A Comparative Phytochemical and Physicochemical Evaluation: Tikta-anguna, Kiri-anguna and Jeewanti Sujatha Hewageegana Menuka Arawwawala Anurakumara Tissera Sami Ariyawansa Induragare Dammaratana 1 Introduction Objectives Materials and Methods Results Conclusion 2 Introduction Tikta-anguna and Kiri-anguna (Behet anguna) Both come under one botanical name Wattakaka volubilis (Linn.f) Family: Asclepiadaceae • Both creepers are large twining shrubs, with long glabrous branches, Leaves thin, ovate 1. Bitter variety 2. Sweet variety 3 Wattakaka volubilis Tikta anguna Kiri anguna (Behet anguna) 4 • Parts Used: • Leaves, roots and tender stalks are used • Actions and common uses: • Leaves are used for boils and abscesses as an application • Roots and tender stalks are used for expectorant properties in Ayurveda and traditional medicine • Lactogenic • Aphrodisiac • Sweet variety –cooling and Tonic 5 Jeewanti Leptadenia reticulate (Retz) Wight and Arn. Family : Asclepiadaceae A much - branched twining shrub, branches numerous and younger ones glabrous, Leaves thin, ovate 6 L. reticulate • Parts Used: Both roots and leaves Actions and common uses: Both roots and leaves - Skin infections, wounds Leaves - cough, as a respiratory stimulant, Lactogenic Aphrodisiac Cooling and Tonic 7 Research background : Ayurvedic texts contain lot of simple and effective as well as both externally and internally used medicinal preparations, containing Jeewanti (L. reticulata) Jeewanti is found in most parts of India, but it is not available in Sri Lanka We had to find a most suitable substitute for Jeewanti 8 • According to literature survey and the conversation with Senior Scientist in charge, Haldummulla Research Gardens, we were able to find morphologically, nearly similar two plants (Tikta-anguna and Kiri-anguna) for Jeewanti • In addition, they have similar practices for disease conditions 9 Comparison of the Two plants L. reticulate and W. volubilis L. reticulate W. volubilis Family Asclepiadaceae Asclepiadaceae Classical names Jivanti, Svarnajivanti, Tittajivanti Hemajivanti, Sujivanti, Svarnajivantika Morphology Many similarities Many similarities Common uses Skin ailments Respiratory tract infections ENT disorders Lactogenic Burning sensation Antipyretic Use for poisoning Aphrodisiac Increase urination Skin ailments Respiratory tract infections ENT disorders Lactogenic Burning sensation Antipyretic Use for poisoning Aphrodisiac Increase urination 10 Purpose of this study was to investigate the possibility of using Tikta-anguna and Kirianguna as substitutes for Jeewanti 11 Objectives To compare the phytochemical parameters of W. volubilis (Tikta-anguna and Kiri-anguna) and L. reticulata (Jeewanti) To compare the physicochemical parameters of W. volubilis (Tikta-anguna and Kiri-anguna) and L. reticulata (Jeewanti) To compare the fingerprint profiles of W. volubilis (Tikta-anguna and Kiri-anguna) and L. reticulata (Jeewanti) 12 Materials and Methods W. volubilis (both varieties) were collected from Colombo, Sri Lanka Authentication was done by Scientist , Botany Division, BMARI, Navinna, Maharagama Voucher specimen No : W 2561 L. reticulata was collected from Jamnagar, India Authentication was done by Botanist, Vidyaratnam Foundation (Research and Development Section), Thrissur, Kerala, India, according to the standards of Ayurveda Pharmacopeia in India 13 Plant materials • Plant materials (leaves with tender stalk) were shade dried and cut into small pieces Then prepared the powder using the grinder • Materials were kept in air tight containers until used 14 Screening of Phytochemical compounds Presence /Absence of • Phenolic compounds Flavanoids Tannins • Coumarins • Alkaloids • Saponins • Steroid glycosides 15 • Determination of physicochemical parameters Physicochemical parameters were evaluated according to WHO standards 16 Extractable matter in Hot Water Extract (HWE) Hot Ethanol Extract (HEE) Cold Water Extract (CWE) Cold Ethanol Extract (CEE) % Extractable mater = Weight of extract x 100 Weight of sample 17 Ash Content : 1. % Total ash = Ash Weight x 100 Weight of sample 2. % Acid-insoluble ash = Acid insoluble Ash Weight x 100 Weight of sample 3. % Water-soluble ash = Total Ash Weight - water insoluble residue in total ash x 100 Weight of sample 18 TLC fingerprint profiles of W. volubilis and L. reticulata • Extract - methanol extract • Plant material - 4.0 g • Extracts were spotted on a silica gel plate and TLC fingerprints were developed using Dichloromethane: Ethyl acetate: Cyclohexane (40:1:10 v/v/v) • Vanillin-sulphuric acid reagent was used for colour development 19 Statistical analysis • Data were analyzed by using Mann Whitney test and findings of p<0.05 was considered to indicate statistical significance • All data were presented as mean ± SEM • All the values were express as dry weight of the sample and they were performed in triplicate for accuracy 20 Results Table 1 : Phytochemical classes of W. volubilis varities and L. reticulata Phytochemical Presence or Absence of Phytochemicals Classes Tikta-anguna Classes Kiri-anguna (Behet-anguna) Jeewanti (HWE) (HEE) (CWE) (CEE) (HWE) (HEE) (CWE) (CEE) (HWE) (HEE) (CWE) (CEE) Saponins - - - - - - Flavanoids - - - - - - - - - - Steroid glycosides Tannins Coumarin - - - - Alkaloids - - - - - - - - Phenolic compounds 21 - • Screening of phytochemical compounds of (a) cold and hot ethanol and water extracts of W. volubilis and L. reticulata revealed the presence of phenolic compounds, tannins, and steroids (b) cold and hot water extracts of W. volubilis and L. reticulata revealed the presence of saponins (c) cold and hot ethanol and water extracts of L. reticulata and hot extracts of W. volubilis revealed the presence of coumarin (d) In addition, hot water extract of W. volubilis contained flavanoids 22 Table 2: Physicochemical parameters of Wattakaka volubilis and Leptadenia reticulata Physico-chemical W. Volubilis (%) - (Dry wt basis) L. Reticulata (%) parameters Tikta anguna Kiri anguna (Behet anguna) (Dry wt basis) (Jeewanti) Hot water extractable matter 37.37± 0.56 31.67 ± 0.33 31.54 ± 0.46 Hot ethanol extractable matter 19.62± 0.10 3.36 ± 0.05 13.19 ±0.42* Cold water extractable matter 7.99± 0.04 7.01± 0.28 5.99 ± 0.49 Cold ethanol extractable matter 1.55± 0.02 1.37 ± 0.05 6.34± 0.13* Total ash content 11.80± 0.07 16.46± 0.11 16.63 ± 0.30* Acid-insoluble ash Content 0.91± 0.02 0.81± 0.01 2.30 ± 0.02* Water-soluble ash Content 5.76± 0.21 6.11 ± 0.06 6.13 ± 0.03 23 Following three physicochemical parameters of Jeewanti were significantly different from that of Tikta-anguna and Kiri-anguna 1. Hot and Cold Ethanol Extractable matter 2. Total ash Content 3. Acid-insoluble ash Content 24 3. TLC fingerprint profiles of Tikta-anguna, Kiri-anguna and Jeewanti 1 2 3 1 2 2µL 2µL 2µL 2µL 2µL Wave length – at 254 nm 3 2µL After spraying Vanillin -sulphuric acid 1. L. reticulata (Jeewanti ) 2. W. volubilis (Kiri-anguna) 3. W. volubilis (Tikta-anguna) 25 TLC- densitogram fingerprint of Tikta-anguna, Kiri-anguna and Jeewanti, at 254 nm Red – Tikta anguna (W. volubilis) Blue – Kiri anguna (W. volubilis) Green – Jeewanti (L. reticulata) 26 According to TLC fingerprints of Tikta-anguna, Jeewanti, Kiri-anguna similar and compounds were identified in three samples 27 According to the results of this study, phytochemical parameters, physicochemical parameters and fingerprint profiles of Tiktaanguna and Kiri-anguna have very similar properties and they are similar to Jeewanti also 28 conclusions • As this study reveals, Tikta-anguna, Kiri-anguna and Jeewanti have more similarities in terms of phytochemical parameters, physicochemical parameters and fingerprint profiles • Therefore, both Tikta-anguna and Kiri-anguna can be used as substitutes for Jeewanti in ayurvedic preparations 29 According to traditional medicine, Kiri-anguna (Behet-anguna) is commonly used for medicinal preparations Further, Kiri-anguna (behet-anguna) can be considered as the “sweet variety” of W. volubilis and it has similar properties such as GUNA (Quality), RASA (Taste), VIPAK (Metabolism), VIRYA (Potency), PRABHAV (Impact) to Jeewanti • After considering all, we can conclude that kiri-anguna (behetanguna) is much better to use as a substitute for Jeewanti 30 References • Parangepe, P., Indian medicinal Plants, 2005, Chaukhamba Sanskrit Pratishthan, Delhi.pp 116-117 • Jayaweera, D.M.A., Medicinal Plants (Indigenous and Exotic) Used in Ceylon, 2006, The National Science Foundation, Sri Lanka, pp 225 • Kirtikar, K. R., Basu, B. D., Indian Medicinal Plants- volume III, 1996,Valley offset printers and publishers, Dehra Dun, pp 2267-2268 • Senaratna, L. K., A Check List of Flowering Plants of Sri Lanka, 2001, The National Science Foundation, Sri Lanka, pp 46 • Sharma, P.C., Yelne, M.B., Dennis, T.J., Database on Medicinal plants used in Ayurveda, 2002, Central Council for Research in Ayurveda and Siddha, New Delhi. Pp 270-272 31 Acknowledgement • Mr Piyal Marasingha – Senior scientist , Dept of Ayurveda • Dr. Sudeesh Kumar and the Vidyaratnam Foundation (Research and Development Section), Thrissur, Kerala, India • Industrial Technology Institute • National Centre for Advanced Studies for Humanities and Social Sciences (NCAS) 32 33 • • • • • Rasa Guna Veerya Vipaka Prabhawa » Jeewanti Tikta-anguna (Bitter verity) Kiri-anguna (Sweet verity) Madhura Laghu, Snigdha sheeta Madhura Jivniya Katu, Tikta Ruksha, Laghu Ushna Katu Madhura Laghu, …… Sheeta Madhura