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Pharmacognostical Studies on the Root and Rhizome of Nymphoides Hydrophylla (Linn.) O

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Pharmacognostical Studies on the Root and Rhizome of Nymphoides Hydrophylla (Linn.) O Indian Journal of Natural Products and Resources Vol. 3(3), September 2012, pp. 371-385 Pharmacognostical studies on the root and rhizome of Nymphoides hydrophylla (Linn.) O. Kuntze –An alternate source for Tagara drug V Madhavan1, M Jayashree1, S N Yoganarasimhan1*, M Gurudeva2, R Deveswaran3 and R Mythreyi1 1Department of Pharmacognosy, MS Ramaiah College of Pharmacy, Bangalore-560 054, India 2Department of Botany, VV Pura College of Science, Bangalore 3Department of Pharmaceutics, MS Ramaiah College of Pharmacy, Bangalore Received 13 October 2011; Accepted 19 January 2012 Tagara is an important drug used in Ayurvedic medicine for the treatment of several diseases. The accepted botanical source of Tagara is Valeriana jatamasni Jones, although different species of Nymphoides Hill are used by the physicians. The pharmacognostical evaluation of the root and rhizome of Nymphoides hydrophylla, a potential alternative source for Tagara is presented in this paper. Important details like morphology of the plant, macro-, microscopical characters, macerate, histochemical tests, UV studies of the root and rhizome along with physico-chemical constants, phytochemical analysis and HPTLC finger print profile are presented, all of which will be useful in the standardization of this drug. Isolation of β-sitosterol, betulinic, salicylic and tannic acids are reported for the first time from N. hydrophylla. The pharmacognostical and phytochemical studies help in the identification of N. hydrophylla from other species used as Tagara. Keywords: Nymphoides hydrophylla, Pharmacognosy, Root, Rhizome, HPTLC, Tagara. IPC code; Int. cl. (2011.01)—A61K 36/00 Introduction (Lour.) O. Kuntze, N. indica (Linn.) O. Kuntze and Tagara is an important drug used in Ayurvedic N. macrosperumum Vasudevan3. N. indica is used as medicine for the treatment of diseases like anaemia an antiscorbutic and febrifuge, and as a substitute for (pandu), epilepsy (apasmara), fever (jwara), jaundice chiretta [Swertia chirayita (Roxb. ex Flem.) Kars.] to (kamala), mental disorders (unmada), tuberculosis treat fever and jaundice4. Literature review revealed (yakshma) and also as a general and brain tonic1. that while pharmacognostical studies have been carried Valeriana jatamansi Jones (Valerianaceae) is the out on the roots and rhizome of N. macrospermum5 and accepted botanical source of Tagara which constitutes N. indica6, no work is available on N. hydrophylla7-9. as one of the ingredients in ayurvedic formulations like Hence, the present investigation on the root and Bala taila, Bilvadi gutika, Dasanga lepa, Jatiphaladya rhizome of N. hydrophylla was undertaken. curna, Karpuradyarka, Maha kalyanaka ghrita, Maha Materials and Methods narayana taila, Puga khanda, Sahacaradi taila, 2 Plant material was collected from a water tank in the Srikhandasava, to name a few . In South India, a drug vicinity of Nagercoil, Kanyakumari district, Tamil under the name Granthika tagara (Kannada), Nadu, during November 2007, processed into botanically identified as Nymphoides macrospermum herbarium specimen10; voucher herbarium specimen Vasudevan (Menyanthaceae) is used in place of (Jayashree 025) and a sample of the crude drug are Tagara (Valeriana jatamansi) for the same ayurvedic preserved at the herbarium and crude drug museum of preparations, under the same formulations supporting the Department of Pharmacognosy, M S Ramaiah the fact that Granthika tagara (N. macrospermum) may College of Pharmacy, Bangalore (MSRCP). The plant have similar therapeutic properties to that of Tagara material was identified following local floras11-13 and (V. jatamansi); furthermore, it is often found that authenticated by S N Yoganarasimhan, Plant Granthika tagara is an admixture of different species Taxonomist. For pharmcognostical studies, a small of Nymphoides Hill found in S. India, viz. quantity of rhizome and root were preserved in 70% N. aurantiacum (Dalz.) O. Kuntze, N. hydrophylla alcohol; free hand sections were taken for macro- and ___________ microscopical observations14,15. Photomicrographs *Correspondent author: E-mail: [email protected]; [email protected] were obtained by observing the free hand sections Phone: +91-80-23608942 under a compound binocular microscope (Olympus- 372 INDIAN J NAT PROD RESOUR, SEPTEMBER 2012 CH20i model) with a built-in analogue camera Gen. and Spec. Gent. 342. 1839; Clarke in Hook. f., Fl. (CMOF, 1.4 mega pixel). Computer Images were Brit. India, 4: 131. 1883; Gamble, Fl. Pres. Madras, 2: captured using AV Digitaliser fitted with a Grand VCD 883, 2005 (reprint); Menyanthes hydrophylla Lour., Fl. 2000–Capture Guard. Measurements of cells and Cochinch. 1:129, 1790. tissues (in µm) of the transverse section and macerate Aquatic herbs with long floating stem (stolon). were carried out using Micro Image Lite Image Rhizome short, erect, with petiole-like branches Analysis Software (Cybernetics, Maryland, USA) and reaching the surface of water and producing a node are provided in parenthesis. Physicochemical studies, from which arise a tuft of adventitious roots, a cluster ultra-violet and chromatographic studies were carried of flowers, a single floating leaf and a single branch, out following standard methods16-20. Vernacular names which again proceeds similarly. Leaves orbicular, are provided following Gurudeva21. Isolation of deeply cordate at base, often purplish, to 10 cm in β-sitosterol was carried out as per Gerhardt et al22. The diameter, lobes rounded, faintly crenate, pale green total phenolic compounds were determined following above, glandless and with prominent nerves beneath; Folin-Ciocalteace method23 and HPTLC was carried petioles to 3.5 cm long. Flowers white, with yellow out using a Camag HPTLC system with Linomat V centre, delicate, with a ring of white hairs round the sample applicator, a Camag 3 TLC Scanner and throat, numerous, in dense clusters; pedicels to 6 cm WINCATS 4 software for interpretation of data. An long, unequal. Calyx divided almost to the base, aluminum plate (20 × 10 cm) pre-coated with silica gel oblong-lanceolate, obtuse. Corolla white, when 60F254 (E Merck) was used as adsorbent. The plates expanded; lobes obovate, rounded at apex, glabrous, were developed using toluene:chloroform:ethanol with a broad longitudinal crest down the middle of (28.5:57:14.5) for alcohol and aqueous extracts in a each lobe, margins not ciliate. Capsule broadly ovoid Camag twin trough chamber and scanned under 254, or subglobose. Seeds numerous with prominent, small, 366 and 425 nm. The developed plate was derivatised slightly glochidiate tubercles, scabrous, pale yellowish- by dipping in Libermann Burchard’s reagent and brown (Plates 1, 2). drying at 110°C in a hot air oven for 10 min, scanned Nymphoides hydrophylla usually flowers and fruits in under all 3 wavelengths and photo documented both the winter season. In this species polystely occurs in the before and after derivatisation using a Camag 3 cortical region of the stolon and in the petiole of the Reprostar. For the identification of phenolic leaves. It is very common throughout India and is compounds, 10 µl of tannic acid solution and 5 µl of gregarious in habit, generally growing near the margins salicylic acid solution were applied and developed with of tanks and lakes. Also found in Sri Lanka, China and ethyl acetate: benzene: formic acid (9:11:0.5) and Malaysia26. scanned under 254, 366 and 425 nm. The developed Vernacular names plate was photo documented at 254 nm. For the Tamil: Akacavalli; Sanskrit: Kumudvathi, Seethali, identification of betulinic acid, both extracts were Kalanu saariva, Tagara; Kannada: Anthara thaavare; developed with n-hexane: ethyl acetate: glacial acetic Marathi: Kolare chikal; Hindi: Tagarmul, Cumuda, acid (7:3:0.03), derivatised with Anisaldehyde Ghainchu, Kumudini, Lodh, Baank; Telugu: Anthara (Ref.24) H2SO4 , scanned at 425 nm and photodocumented. thaarama, Chiri alli, Pitta kaluva; Bengali: Panchuli, For the identification of β-sitosterol, toluene:diethyl Chandmalla; Malayalam: Tsjeroea–citamber; ether (40:40) was used, derivatised with Vanillin- Manipuri: Tharo–macha. (Ref. 25) H SO dried at 105°C , scanned at 425 nm and 2 4, Macro- and microscopical characters of the root and rhizome photo documented. The Rf values and peak area of the Macroscopical characters of the root extracts and the isolated sample were interpreted by Roots arise in tufts all around the rhizome, running using a WINCATS 4 software. The developed plates vertically downwards, very slender, spongy, brownish, were photo documented under 425 nm using a Camag measuring to 32 cm long when fresh, and has no 3 Reprostar. All the solvents used were of HPLC grade. characteristic odour or taste. Young fresh roots are minutely hairy and white to brownish while dry roots Results are slender, brownish (Plates 3, 4). Plant morphology Nymphoides hydrophylla (Linn.) O. Kuntze, Rev. Macroscopical characters of the rhizome Gen. Pl. 429. 1891. Menyanthes cristata Roxb. Pl. Cor. The rhizomes are slender, running horizontally, 2: 3. t. 105, 1798; Limnanthemum cristatum Griseb. cone-like with many leaf bases, rough, brittle, brownish MADHAVAN et al: PHARMACOGNOSY OF NYMPHOIDES HYDROPHYLLA 373 without, white within, with several fragments of roots Microscopical characters of the root arising from different points, measuring to 9 cm long; A transverse section of the root is wheel-like and it possesses no characteristic odour or taste; dry exhibits an epidermis, an outer, middle and inner rhizomes are blackish
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