute from the origin

Resolution

Factor (Rf.)= Distance traveled by the solvent front from the origin

3.8 PHYSIOLOGICAL INVESTIGATION⁵⁹:

Animal selection

Extracts used

Biological activity

Animal selection:

Healthy adult Wister albino rats were used for all the experimental study of various parameters of inflammatory activity And Analgesic activity. The body weight of these animals varied from 160 to 190 gm and the age from 4-6 months. Male or female rats were selected to anti-Inflammatory, Analgesic activity. Animals were housed individually with free access to food and water under standard condition and the basal food intake, body weights to the nearest gram were noted. The animals were starved 18 hr prior to starting biological activity.

Extracts used:

Total hydro alcoholic extract. The above extracts were selected n present study based upon the result obtained from the pilot study and the qualitative chemical analysis.

Biological activity:

Anti-inflammatory activity:

Carragenin method for paw oedema:

The method described by Awouters et al was followed. Healthy albino rats of the either sex (160-190 g) were divided into 5 groups of 6 animals each. They were fasted for 18 h prior to the test, with free access to water. Group I received the vehicle (01 ml of 0.1tween 80,( carragenin 0.1ml.induced edema.) and served as the control group. Groups II, III, IV and V were treated with standard drug (Indomethacin 5mg mg/kg s.c), hydro-alcoholic extract (200, 300mg, and 00mg/kg), respectively. All drugs/vehicle were administered orally (p. o.).

Thirty minutes after the drug treatment, each rat was administered 1 ml of tween-80 orally and housed separately in metabolic cages, with special provision to examine. The paw edema episodes were observed for 4 h. The cumulative height in plethismometer was noted (in ml.) at the end of the 4th h. Percent inhibition of inflammation was calculated using the mean height of mercury. Anti-inflammatory activity was determined term of percentage of edema the control group using the formula % inhibition = (1-Vt/ Vc) 100, Vt and Vc are the mean relative changes in the paw volume of the test and control group respectively. Data were analyzed by student’s t-test and the level of significant was set at p<0.001.

Schedule for screening of hydro-alcoholic extracts for anti inflammatory activity:

Group: Screening:

Control: 0.1ml 1% Tween 80 Saline.

Standard: 5 mg/kg. b.w. of Indomethacin in 2%Tween80 Saline.

DPE: 200mg/kg.b.w.in 2%Tween80 Saline.

DPE: 300mg/kg.b.w.in2%Tween80 Saline.

DPE: 400mg/kg.b.w.in 2%Tween80 Saline

% inhibition=(1-Vt/ Vc) 100, Vt and Vc.

3.9 ANALGESIC ACTIVITY^60,61,62:

Hot plate method for analgesia:

The method described by Awouters et al was followed. Healthy albino rats of the either sex (160-190 g) were divided into 5 groups of 6 animals each. They were fasted for 18 h prior to the test, with free access to water. Group I received the vehicle (01 ml of 0.1tween 80, orally, and served as the control group. The groups II, rats were administrated with standard drug Analgin at dose 150 mg\kg body weight. Orally, Group III, IV, V and VI were treated with, hydro-alcoholic extract (200, 300mg and400mg/kg), respectively. All drugs/vehicle were administered orally (p. o.).The reaction time was noted at 30, 60, 90, 120, and 180 min. of the interval of the drug administration.

Percent protection against paw licking was calculated using the following formula, % inhibition=(Wt-Wc/20-Wc) 100, Wt and WC are the mean value of the test and control groups, respectively. The data were analyzed by student; s t-testand the level of the significance was set at p<.001.

Figure No. 04 Hot plate

Schedule for screening of hydro-alcoholic extracts for analgesic activity:

Group: Screening:

Control: 0.1ml/kg b.w. Tween 80 Saline.

Standard: 150 mg//kg. b.w. of Analgin in 2% Tween80 Saline.

DPE: 200 mg/kg.b.w.in 2%Tween-80 Saline.

DPE: 400 mg/kg. b.w. in 2% Tween-80 Saline.

DPE*: 400mg/kg.b.w.in 2%Tween80 Saline.

% Protection=(wt-w c /20-WC) ×100

4. RESULT AND DISCUSSION:

4.1 RESULT:

4.1.1 PHARMACOGNOSTICAL INVESTIGATION:

Macroscopic:

Figure No.05 Root and clumps of (Desmostachia bipinnata (L.) Stapf. Plant.

Figure No. 06 Desmostachia bipinnata (L.) Stapf .Plant

Microscopic

Figure No. 07 T. S. of Root of Desmostachia bipinnata (L.) Stapf.

Figure No. 08 T.S. of clums of Desmostachia bipinnata (L.) Stapf.

4.1.2 PHYSIOCHEMICAL CONSTITUENTS:

Various physical constituents for roots and culms were determined which includes extractive values, moisture content, ash values.

Table 3: Physical constituents of plant Desmostachia bipinnata

Sl. no.	Parameter	Result in %w/w
1.	Total Ash value	3.5
2.	Acid Soluble ash	1.5
3.	Water Soluble ash	2.3
4.	Moisture Contents	11.0

4.1.3 PHYTOCHEMICAL INVESTIGATIONS:

Qualitative chemical examination of various extracts revealed by presence of flavonoids, tannins carbohydrates, triterpenoids, glycosides, alkaloids. Further thin layer chromatography study is carrying out to confirm the presence of above phytoconstituents.

Table 4: Percentage extractive and physical characteristic of various extracts of Desmostachia bipinnata (L.) Stapf.

Sl. no.	Extracts	% yield	Color	Odor	Consistency
1.	n-hexane	1.2	Yellow	Characteristic	Sticky
2.	Ethyl –acetate	3.3	Blackish/brown	Characteristic	Sticky
3.	Ethanol	5.5	Reddish Brown	Characteristic	Sticky
4.	Water	4.5	Reddish Brown	Characteristic	Sticky

4.1.4 PHYTOCHEMICAL TESTS OF SUCCESSIVE EXTRACTS:

Phytochemical investigation:

Table 5: Phytochemical investigation Desmostachia bipinnata (L.) Stapf.

Sl. No.	Phytoconstituents	n.Hexane	Ethylacetate	Ethanol	Water
1.	Alkaloids	-	+	+	+
2.	Flavonoids	-	-	+	+
3.	Glycosides	-	-	-	+
4.	Carbohydrates	-	+	-
5.	Proteins andAmino acids	-	-	-	+
6.	Tannins and Phenolic comp.	-	-	+	-
7	Sterols	+	-	+	+

4.1.5 TLC PROFILE OF VARIOUS SUCCESSIVE EXTRACTS:

n- HEXANE EXTRACT:

Table 6: TLC of Desmostachia bipinnata (L.) Stapf. in n- Hexane extract

Sl.N.

Solvent system

Spraying reagent

After spraying

Rf- value

Toluene: Ethyl acetate (93:7)

Con.H₂SO₄+vanillin

Blackish

0.42,

0.52,

0.64

ETHYL-ACETATE EXTRACT:

Table 7: TLC of Desmostachia bipinnata (L.) Stapf. In Ethyl-acetate extract

Sl. No.

Solvent system

Spraying reagent

After spraying

Rf value

Toluen: Ethylacetate

:Di-ethylamine

(70:20:10)

Methanolic Dragendorff’s reagent

Reddish

brown

0.28,

0.34,

0.22

Benzene:Glacial

Acetic acid

methanol (1:1:3)

Phenol+conH₂SO₄

Greenish

0.74,

0.54

AQUEOUS EXTRACT:

Table 8: Aqueous extract of Desmostachia bipinnata (L.) Stapf.

Sl. No.	Solvent system	Spraying reagent	After spraying	Rf value
1.	Toluene:Ethyl acetate:Diethyamine (70:20:10)	Dragendorffs reagent	Brownish yellow	0.52, 0.53
2.	Ethyl acetate:Formic acid:Glacial acetic acid:water(100:11:11:26)	Diphenyl benzyloxy ethylamine	Greenish black	0.64, 0.34
3.	Ethylacetate:Methanol: Water( 81:11:8)	Con.H₂SO₄, (heat1-3 minutes at 80^OC then spray)	Brown	0 .73, 0 .92

ALCOHOLIC EXTRACT:

Table 9: Alcoholic extract of Desmostachia bipinnata (L.) Stapf.

SL. No.

Solvent system

Spraying reagent

After spraying

Rf value

Toluene:Ethyl acetate:Diethyamine (70:20:10)

Dragendorff’s reagent

Blackish brown

0.73,

0.54

Ethyl acetate:Formic acid:Glacial acetic acid:water (100:11:11:26)

Di phenylbenzyloxy ethylamine

Greenish blue

0 .38,

0.69

4.1.5.1 TLC PLATE OF ALKALOIDS BY VARIOUS EXTRACTS

Figure No. 09 TLC plate of n-Hexane Extracts.

Figure No.10 TLC plate of Ethyl acetate extracts.

Figure No.11 TLC plate of Aqueous extracts

Figure No. 12 TLC plate of Alcoholic extracts

4.2 DISCUSSION:

In the present study, preliminary qualitative phytochemical tests revealed the presence of sterols, carbohydrates, flavonoids, triterpenes and saponins in the extracts of Desmostachya bipinnata (L.) Stapf. The inhibition of experimental anti-inflammatory and analgesic agent.substance are the basis of the pharmacological evaluation of a potential anti-inflammatory and analgesic agent. Many anti-inflammatory and analgesic agent acts by reducing the reduction of paw oedema. Inhibitors of prostaglandin biosynthesis delay castor oil-induced inflammation and pain. It has been shown that E type of prostaglandins cause inflammation in experimental animals as well as in human beings. The mechanism has been associated with dual effects on gastrointestinal motility as well as on water and electrolyte transport. PGE ₂also inhibits the absorption of glucose, a major stimulus to the intestinal absorption of water and electrolytes. The anti-inflammatory activity of the extracts was comparable to the standard drugs. The activity might be due to tannins and flavonoids present in these extracts. ⁶³

Study of the plants Desmostachya bipinnata (L.) Staff, we find out the following physiochemical property like total ash value (3.5), acid soluble ash (1.5), water soluble ash (2.3), moisture content (11.0).

Tannins, flavonoids, alkaloids, saponins, reducing sugars, sterols and triterpenes are reported for their anti-inflammatory activity. Tannins can evoke an antidiarrhoeal effect and these substances may precipitate proteins of the enterocytes, reduce peristaltic movement and intestinal secretion. ⁶⁴

The anti-inflammatory activity of flavonoids has been ascribed to their ability to inhibit intestinal motility and hydro-electrolytic secretion, which are known to be altered in this intestinal condition. In vitro and in vivo experiments have shown that flavonoids are able to inhibit the intestinal secretary response induced by prostaglandin EIn addition, flavonoids possess antioxidant properties which are presumed to be responsible for the inhibitory effects exerted upon several enzymes, including those involved in the arachidonic acid metabolism.

The Stem bark of the plant extracts (i.e. Hydro-alcoholic) of Desmostachya bipinnata (L.) Stapf. contain tannins, flavonoids, saponins, sterols and triterpenes, which could have contributed to the anti-inflammatory activity. The hydroalcohalic extracts were screened for anti-inflammatory activity using fallowing models and in each case the dose of the extracts administered was 200, 300 ,400 mg/kg body weight. ⁶⁵

In the microscopy of the plats we find out these properties like xylem and phloem are arranged in ring, the arrangement is radial, xylem is exarch. Trichomes are absent but cuticle is present. Epidermis is single layered present with stoma. Hypodermis is sclerenchymatous cells, multilayered are present. It is continuous, narrow large parenchymatous zone, having oval/ spherical cell, with intracellular space, containing brownish granular amorphous, store watery matter. This forms complete ring around stele. its cells are barrel shaped, they are thick due to mass of parechymatous cells. Vesicular bundle are scattered in the ground tissue. Surrounded by bundle sheath. Vesicular bundle are colletertal closed, Vesicular bundle are usually oval shaped. ⁶⁶

5. SUMMARY AND CONCLUSION:

The present study consists of Pharmacognostic, Phytochemical and Pharmacological screening of Desmostachya bipinnata (L.) Stapf. for anti-inflammatory activity. Pharmacognostic study of Desmostachya bipinnata (L.) Stapf.

Revealed the presence of root 2-5 alternate bands of cork and dead cells of secondary cortex and secondary phloem, Secondary cortex having stone cells, fibers, and prismatic crystal of calcium oxalate. Secondary phloem consists of sieve tube, fibers, phloem parenchyma. Phytochemical and chromatographic analysis of various extracts from stem bark of Desmostachya bipinnata (L.) Stapf. Indicate the presence of glycosides, alkaloids, flavonoids, carbohydrates, proteins and amino acids and sterols. ⁶⁷

The anti inflammation and analgesic effect of the extract may, thus, be attributable in inhibitory action against Carrageenin and Hot plate method for inflammation and pain. Flavanoids are known to reduced secretion and make the intestinal mucus resistant through the formation of protein Flavanoids. It may be possible that the tannins found to be Present in the extract could contribute to the observed effect. The antimicrobial activity of the extract of this plant has been reported. We conclude that Desmostachya bipinnata (L.) Stapf. has beneficial activities against inflammation and pain. ⁶⁸

6. FUTURE SCOPE OF THE WORK:

Analgesic is a major health problem especially for children under the age of 5 years and up to 17% of all death in the indoor pediatric patients is related to analgesic. Worldwide incidence of analgesic death account for more than 5-8 million each year in infants and small children less than 5 year especially in developing countries. According to WHO estimate for the year 1998, there were about 7.1 million deaths due to analgesic.

A range of medicinal plants with anti-diarrhoeal properties has been widely used by the traditional healers; however, the effectiveness of many of these analgesic traditional medicines has not been scientifically evaluated.

During the past decade, traditional systems of medicine have become a topic of global importance. Current estimates suggest that, in many developing countries, a large proportion of populations rely heavily on traditional practices and medicinal plants may be available in these countries, herbal medicines (phytomedicines) have often maintained popularity for historical and cultural reasons.

The Ayurvedic Pharmacopoeia of India indicated the use of root and leaf in asthma and dyspnoea, stem bark in diseages of the spleen. Bhavaprakasa used Desmostachia bipinnata (L.) Stapf. In enlargement of spleen. Powder of the root bark is an excellent Substitute for epecacunha indysentery. ⁶⁹

Desmostachia bipinnata (L.) Stapf. Is used as a traditional medicinal plant with unique properties. Traditionally calotropis is used alone or with other medicinals to treat common disease such as fevers, rheumatism, indigestion, cough, cold, eczema, asthma, elephantiasis, nausea, vomiting, and Pain killer.

According to Ayurveda, dried whole plant is a good tonic, expectorant, depurative, and anthelmintic. The dried root bark is a substitute for ipecacuanha. The root bark is febrifuge, anthelmintic, depurative, expectorant, and laxative. The powdered root used in asthama, bronchitis, and dyspepsia.

The leaves are useful in the treatment of paralysis, arthralegia, swellings, and intermittent fevers. The flowers are bitter, digestive, astringent, stomachic, anthelmintic, and tonic.

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Received on 14.08.2018 Accepted on 22.09.2018

Asian J. Pharm. Tech. 2018; 8 (4):211-226.

DOI: 10.5958/2231-5713.2018.00034.X

Treatment	Dose mg/kg b.w.	0min.	60min.	120min.	180min.	%Inhibition, Paw vol.
Control+Carragenin	0.1ml/mg	0.2±0.00	0.38±0.037	0.94±0.098	1.38±0.049	-----
Indomethacin	0.5	0.22±0.44	0.28±0.020	0.40±0.032	0.54±0.024	64%
DPE	200	0.22±0.44^b	0.28±0.20	0.42±0.020^b	0.76±0.040	46%
DPE	300	0.20±00	0.56±0.024^a	0.64±0.040	0.70±0.032^d	33.3%
DPE	400	0.20±00^a	0.26±0.024	0.46±0.024^c	0.58±0.020	62.5%

Treatment	Dose mg/kg b.w.	0min.	30min.	60min.	90min.	120min.	180min.	%Analgesia at 90 min
Control	0.1 ml/mg	2.62±0.20	3.13±0.13	2.5±0.20	2.4±0.24	2.75±0.32	2.62±0.38	----
Analgin	150	3.5±0.20	9.5±0.29	14.5±0.29	10.2±0.32	4.87±0.31	4.37±0.24	53%
DPE	200	2.62±0.45	5.0±0.20	5.37±0.24	6.36±0.24^b	5.0±0.35	5.00±0.35	24%
DPE	400	2.75±0.32	5.63±0.24	7.7±0.32	7.75±0.32^c	4.5±0.29^c	2.70±1.3	25%
DPE*	400	3.13±0.24^a	7.7±0.32	11±0.35	9.25±0.14	4.5±0.29	4.50±0.29^d	42%