Rajani Kanta Sahu et al. IRJP 2011, 2 (12), 243-248

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY ISSN 2230 – 8407 Available online www.irjponline.com Research Article

EVALUATION OF ANTIOXIDANT ACTIVITY IN ETHANOLIC EXTRACTS OF FIVE SPECIES Gayatri Nahak and Rajani Kanta Sahu* B.J.B Autonomous College, Bhubaneswar, ,

Article Received on: 14/10/11 Revised on: 25/11/11 Approved for publication: 09/12/11

*Email: [email protected]

ABSTRACT The , the largest family in the , comprises nearly 50 genera and 1,000 species and is pantropical, concentrated mainly in the old world, chiefly in Indo- malaysia. Members of the family yield spices, dyes, perfumes, medicines and a number of ornamental species are cultivated for their showy flowers. Recently this has acquired great importance in the present-day world with its antiaging, anticancer, anti-alzheimer’s diseases, antioxidant, and a variety of other medicinal properties due to its significant potential. The present study aims at comparing the antioxidant activity of five Curcuma species namely Curcuma longa, Curcuma zedoaria, Curcuma angustifolia, Curcuma aromatica and Curcuma amada based on their curcumin and phenol content. In our studying C.longa exhibited the highest antioxidant activity 74.61±0.02% at IC50 value 24µg/ml followed by C.zedoaria (63.27±0.06%), C.angustifolia (58.35±0.06%), C.aromatica (55.38±0.06%) and C.amada (52.61±0.02%). Antioxidant activity in four species except C.angustifolia has strong correlation with curcumin and phenol content. However C.angustifolia may be active due to high aromatic oil content like eugenol, palmitic and camphor etc. The natural oxides of curcuma species can be explained in the field of pharmaceutical areas for their uses in modern health care as phytoprotectants. KEY WORDS: Curcuma longa, Curcuma zedoaria, Curcuma angustifolia, Curcuma aromatica, Curcuma amada, Antioxidant activity, Phenol content.

INTRODUCTION chronic gonorrhoea, bruises, small pox, chicken pox, scorpion snake Zingiberaceae family constitutes a vital group of rhizomatous and leech bites, congestions, scabies, dyspepsia, ring worm, etc. medicinal and aromatic characterized by the presence of C.aromatica Salisb yields arrow root. Quite recently C.longa is volatile oils and oleoresins of export value. Generally, the proved to contain a chemical constituent called turmerin which is and fruits are aromatic, tonic and stimulant; occasionally they are anticarcinogenic, antipyretic and antiallergic. The importance of nutritive. Some are used as food as they contain in large has significantly increased due to its usefulness as quantities while others yield an astringent and diaphoretic juice. The pesticide, fungicide and bactericide51. important genera coming under Zingiberaceae is Curcuma. Turmeric Curcuma longa Linn. syn. Curcuma domestica Valeton. is a is a medicinal plant extensively used in Ayurveda, Unani and Siddha perennial herb, 60-90cm in height, with a short stem and tufts of medicine as home remedy for various diseases4,13. C.longa L., erect . is cylindric, ovoid, orange coloured and botanically related to ginger (Zingiberaceae family), is a perennial branched. Leaves are simple, very large, petiole as long as the blade, plant having a short stem with large oblong leaves and bears ovate, oblong-lanceolate, tapering to the base up to 45cm long. Flowers are pyri form or oblong rhizomes, which are often branched and pale yellow, arranged in spikes concealed by the sheathing petioles brownish yellow in colour. It also is used as a food additive (spice), and flowering are pale green51. The botanical description is preservative and colouring agent in Asian countries, including China also done by Gamble15, Kirtikar and Basu27, Sivarajan and and South East Asia. It is also considered as auspicious and is a part Balachandran40 and Thakur46. C.longa contains curcumin, alkaloid of religious rituals. In old Hindu medicine, it is extensively used for and an essential oil. Dry rhizomes of C.longa yield 5.8% essential the treatment of sprains and swelling caused by injury. In recent oil. A ketone and an alcohol are obtained from the volatile distillate. times, traditional Indian medicine uses turmeric powder for the Fresh rhizomes yield 0.24% oil containing Zingiberine11. The treatment of biliary disorders, anorexia, coryza, cough, diabetic colouring principle of turmeric is the main component of this plant wounds, hepatic disorders, rheumatism and sinusitis5. In India, and is responsible for the antiinflammatory property. Curcumin ladies anoint their bodies with turmeric paste, which is considered to (diferuloylmethane), the main yellow bioactive component of be an antiseptic. Purseglove36,37 states that it is a condiment and a turmeric has been shown to have a wide spectrum of biological spice among the rice eating peoples of South East Asia and actions. These include its anti-inflammatory, antioxidant, Indochina. Turmeric dyes is used in combination with some alkalies anticarcinogenic, antimutagenic, anticoagulant, antifertility, to colour silk and cotton. It is also used in several countries as a antidiabetic, antibacterial, antifungal, antiprotozoal, antiviral, colouring material in pharmacy, confectionery and food industries. It antifibrotic, antivenom, antiulcer, hypotensive and is an essential and sacred ingredient of all social, cultural and hypocholesteremic activities45,21,51. It’s antiinflammatory, anticancer religious functions and rites in India, especially in the South51. and antioxidant roles may be clinically exploited to control Aiyar1, Kirtikar and Basu27, Watt55, Haines18 and Holtumn20 describe rheumatism, carcinogenesis and oxidative stress-related the various uses of Curcuma. Purseglove36,37 also gives a limited pathogenesis. The successive extraction of C.longa with petroleum account of its uses. The medicinal properties of turmeric are ether, alcohol and distilled water yielded extracts when administered innumerable and very ancient. Kirtikar and Basu27 state that the on 1-7 days of pregnancy at dose levels of 100and 200mg/kg have rhizome is very pungent, bitter, healing, laxative, anthelmentic, been found to exhibit significant anti-fertility activity14. vulnerary, tonic, alexeteric and emollient. It is used as a medicine in Curcuma aromatica Salisb. is a perennial tuberous herb with various kapha and vata diseases of blood. In Comboidia it is used as annulate, aromatic yellow rhizome which is internally orange-red in a tonic and antipyretic. In China it is used as a stimulant, aspirant, colour. Leaves are elliptic or lanceolate-oblong, caudate-acuminate, carminative, cordeal, emmenagogue, astringent, detergent, diuretic 30-60cm long, petioles as long or even longer, bracts ovate, and matrient. It is used in Unani systems in treating, jaundice, recurved, more or less tinged with red or pink. Flowers are pink, lip scabies and bruises. On the whole it is used in the treatment of yellow, obovate, deflexed, sub-entire or obscurely three lobed. Fruits bronchitis, dropsy, vertigo, skin diseases, liver infections, burns, are dehiscent, globose, 3-valved capsules51. The plant is also boils, elephantiasis, sprains, hysteric effects, fevers, swellings, described by Gamble15 and Kirtikar and Basu27. Rhizomes yield

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011 Rajani Kanta Sahu et al. IRJP 2011, 2 (12), 243-248

6.1% essential oil11. Essential oil contains a-and b-curcumene, d- cause molecular transformations and gene mutations that curcumin camphene and p-methoxy cinnamic acid. The colouring matter is and other antioxidant products from the dried rhizome of turmeric curcumin. Rhizomes are used in combination with astringents and may be useful in the prevention or treatment of some age-related aromatics for bruises, sprains, hiccough, bronchitis, cough, degenerative processes31. This study was undertaken to verify the leucoderma and skin eruptions51. The rhizomes have an agreeable variation of total curcumin contents and antioxidant activity in fragrant smell and yield a yellow colouring matter like turmeric, and turmeric rhizomes collected from different villages of Koraput the fresh root has a camphoraceous odour. The dried rhizome is used district of Odisha. as a carminative and aromatic adjunctant to other medicines32. Oil is MATERIALS AND METHODS used for treatment of early stage of cervix cancer3. Verghese49 Plant Materials established the use of mango-ginger as an exotic flavourant. The rhizomes of five curcuma species i.e. Curcuma longa, Curcuma Curcuma zedoaria (Berg.) Rosc. syn. C.zerumbet Roxb; Amomum zedoaria, Curcuma angustifolia, Curcuma aromatica and Curcuma zedoaria Christm. has 4 -6 leaves with 20-60cm long lamina. The amada of family Zingiberaceae were collected from the local tribal lamina is oblong-lanceolate, finely acuminate and glabrous on people of Jeypore, Baipariguda, Sasahandi, Koraput, Orissa. Fresh both the surfaces. Flower stalk is 20-25cm long, emerging before the rhizomes were rinsed severally with clean tap water to make it dust leaves. Flowers are yellow, while the flowering is green tinged and debris free. Then they were dried in the shady condition for 3to4 with red. Calyx is 8mm long, corolla tube is twice as long as the days until they become moisture free. Dried rhizomes were ground calyx. Capsule is ovoid, trigonous, thin smooth and bursting in electric chopper to get fine powder form for further use. irregularly. Tubers are palmately branched and camphoraceous46. Preparation of plant extracts The botanical description of the plant is also given by Sivarajan and The dried and powdered form of rhizomes of curcuma species (each Balachandrann40. The medicinal uses of C.zedoaria were 50g) were extracted successively with ethanol (each 400ml.) for 10- summarized from various sources by Duke12 as antipyretic, 12 hrs, using a Soxhlet apparatus. Then collected solutions were aromatic, carminative, demulcent, expectorant, stomachic, stimulant, filtered through Whatman No-1 filter paper. The extracts were and tonic. A decoction of rhizomes along with long pepper, evaporated to dryness under reduced pressure at 900C by Rotary cinnamon, and honey is said to be beneficial for colds, fevers, vacuum evaporator to obtain the respective extracts and stored in a bronchitis, and coughs. Rhizomes are used in medicines given to freeze condition at −180C until used for further analysis. women after childbirth46. It is an odoriferous ingredient of the Phenolic Estimation cosmetics used for the chronic skin diseases caused by impure or The total phenol content of plant extracts were determined by using deranged blood32. Folin-Ciocalteu Spectrophotometric method according to the method Curcuma angustifolia (Zingiberaceae) is an attractive ginger with described26. Reading samples on a UV-vis spectrophotometer at 650 stout underground rhizomes which is also recognized as East Indian nm. Results were expressed as Catechol equivalents (mg/mg). arrowroot. In early spring the flowers are produced before the Estimation of Curcumin Contents leaves. Very colorful bracts make this a showy species. The Curcumin contents were determined from rhizomes of five Curcuma inflorences lasts in full bloom on the plants for about three weeks species i.e. Curcuma longa, Curcuma zedoaria, Curcuma and more. Leaves grow to about 2ft tall and die down in autumn. angustifolia, Curcuma aromatica and Curcuma amada by solvent This species is found in the Eastern Himalays and inhabits bright extraction and spectrophotometer method39. open hillsides woods7. Rhizomes are dried and powdered, and the Antioxidative activity starch obtained forms the chief source of Indian arrowroot. It is The evaluation of radical scavenging activity (antioxidant activity) nutritive and is used as an agreeable, non-irritating diet in certain was conducted by the method of (Brand-Williams et al.8 with chronic diseases, during convalescence from fevers, in irritations of modifications. The following concentrations of extracts were the alimentary canal, pulmonary organs, or of the urinary apparatus prepared 40μg/mL, 80μg/mL, 120μg/mL, 160μg/mL and 200μg/mL. and also used in consumption, excessive thirst, jaundice, kidney A stock solution of the sample (100mg/ml) was diluted for 5 disorder, and fattening the body38. The rhizomes are used in concentrations. Each concentration was tested in triplicate. The inflammation, bone fracture, intestinal diseases, etc. by the tribales portion of sample solution (0.5ml) was mixed with 3.0ml of 0.1mM of and Chattisgarh states of India23. 1,1-Diphenyl-2-2picrylhydrazyl (DPPH, in 95% distilled ethanol) Curcuma amada Roxb. is a rhizomatous aromatic herb with a leafy and allowed to stand at room temperature for 30 minute under light tuft and 60-90cm in height. Leaves are long, petiolate, oblong- protection. The absorbance was measured at 517nm.The scavenging lanceolate, tapering at both ends, glabrous and green on both sides. activity of the samples at corresponded intensity of quenching Flowers are white or pale yellow, arranged in spikes in the centre of DPPH. Lower the absorbance of the reaction mixture indicates tuft of the leaves. Lip is semi-elliptic, yellow, 3-lobbed with the mid higher free radical scavenging activity. The different in absorbance lobe emarginate51. The plant is also described by Gamble15and between the test and the control (DPPH in ethanol) was calculated Kirtikar and Basu27.The rhizomes of this plant are useful in vitiated and expressed as (%) scavenging of DPPH radical. The capability to conditions of pitta, anorexia, dyspepsia, flatulence, colic, bruises, scavenge the DPPH radical was calculated by using the following wounds, chronic ulcers, skin diseases, pruritus, fever, constipations, equation. strangury, hiccough, cough, bronchitis, sprains, gout, halitosis, Scavenging effect (%) = (1-As/Ac) ×100 otalgia and inflammations51. The fresh root possesses the smell of As is the absorbance of the sample at t =0 min. green mango and hence the name mango ginger. The rhizomes are Ac is the absorbance of the control at t=30 min. used externally in the form of paste as an application for bruises and In the DPPH test, antioxidants were typically characterized by their skin diseases generally combined with other medicines. Tubers IC50 value (Inhibition Concentration of Sample required to scavenge rubbed with the leaf-juice of Caesalpinia bonduc is given for 50% of DPPH radicals). The results were obtained by linear worms32. Rhizome is also used in applications over contusions and regression analysis of the dose response curve plotted using % sprains11. inhibition and concentration. Free radicals were a major interest for early physicists and RESULTS AND DISCUSSION radiologists and much later, the free radicals were found to be a Initially crude extracts were obtained through the extraction process product of normal metabolisms. Today, it is well known that radicals using ethanol as solvent and the amount of extracts yields C.longa INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011 Rajani Kanta Sahu et al. IRJP 2011, 2 (12), 243-248

(3.35gm), C.aromatica (2.56gm), C.zedoaria (2.67gm), C.amada methane, and Bis (4-hydroxy cinnamoyl) methame which shows (2.20gm) and C.angusifolia (2.62gm) respectively. The variation in 50% inhibhitory concentrations and IC50 values of these compounds yield may be due to the polarity of the solvents used in the extraction were lower than that of α-tocopherol meaning that they are more process (Table-2). A medicinal herb can be viewed as a synthetic effective antioxidants. Noguchi et al.33 reported that curcumin laboratory as it produces and contains a number chemical reacted with stable radicals such as galvinoxyl and N,N-diphenyl-l- compounds. These compounds are responsible for medicinal picryl hydrazyl, suggesting that it can serve as a hydrogen donor and activities are secondary metabolites. Qualitative analysis of five is a strong antioxidant as compared to eugenol. Song et al.43 reported curcuma species reveals the presence of alkaloids, saponins, free radical scavenging and hepato-protective activity of turmeric flavonoids, phenols, curcumin and anthraquinones (Table-1). The rhizomes in in vitro system, and the activity was much better than total phenolics in the extracts were determined that of ascorbic acid. The mechanism of antioxidative activity of spectrophotometrically by the Folin-ciocalteu method and the results curcumin has been reported by Masuda et al.30. He found that were reported as Catechol equivalents (mg/g). The highest curcumin formed dimers as radical termination products especially concentration of total phenol was 285mg/g present in the ethanolic at 2´-position of curcumin molecule and oxidative coupling reaction extract of C.longa rhizome followed by C.zedoaria (205mg/ml), at 3´-position29. Watanabe and Fukai52,53 have reported that C.aromatica(195mg/ml), C.angusifolia (110mg/ml) and curcumin suppresses the oxidative stress by scavenging various free C.amada(50mg/ml) respectively. The total phenolic content varied radicals andits antioxidative activity seems to be derived from its significantly between the five species of Curcuma. suppressive effects. Asai and Miyazawa2 showed that phenolic The unique properties of the spice are attributable to the essential oil yellowish pigments of turmeric display antioxidative activity in rats and curcuminoid components present therein. Curcumin possess while Okada et al.34 in his study on induced oxidative renal damage excellent antioxidant properties. It is reported to be more potent in in male mice showed that curcumin is an effective protectant against preventing lipid peroxidation than alpha-tocopherol, pine bark oxidative stress. Curcumin, having antioxidative property, may act extract, grape extract or the commonly used synthetic as anticancer agent, but also inhibits the regulatory enzymes and antioxidant BHT28,44. A complex of the three curcuminoids was exhibits anticarcinogenic action. A peculiar thing is noticeable in found to be more effective as an antioxidant than each of the case of C.angustifolia crude extract which contain a very negligible components-curcumin, demethoxycurcumin, or amount of curcumin content but having good antioxidant potential in bisdemethoxycurcumin-used alone. The results obtained from these comparison to C.aromatica and C.amada. It may be due to presence six curcuma species from Table-2 showed that the curcumin content of more than 30 components from the fresh and dried rhizome oils in C.longa exhibited the highest i.e. 8.22(mg/100mg) followed by of C.angustifolia of Vietnamese origin in which camphor (12.6 and C.zedoaria 7.35(mg/100mg), C.aromatica 6.07(mg/100mg) and 12.1%) and curzerenone (>57 and 38%) are the major constituents55. C.amada 1.54(mg/100mg) respectively where as C.angusifolia very It is interesting to note that a significant amount of methyl eugenol negligible amount of curcumin. Curcumin content is reported to vary (10.5%), α-muurolene (1.9%), pentadecanoic acid (1.8%), (E,E)- from one species to another. Several studies have shown that soil farnesyl acetone (1.3%) and n-heptyl salicylate (1.1%) are present factors, including nutrients and level acidity as well as the genus in the oil of C.angustifolia. Apart from the above, Nguyen et al.56 diversity, may affect the content of curcumin in plants 10,16. Similar also reported furanodienone and isofuranodienone from the dried results are obtained by Chavalittumrong and Jirawattanapon (1992) rhizome of C.angustifolia Roxb. who studied variation in the active constituents of C.domestica We observed a correlation between the DPPH radical scavenging rhizomes collected from Nakhon Pathom, Central Thailand, where activity of the plant extracts and their phenol contents with the they found the highest curcumin content was 10.12% w/w. In sample correlation coefficient R2= 0.861. A significant correlation is addition35, reported that a sample from the South contained the also seen between curcumin content and antioxidant activity of these highest total curcumin (8.99–0.83%w/w), while the lowest was Curcuma species with correlation coefficient R2= 0.735(Fig-3). The found in the North (4.80–1.83%w/w) where the climate is cooler and results showed that the antioxidant activity in turmeric was due to the dry period is longer and more pronounced. The significance of the presence of total phenol content. Consequently the antioxidant turmeric in medicine has increased considerably with the discovery activity of plant extracts are often explained with respect to their of the antioxidant properties of naturally occurring phenolic total curcumin content16,25. In a study by Majeed28, the free-radical compounds present therein. Curcumin exhibits strong antioxidant scavenging ability of various curcuminoids were evaluated using the activity6,17,4142,48. In DPPH assay, DPPH is a stable free radical with DPPH (1.1 diphenyl-2-picrylhydrazyl) radical scavenging method. purple color. The antioxidants scavenge DPPH radical by donating The results indicated that curcuminoids neutralize free radicals in a hydrogen atoms leading to a non-radical with yellow color. Table-3 dose-dependent manner. Tetrahydrocurcumin (THC) was the most and Fig-2 show the results of the free radical (DPPH) scavenging effective, followed by curcumin and bisdemethoxycurcumin. It is activity in (%) inhibition in all five Curcuma species. The result suggested that the antioxidant mechanism of curcuminoids may revealed that the ethanolic fraction of C.longa rhizome exhibited the include one or more of the following interactions: (1) scavenging or highest radical scavenging activity with 74.61±0.02% followed neutralizing of free radicals, (2) interacting with oxidative cascade C.zedoaria (63.27±0.06%) C.angusifolia (58.35±0.06%), C. and preventing its outcome, (3) oxygen quenching and making it less aromatic (55.38±0.06%) and C.amada (52.61±0.02%) respectively. available for oxidative reactions, (4) inhibition of oxidative enzymes Antioxidant activity using DPPH radical scavenging assay reported like cytochrome P-450, and (5) chelating or disarming oxidative 29 with IC50 value is shown in the (Table-2). The lower the IC50 is the properties of metal ions like iron (Fe). Masuda et al. investigated higher the antioxidant activity of the compound. The result showed the antioxidant mechanism of curcumin against peroxide radicals in the lowest IC50 value in ethanolic extract of C.longa which is very the presence of ethyl linoleate as one of the polyunsaturated lipids. close to ascorbic acid i.e. 24µg/ml followed by C. zedoaria They found that during the antioxidation process, curcumin reacted (40µg/ml), C. angusifolia (100µg/ml), C.aromatica (120µg/ml) and with four types of linoleate peroxyl radicals to yield six reaction C.amada (121µg/ml) respectively. Similar results are observed in products. On the basis of the formation pathway for their chemical C.longa by Ungphaiboon et al.47 and showed strong activity. The structures, an antioxidant mechanism of curcumin in antioxidant principle of turmeric as reported by Hirasa and polyunsaturated lipids was proposed, which involved an oxidative Takemasa19 includes curcumin, 4-hydroxy cinnmoyl (feruloyl) coupling reaction at the 3′-position of the curcumin with the lipid INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011 Rajani Kanta Sahu et al. IRJP 2011, 2 (12), 243-248 and a subsequent intramolecular Diels-Alder reaction. The 22. Iqbal M, Sharma SD, Okazaki Y, Fujisawa M, and Okada S. Dietary significance of turmeric in medicine has increased considerably with supplementation of curcumin enhances antioxidant and phase II metabolizing enzymes in ddY male mice: Possible role in protection against chemical the discovery of the antioxidant properties of naturally occurring carcinogenesis and toxicity. Pharmacology & Toxicology, 2003;92(1):33-38. phenolic compounds present therein. Curcumin exhibits strong 23. Jain SK. Ethnobotanical diversity in Zingibers of India., Ethnobotany, 1995;7(1– antioxidant activity6,17,22,41,42,48. 2):83–88. CONCLUSION 24. Jain JP, Bhathagar LS, Perishi MR, J. Res. India Med., Yoga Homoepathy. 1991;14:110p. Over all observation indicates the presence of free radical 25. Kapoor S and Priyadarsini KI. Protection of radiation-induced protein damage scavenging activity in all species (>50%), however Curcuma longa by curcumin. Biophys. Chem. 2001;92:119-126. and C.zedoaria are superior to the other three species. The activity 26. Kim KT, Yoo KM, Lee JW, Eom SH, Hwang IK, Lee CY. Protective effect of could be attributed to superior reducing power of both phenol and steamed American ginseng (Panax quinquefolius L.) on V79-4 cells induced by oxidative stress. J. Ethnopharm, 2007;111:443-445. curcumin content. Moderate antioxidant activity is possible due to 27. Kirtikar KR and Basu BD. Indian Medicinal Plants. vol. II. Internat. Book 95% oil contents having major constituents camphor, curzerenone, Distributors, Dehra, Dun, 1988. methyl eugenol, α-muurolene, pentadecanoic acid, (E,E)-farnesyl 28. Majeed M (Ed). Curcuminoids; antioxidant phytonutrients. NutriScience acetone and n-heptyl salicylate. The results indicate the higher Publishers, Inc., N.J., 1995. 29. Masuda T, Maekawa T, Hidaka K, Bando H, Takeda Y, and Yamaguchi H. medicinal use of C.longa and C.zedoaria due to its rich Chemical studies on antioxidant mechanism of curcumin. J. Agric. Food Chem., phytochemical contents. 2001;49 (5):2539-2547. ACKNOWLEDGMENT 30. Masuda T, Toi Y, Bando H, Maekawa T, Takeda Y, and Yamaguchi H. The authors are thankful to University Grants Commission New Structural identification of new curcumin dimmers and their contribution to the antioxidant mechanism of curcumin. J. Agric. Food Chem., 20;50(9):19-23. Delhi, for Financial Assistance in form of major research project to 31. Miquel J, Bernd A, Sempere JM, Diaz-Alperi J, and Ramirez A. The curcuma one of the author (R.K.S) we are also thankful to Head of the antioxidants: pharmacological effects and prospects for future clinical use. A Department of Botany and principal B.J.B. (A) College for review. Archives of Gerontology and Geriatrics, 2002;34:37-46. providing necessary facilities for carrying out the experimental 32. Nadkarni KM. Indian Medicinal Plants and Drugs- with their Medicinal Properties and Uses. Asiatic Publishing House New Delhi. 1998;450p. work. Finally we are thankful to Sabitri Nahak for helping in 33. Noguchi N, Komuro E, Niki E, and Willson RL. Action of curcumin as an computer work. antioxidant against lipid peroxidation. J. Japan oil Chem. Soc., REFERENCES 1994;43(12):1046-1051. 1. Aiyar YAK. Field crops in India. The Bangalore Printing and Publishing Co. 34. Okada K, Wangpoengtrakuk C, Tanaka T, Toyokuni S, Uchida K, and Osawa T. Ltd., Bangalore. 4thedition. 1954;319-326. Curcumin and especially tetrahydrocurcumin ameliorate oxidative stress induced 2. Ammon HPT and Wahl MA. Pharmacology of Curcuma longa. Planta Medica, renal injury in mice. J. Nutrition., 2001;131(8):2090-2095. 1991;57 (1):1-7. 35. Pothitirat W, Gritsanapan W. Variation of bioactive components in Curcuma longa in 3. Asai A and Miyazawa T. Dietary curcuminoids prevent high fat die–induced lipid Thailand.Curr.Sci., 2006;91(10):1397-1400. accumulation in rat liver and epididymal adipose tissue. J. Nutrition., 36. Purseglove JW. Tropical crops . Longman, London, 1968. 2001;131(11): 2932-2935. 37. Purseglove JW, Brown EG, Green CL and Robbin. Spices. Vol.II. Longman, 4. Asolkar LV, Kakkar KK and Chakre OJ. Second Supplement to Glossary of New York, Turmeric. 1981;532-580. Indian Medicinal Plants with Active Principles Part I (A-K). (1965-81). 38. Rao SM, Rao R. Flowering plants of Travancore, Vol. XIV. Government Press: Publications and Information Directorate (CSIR), New Delhi. 1992;414p. Trivendrum (reprinted by BSMP, Dehra Dun), 1914:400p. 5. Ammon HPT, Anazodo MI, Safayhi H, Dhawan BN, and Srimal RC. Curcumin: 39. Sadasivam S, Manickam A. Biochemical Methods 3/ed. New Age. 2007;284p. a potent inhibitor of leukotriene B4 formation in rat peritoneal 40. Sivarajan VV and Balachandran I. Ayurvedic drugs and their Plant Sources. polymorphonuclear neutrophils (PMNL). Planta Med., 1992;58:26-28. Oxford and IBH Publishing Co. Pvt. Ltd., New Delhi. 1994;570p. 6. Balasubramanyam M, Koteswari AA, Kumar RS, Monickaraj SF, Maheswari JU 41. Toda S, Miyase T, Arichi H, Tanizawa H, Takino Y. Natural antioxidants.III. and Mohan V. Curcumin-induced inhibition of cellular reactive oxygen species Antioxidative components isolated from rhizomes of Curcuma longa. Chem. generation: Novel therapeutic implications. J. Biosc., 2003;28:715-721. Pharm. Bull., 1985;33(4):1725-1728. 7. Bhandari SC, Vanaushadhi Chandrodaya. An Encyclopaedia of Indian Botanics 42. Sharma OP. Antioxidant activity of curcumin and related compounds. Biochem. and Herbs, Vol. Chaukhanabha Sanskrit Sansthan, Varanasi, 1992;7–8. Pharmacol., 1976;25:1811-1812. 8. Brand-Williams W. Cuvelier ME, and Berset C. Use of free radical method to 43. Song EK, Cho H, Kim JS, Kim NY, An NH, Kim JA, Lee SH, and Kim YCL. evaluate antioxidant activity. Lebensmittel Wissenschaft and Technologie, Diarylheptanoids with free radical scavenging and hepatoprotective activity in 1995;28(1):25-30. vitro from Curcuma longa. Planta-Medica, 2001;67(9):876-877. 9. Chavalittumrong P and Dechatiwongse T. Quality evaluation of turmeric. Thai. J.44. Sreejayan N, Rao MN, Curcuminoids as potent inhibitors of lipid peroxidation. J. Pharm. Sci., 1988;13:317-327 Pharm. Pharmocol., 1994;46:1013–1016. 10. Chavalittumrong P and Jirawattanapong W. Variation of active constituents of45. Srimal RC and Dhawan BN. Pharmacology of diferuloylmethane (curcumin), a Curcuma domestica rhizomes at different ages. Thai. J. Pharm. Sci., 1992;16:165-174. non-steroidal antiinflammatory agent. J. Pharm Pharmacol., 1973;25:447. 11. Chopra RN, Nayar SL, and Chopra IC. Glossary of Indian Medicinal Plants. 46. Thakur RS, Puri HS, and Husain A. Major Medicinal Plants of India, CIMAP, CSIR, New Delhi, 1980. Lucknow, India, 1989;50-52. 12. Duke JA. CRC Handbook of Medicinal Spices. CRC Press, Boca Raton, USA, 47. Ungphaiboon S, Supavita T, Singchangchai P, Sungkarak S, Rattanasuwan P, 2002 Tharat A. Study on antioxidant and antimicrobial activities of turmeric clear 13. Eigner D, Scholz D. Ferula asa-feotida and Curcuma longa in traditional liquid soap for wound treatment of HIV patients J.Sci. Technol., 2005;27(2):569- medicinal treatment and diet in . J. Ethnophamacol, 1999;67:1-6. 578. 14. Garg SK, Mathur VS, and Chudhury RR. Screening of Indian plants for anti- 48. Unnikrishnan MK and Rao MN. Inhibition of nitrite induced oxidation of fertility activity. Indian J. exp. Biol., 1978;16:1077. hemoglobin by curcuminoids. Pharmazie, 1995;50:490-492. 15. Gamble JS. Flora of the presidency of Madras. Vol.III. Bishen Singh Mahendra 49. Verghese J. Mango-ginger- an exotic flavourant. Indian Spices, 1990;27(2):15- Pal Singh, Dehra Dun, India. 1987:1478-1493. 16. 16. Nahak G and Sahu RK. Evaluation in comparative antioxidant activity of 50. Velayudhan KC, VK Muralidharan, VA Amalraj, RS Rana, B Singh and TA Curcuma longa & Curcuma aromatic. Natural Product: An Indian Jornal, Thomas. Genetic resources of Curcuma. NBPGR, Thrissur, 1994;74p. 2011;7(2):57-60. 51. Warrier PK, Nambiar VPK and Ramankutty C. Indian Medicinal Plants. Vol.1-5. 17. Govindarajan VS. Turmeric - Chemistry, technology, and quality. Crit. Rev. Orient Longman Ltd., Madras, 1993-1995. Food Sci. Nutr., 1980;12:199-301. 52. Watanabe S and Fukui T. Inhibitory effect of curcumin on the peroxisomes 18. Haines HH. Botany of Bihar and Orissa. 1961:1181p. proliferation by trichlro ethylene’. Japanese J. Toxicol. Environmental health, 19. Hirasa K and Takemasa M. Spice Science and Technology, Marcel Dekker Inc., 1997;43(1), 3. New York, 1998. 53. Watnabe S and Fukui T. Suppresive effect of curcumin on trichloroethylene- 20. Holtumn RE. The Zingiberaceae of Malay Peninsula. Gard. Bull.Singapore, induced oxidative stress’, J. Nutri. Sci. Vitaminol., 2000;46(5):230-234. 1950;13:1-249. 54. Watt, G.A. Dictionary of The Economic Products of India; Cosmo Publication, 21. Husain A, Virmani OP, Popli SP, Misra LN, Gupta MM, Srivastava GN, Delhi, India,1972; Vol. 6, p. 309. Abraham Z and Singh AK. Dictionary of Indian Medicinal Plants. CIMAP, 55. Nguyen TBT, Trinh DC, Do DR, Nguyen XD. Vietnam Hoa Hoc Va Cong Lucknow, India, 1992;546p. Nghiep Hoa Chat, 2001; 5:11–14 .

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011 Rajani Kanta Sahu et al. IRJP 2011, 2 (12), 243-248

56. Nguyen TBT, Trinh DC, Do DR, Nguyen XD. Vietnam Hoa Hoc Va Cong Nghiep Hoa Chat, 2001;4:28–32 . .

Table-1: Preliminary Phytochemical screening of Five Curcuma Species Phytoconstituents C.longa C.aromatica C.zedoaria C.angustifolia C.amada Alkaloids + + + + + Carbohydrates + + + + + Glycosides ------Terpinoid ------Protein & Amino acids + + + + + Fixed oils & fats ------Tannins ------Saponins + + + + + Steroids (Phytosterols) ------

Flavonoids + + + + + Phenols + + + + + Curcumin + + + + + Reducing sugar ------Anthraquinones + + + + + + = Denotes Present and -- = Denotes Absent

Table-2: Crude extracts, Phenol content, IC50Values, Curcumin content and Colour of Rhizomes of Curcuma Species C.longa C.aromatica C.zedoaria C.angustifolia C.amada Crude extracts(gm) 3.35 2.56 2.67 2.62 2.20

Phenol content(mg/ml) 285 205 195 110 50

IC50 Values(µg/ml) 24 120 40 100 121 Curcumin content 8.22 7.35 6.07 ------1.54 (mg/100mg) Colour of Rhizomes Yellow Creamy yellow Creamy White Creamy yellow yellow

Table-3: DPPH scavenging activity of Five Curcuma Species Conc. Antioxidant activity (%) of extracts(µg/ml) C.longa C.zedoaria C.angustifolia C.aromatica C.amada

40 67.69±0.04 52.11±0.07 45.52±0.03 44.61±0.06 47.69±0.04 80 69.23±0.05 54.45±0.04 48.61±0.02 47.61±0.03 49.23±0.05 120 70.76±0.04 58.00±0.06 52.00±0.08 50.00±0.07 50.76±0.04 160 72.30±0.03 61.07±0.01 56.07±0.07 53.07±0.02 51.30±0.03 200 74.61±0.02 63.27±0.05 58.35±0.04 55.38±0.06 52.61±0.02

Figure-1: Photograph showing morphology of different Curcuma Species

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011 Rajani Kanta Sahu et al. IRJP 2011, 2 (12), 243-248

Figure-2: DPPH scavenging activity of Five Curcuma Species in comparison to standard Ascorbic acid

Figure-3: Correlation between Phenol content, Curcumin content and Antioxidant activity of Five Curcuma Species

Source of support: Nil, Conflict of interest: None Declared

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011