ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. Calotropis Gigantea Linn. - A Complete Busket Of Indian Traditional Medicine S SARKAR 1*, R CHAKRAVERTY2, A GHOSH3. 1 Department of Pharmaceutical Chemistry, Durgapur, West Bengal, India 2 Department of Pharmacology, Durgapur, West Bengal, India. 3 Department of Pharmaceutics, Durgapur, West Bengal, India. Bengal College of Pharmaceutical Sciences and Research, B.R.B. Basu Sarani, Bidhannagar, Durgapur713212, W.B, India. *Corresponding author: SIPRA SARKAR, Email id: [email protected] -------------------------------------------------------------------------------------------------------------------------Abstract Calotropis genera comprise of two species, with 90% inhabiting southern Asian country and are most endemic to the India, Indonesia, Malaysia, Thailand, and Srilanka, China. Calotropis gigantea is a weed plant commonly known as giant milk weed. The plant is belonging to Apocynaceae family which includes latex bearing plants. C. gigantea is known for various medicinal properties in traditional medicinal system and use to cure a variety of diseases. In last few decades, C. gigantea is extensively studied for its medicinal properties by advanced scientific techniques and a variety of bioactive compounds have been isolated from the different parts of the plant and were analysed pharmacologically. The plant is reported for analgesic activity, antimicrobial activity, antioxidant activity, anti-pyretic activity, insecticidal activity, cytotoxicity activity, hepatoprotective activity, pregnancy interceptive properties, purgative properties, procoagulant activity and wound healing activity. The medicinal properties of this plant represent it as a valuable source of medicinal compound. This study is collective information concerning the ethnobotany, pharmacology, phytochemistry and biological activities of the C. gigantea. Keywords: Calotropis gigantea, ethnobotany, phytochemistry, pharmacological activity, future prospect. Introduction Arka (Calotropis giganteaa) an important drug of Ayurveda is known in this country from the earliest time. It is mentioned by the earliest Hindu writers and the ancient name of the plant which occurs in the Vedic literature was Arka alluding to the form of leaves, which was used in the sacrifical rites. There are two common species of Calotropis, viz. Calotropis gigantea (Linn.) R.Br. and Calotropis procera (Ait.) R.Br described by the Sanskrit writers. [1] C. gigantea is a common wasteland weed and commonly known as giant milk weed. This plant is a native of Bangladesh, Burma, China, India, Indonesia, Malaysia, Pakistan, Philippines, Thailand and Sri Lanka. C. gigantea is frequently available in India and used for several medication purposes in traditional medicinal system. [2] Most recently C. gigantea is scientifically reported for several medicinal properties (Figure 1) viz. the flowers are reported to possess analgesic activity , antimicrobial and 7 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. cytotoxic activity [3]. Leaves and areal parts of the plant are reported for anti-diarrhoeal activity [4], anti-Candida activity [5] and antibacterial activity [6], antioxidant activity.[ 7] Roots are reported to contain anti-pyretic activity[ 8], cytotoxic activity [9]. DESCRIPTION OF THE PLANT Taxonomical classification [10]:Kingdom: Planatae Subkingdom: Tracheobionta Superdivision: Spermatophyta Division: Magnoliophyta Class: Dicotyledones Sub class: Asteridae Series: Bicarpellatae Order: Gentianales Family: Apocynaceae Subfamily: Asclepidiaceae Genus: Calotropis Species: Calotropis gigantea Vernacular Names [11]:Common names: Giant Milkweed, Crown Flower, Swallow Wort. Hindi: Safed aak, Aak, Alarkh, Madar, Sveta Arka, Akanda, Bara Akand. Gujarati: Aakando English: Crown flower, giant Indian milkweed. Bowstring hemp, crownplant, madarMalaysia: Remiga, rembega, kemengu. Indonesia: Bidhuri (Sundanese, Madurese), sidaguri (Javanese), rubik (Aceh). Philippines: Kapal-kapal (Tagalog). Laos: Kok may, dok kap, dok hak. Thailand: Po thuean, paan thuean (northern), rak (central). Vietnam: B[oot]ng b[oot]ng, l[as] hen, nam t[it] b[at]. French: Faux arbre de soie, mercure vegetal. BOTANICAL DESCRIPTION Morphology of Calotropis gigantea leaf twig with oppositely arranged subsessile leaves; Broadly ovate or elliptical, cottony,pubescent when young and glabrous on maturity; Portion of the lamina showing venation pattern]Calotropis gigantea occurs as a single or many stemmed soft-wooded shrub, and occasionally a tree reaching to 6m. All parts of the plant exude white milky latex when cut. Botanical description of Calotropis incudes following parts: Bark & Branches The bark is thick, rough and corky and a yellow-brown colour; twigs are green and fleshy and may have a covering of tomentum (white fur like hairs)[Figure 1]. Leaves Leaves are opposite-decussate, simple,ovate to obovate with 4-6 pairs of subopposite nerves prominent on the abaxial surface, an acute apex, sessile (almost decurrent) base, a pale green colour, and quite large which is about 30x25 cm[Figure 1] . Inflorescences Inflorescences arise from the base of the leaves in pedunculate (c.7cm) cymes of 3-20[Figure 2]. Flowers Flowers consist of 5 small triangular dirty white sepals, 5 thick ovate petals (c1cm x 1cm)which are white at the base and purple at the tips and 5 purple tipped stamens, which surround a white 5 lobed stigma 11[Figure 2]. Fruits Fruits consist of green, spongy ovoid fruits (follicles), up to 15cm long by 10cm wide. They 8 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. split open to release plumed, papery light brown seeds with a pappus of white filaments up to 6cm long on one side. The main flowering period would be from March to October[Figure 2]. [12] Figure 1 Bark, leaves & Flowers Figure2 Fruits MACROSCOPICAL CHARACTERISTICS Macroscopical characteristics of various parts of Calotropis are as follows: Root The root occurs in the entire condition. The bark is separated from the wood 0.5-2.0 cm. in diameter bearing rootlets with diameter varying from 0.2 to 0.5 cm. externally whitish grey in colour, wrinkled in the fresh condition, plenty of whitish latex exudes from cuts or wounds in the bark. Fracture is incomplete. Leaf Simple, opposite, sub-sessile, slightly thick, fleshy, coriacious,10-15 cm. long and 4.5 to 6.5 cm. broad, broadly cuneate, obovate or obovate oblong, slightly cordate and auricled at base with tuff of short simple hairs on the upper side near place of the attachment to the petiole. The tender leaves are covered with ashy gray pubescence. Mature leaves are nearly smooth or even glabrous and pale green[Figure 1] Flowers Regular,bisexual, liliac or pale rose, purple or light greenish yellow and have a faint odour. They are arranged in simple or rarely compound cymose corymbs at the ends of laterally placed or interpetiolar peduncles arising from alternate sides of the nodes. Each cluster is surrounded by an involucre of several small oblong pointed scaly caducous bracts. Flower buds ovoid [Figure 2]. Calyx Five lobes broadly ovate with small fleshy teeth like glands within the base. Corolla Regular, gamopetalous, pale rose purple or liliac, subcordate to broadly sub- campanulate with a short tube and five broad ovate, lanceolate, valvate, spreading lobes. Stamens Five, inserted at the base of the coro Filaments united to form a large stamina column provided with five conspicuous radiating coronal appendages that are completely adnate to, but slightly shorter than the column. The appendages are fleshy, pale 9 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. purplish or yellowish white and laterally compressed with a circinnately recurved hollow corsal spur at base and two short obtuse obliquely divergent cuticles towards the top just below the apex. Anthers short, broad,somewhat horny with broadly triangular membranous anther tips that are inflexed over the sides of the stigmatic hood. Root bark The tap roots are found to be having prominent tops with rounded head and rest of the portion spirally curved. These hard roots are greyish white in colour and exhibit sap exudations at the places where bark has been cut. The bark of the older roots is cracked at places. The bark is yellowish grey outside and yellowish white inside. The upper cork portion is spongy and rough while the inner portion of bark is smooth and mucilaginous. The dried bark is bitter to taste . MICROSCOPICAL CHARACTERISTICS Microscopical characteristics of various parts of Calotropis are as follows: Stem (I) Epidermis: This is an outermost layer of uniseriate cells with thick cuticle. Uni- and multicellular hairs clothe epidermis almost completely. Cells are barrel to rectangular and are compactly arranged. (II) Cortex: These form a few layers below the epidermis which are collenchymatous (thickened corners). A few chloroplasts may also occur in these cells. Rest of the cortex is parenchymatous. Intercellular spaces are numerous. (III) Endodermis: This layer of uniseriate cells forms a wavy ring around the vascular tissue(separates cortex from underlying tissues.) The ells are barrel- rectangular shaped and are compactly arranged. Characteristic casparian thickening is lacking. It, however, contain starch grains (termed as starch sheath). (IV) Pericycle: It is in the form of small patches of sclerenchymatous fibres. A few parenchymatous cells of the original pericycle are present between these groups. (V) Vascular tissue system: Secondary growth is prominent. It shows groups of primary phloem, secondary phloem, cambium,secondary xylem, primary xylem and intraxylary phloem. Primary phloem is completely obliterated. Patches of secondary phloem occur above and close to the cambium.Cambium is unistratose.( but its derivatives on either side which are alike, give an appearance of a broad zone of cambium) . Secondary xylem forms a broad and extensive region. It compres vessels and tracheids. The annual rings are feeble. Primary xylem occurs near the pith and is endarch. A few groups of phloem are situated just below the primary xylem in the region of pith and are the groups of intraxylary or internal phloem. (VI) Pith: Centre is occupied by thin walled parenchyma and also many latex vessels. (VII) Points of ecological interest: A well differentiated cortex, presence of conjoint, bicollateral, opens and endarch vascular bundles indicate that the material is a dicotyledonous stem. Intraxylary phloem which is primary phloem of the bicollateral vascular bundle is characteristic. Leaf Transverse sections through the midrib showed an upper and lower, single- layered epidermis that was externally covered with a thick, striated cuticle, a few epidermal cells on both lower and upper surfaces, parenchymatous cells that were 10 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. thin-walled and isodiametric to circular. Intracellular spaces were present in ground tissue and the stele was crescent-shaped and composed of bicollateral and open vascular bundles. The xylem consisted mostly of vessels and tracheids, and a strip of cambium was present between the xylem and phloem tissues.The lamina which was dorsiventral with the mesophyll, was seen to be differentiated into a palisade and spongy tissue. The upper and lower epidermise were covered externally with a thick, striated cuticle. Below the upper epidermis were three rows of elongated,closely arranged, palisade parenchyma.Spongy parenchyma tissues were almost radially elongated with intracellular spaces.Central cells were irregular in shape; laticifersand vascular bundles were also present scattered in this region.[13] CHEMICAL CONSTITUENTS Phytochemical studies on Calotropis have afforded several types of compounds such as Cardenolide, triterpinoids,alkaloids, resins, anthocyanins and proteolytic enzymes in latex, flavonoids, tannins, sterol ,saponins, cardiac glycosides. Flowers contain -terpenes, multiflorenol, and cyclisadol . [14] Leaves The leaves contain mainly the amyrin,amyrin acetate, ß-sitosterol, urosolic acid, cardenolides, calotropin, calotropagenin. Latex The latex contains caoutchouc, calotropin,calotoxin 0.15%, calactin 0.15%, uscharin 0.45%, trypsin, voruscharin, uzarigenin,syriogenin and proceroside.[15] Flower The flower contains the flavonoids,queretin3- ratinoside, sterol, calactin, calotoxin, calotropagenin, calotropin, polysaccharides with D-arabinose, glucose, glucosamine and L-rhamnose. Flowers also contain enzymes 3-proteinase and calotropain(protease). Other chemical constituents of C. gigantea flowers are lupeol, uscharin,proceroside, proceragenin (cardenolide),syriogenin, taraxast-20(30)-en-3-(4methyl-3-pentenoate), 3-thiazoline cardenolide, gigantin,giganteol, isogiganteol, uscharidin, uzarigeninvoruscharin a-calotropeol, 3epimoretenol, alactuceryl acetate an a-lactuceryl isovalerate [16] Bark Root bark of Calotropis contains triterpenes, a new norditerpenyl ester, named Calotropterpenyl ester, and two unknown pentacyclic triterpinoids, namely calotropursenyl acetate and calotropfriedelenyl acetate, akundarol isovalerate, mundarol isovalerate and quercetin -3- rutinoside. [17,18] Propagation and management Propagation methods The tree seeds freely, and natural regeneration is common. Vegetative propagation through half stumps assumes a special importance as compared with the entire stumps because they help in faster multiplication of the parent genotype with plus characters, as each plant gives rise to 2 half stumps. Stumps also help in propagating only one plant. Vegetative propagation through stem and root cuttings is very useful in large-scale multiplication of the superior genotypes. Tree Management C. gigantea has been cultivated in South America and on the Caribbean Islands for the 11 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. production of fibres at a spacing of 1-1.5 m.When cultivated, annual yields of up to 500 kg/ha are expected. A single harvest per season is preferable to double(ortriple)harvest; a single harvest would result in a net saving of energy input both on the farm and in the processing plant. Well suited for intensive energy farming in arid or semiarid regions where frost is not a limiting factor.[19] PHARMACOLOGICAL ACTIVITY Antimicrobial activity Aqueous, methanol, ethanol and petroleum ether extracts of the leaves of C. gigantea were reported to possess anti-Candida activity against clinical isolate of Candida albicans, C. parapsilosis, C.tropicalis and C. krusei.[20] The aqueous extract of leaves of C. gigantea was reported to possess antibacterial activity against Staphylococcus aureus, Escherichia coli, Bacillus cereus, Pseudomonas aeruginosa, Micrococcus luteus and Klebsella pneumonia. [21] The aqueous extract of the latex of C. gigantea was reported to exhibit significantly inhibitory effect on S. aureus, B. cereus, E. coli and C. krusei. [22] Antifungal activity of C. gigantea was reported against plant pathogenic fungi like Fusarium mangiferae, that causes serious threat in mango cultivation. [23] Alam et al. (2008) reported the antibacterial activity of methanol extract from the root bark of C. gigantea and its petroleum ether, chloroform and ethyl acetate fractions. Both of methanol extract and its chloroform fraction showed activity against Sarcina lutea, B. megaterium and P. aeruginosa. Petroleum ether fraction showed activity against B. subtilis and Shigella sonnei, whereas ethyl acetate fraction showed activity against P. Aaeruginosa and E. coli.[24] Analgesic activity The alcoholic extract of the flowers of C. gigantea was reported for analgesic activity in chemical and thermal models in mice. The analgesic activity was performed by acetic acid induced writhing test and hot plate method. Oral dose of ethanolic extract of C. gigantea flower produced a significant decrease in the number of writhings and delay in paw licking time. [25] The CNS activity (analgesic activity) of alcoholic extract of peeled roots of C. gigantea was tested in albino rats. Analgesic activity was observed in Eddy’s hot plate method and acetic acid induced writhings. Oral dose of the extract (250 and 500 mg/kg body weight) significantly delayed the paw licking time and the numbers of writhings were greatly reduced. [26] Wound healing activity Root bark extract of C. gigantea was investigated for wound healing activity in Wistar albino rats. The rats were topically treated with extract formulated in ointment for excision wound healing models and extract was given orally (100, 200 and 400 mg/kg dose) for incision wound healing models. The results indicate that extract treatment accelerated wound healing in rats. [27] The crude latex of C. gigantea was evaluated for its wound healing activity in albino rats 12 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. using excision and incision wound models. At a dose of 200 mg/kg/day C. gigantea latex showed the significant wound healing activity as treated animals exhibit 83.42 % reduction in wound area when compared to controls which was 76.22 %. The extract treated wounds are found to epithelize faster as compared to controls. [28] Cytotoxic activity The cardenolide glycosides collected from the root C. gigantea were reported to carry cytotoxic activity against several human and mouse cell lines. Calotropin, frugoside and 4'–O-βDglucopyransylfrugoside was found as the active principles.[29] Two compounds (compound 1 and 2) isolated from ethanol extract of the roots of C. gigantea were reported to display inhibitory effects towards chronic myelogenous leukemia K562 and human gastric cancer SGC-7901 cell lines.[30] Crude ethyl acetate extract from the flower of C. gigantea was reported to inhibit the Ehrlich’s ascites carcinoma in mice. Intraperitoneal injection (50, 100 and 200 mg/kg body weight) of the extract significantly decreases the viable tumour cells and body weight gain induced by the tumour burden and prolonged survival time. The extract also restores the haematological and biochemical parameters (glucose, cholesterol, triglyceride, blood urea, ALP, SGPT and SGOT) that was altered during tumour progression, at 200 mg/kg body weight dose extract exhibits the best activity. [31] Anti-diarrhoeal activity The hydroalcoholic (50:50) extract of aerial part of C. gigantea was studied for anti-diarrhoeal activity against castor oil-induced-diarrhoea model in rats. The extract exhibited significant reductions in fecal output and frequency of droppings at the doses of 200 and 400 mg/kg body weight (intraperitoneal dose). The extract also showed significant inhibition in weight and volume of intestinal content. [32] Anti-pyretic activity Chitme et al. (2005) reported the anti-pyretic activity of the water:ethanol (50:50) extract of C.gigantea roots. Anti-pyretic activity was studied by using yeast and TAB (Typhoid) vaccineinduced pyrexia in Albino Swiss rats and rabbits. At the dose of 200 and 400 mg/kg body weight (intraperitoneal injection) extract significantly reduced the fever and body temperature was normalized. [33] Insecticidal activity Methanol extract of C. gigantea root bark and its chloroform and petroleum ether fractions were evaluated for residual film toxicity, fumigant toxicity and repellent effect against several inster of larvae and adult of Tribolium castaneum. Methanol extract showed high insecticidal activity against T. castaneum followed by petroleum ether fraction and chloroform fraction. None of the sample showed fumigant toxicity. [34] Anti-inflammatory Ethanol extract of C. gigantea was reported for the anti-inflammatory activity against carrageenan induced paw edema in Wistar albino rats. The oral administration of 400mg/kg of C. gigantea showed significant anti-inflammatory activity, the activity was found more than that of 100mg/kg of Ibuprofen. [35] Antioxidant activity 13 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. Leaves of C. gigantea were reported to carry antioxidant activity. The study reports the DPPH radical scavenging activity, reducing power activity and nitric oxide scavenging activity of the hydroalcohlic extract of C. gigantea leaves. Extract exhibited the maximum DPPH radical scavenging activity (85.17%) at 400μg/ml concentration. At 100μg/ml concentration extract showed 54.55% nitric oxide scavenging activity. Reducing power of the extract was found to increase with increasing the concentration of extract. [36] Pregnancy interceptive properties Different organic solvents of C. gigantea roots were reported to exhibit pregnancy interceptive activity in rats. The extract exhibited 100% pregnancy interceptive activity at a dose of 100 mg/kg.The extract also exhibited 100% efficacy at the dose of 12.5 mg/kg when administered in the Days 1-5 and 1-7 postcoitum schedules. [37] Procoagulant activity The latex of C. gigantea is reported to carry procoagulant activity. The latex extract hydrolysed casein, human fibrinogen and crude fibrin clot in a dose dependent manner. Extract hydrolyses the subunits of fibrinogen, subunit Aa hydrolyzed first followed by Bb and g subunit. The crude extract hydrolysis crude fibrin clot strongly compared to trypsin and papain. Proteins present in the latex of C. gigantea are strongly proteolytic and responsible for procoagulant activity of C. gigantea. [38] Hepatoprotective effects Ethanol extract of stems of C. gigantea was reported for hepatoprotective activity in male Wistar rats against carbon tetrachloride induced liver damage. The extract resulted in significantly decreased of AST, ALT and lipid peroxide levels and showed effective protection of liver. The extract also protects the rats from oxidative damage. [39] Vasodilation Effect: Effect of latex from Calotropis gigantea in the green frog R hexadactyla showed a significant increase in cardiac output. Evidence suggests the prime action of latex on the cardiovascular system involves changes in the cation (Ca, Na) permeability, with consequent excitation of Ca channels in the heart muscle and an increase coronary flow. Therefore, dilatation property is likely responsible for the pharmacologic actions of the latex.[40] SOME FORMULATION OF THIS PLANT Abedi et al published that Production of primary carbon has involved by the carbonization of Calotropis Gigantea (Giant Stabragh)in a negligibly ventilated atmosphere to drive out Comparison of K2co3 and Khco3 for Preparation of volatiles, leaving a porous carbon structure with low Carbonaceous Adsorbent. [41] Vidya C et al published that green synthesis of ZnO nanoparticles by zinc nitrate and utilizing the bio components of leaves extract of Calotropis Gigantea. The ZnO nano crystallites of average size range of 30-35 nm have been synthesized by rapid, simple and ecofriendly method. Zinc nanoparticles were characterized using scanning electron microscopy (SEM) and X-ray diffraction (XRD). [42] Conclusion Empirical knowledge about medicinal plantsplays a vital role in primary health care and has great potential for the discovery of new herbal drugs.The pharmacognostical studies including 14 ISSN: 2348 –0882 ============================================================================= Int. J. Pharm. Res. Sci., 2014, 02(1), 7-17. macroscopic and microscopic evaluation of various parts of Calotropis gigantea would be of considerable use in the identification of this drug. These findings may be useful to supplement existing information with regard to the identification and standardization of Calotropis gigantea to distinguish it from substitutes and adulterants. [43] In recent years, ethnomedicinal studies received much attention as this brings to light the numerous little known and unknown medicinal virtues especially of plant origin. Pharmacological screenings of C. gigantea revealed its medicinal potential and represents as a valuable medicinal plant with several medicinal properties. As the pharmacologists are looking forward to develop new drugs from natural sources, development of modern drugs from C. gigantea can be emphasized for the control of various diseases. A systemic research and development work should be undertaken for the conservation of C. gigantea and development of products for their better economic and therapeutic utilization. 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