A Comparative Phytochemical and Physicochemical Evaluation: Tikta-anguna, Kiri-anguna and Jeewanti Sujatha Hewageegana Menuka Arawwawala Anurakumara Tissera Sami Ariyawansa Induragare Dammaratana

Introduction Objectives Materials and Methods Results Conclusion

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

Wattakaka volubilis Tikta anguna Kiri anguna (Behet anguna)

Leaves, roots and tender stalks are used 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

Jeewanti Leptadenia reticulate (Retz) Wight and Arn. Family : Asclepiadaceae A much - branched twining shrub, branches numerous and younger ones glabrous, Leaves thin, ovate

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

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

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

Comparison of the Two plants L. reticulate and W. volubilis Family Asclepiadaceae Classical names Jivanti, Svarnajivanti, Tittajivanti Hemajivanti, Sujivanti, Svarnajivantika Morphology Many similarities Common uses Skin ailments Respiratory tract infections ENT disorders Lactogenic Burning sensation Antipyretic Use for poisoning Aphrodisiac Increase urination

Purpose of this study was to investigate the possibility of using Tikta-anguna and Kiri-anguna as substitutes for Jeewanti

Objectives To compare the phytochemical parameters of W. volubilis (Tikta-anguna and Kiri-anguna) and L. reticulata (Jeewanti) To compare the physicochemical parameters of To compare the fingerprint profiles of W. volubilis (Tikta-anguna and Kiri-anguna) and L. reticulata (Jeewanti)

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

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  

Screening of Phytochemical compounds Presence /Absence of Phenolic compounds Flavanoids Tannins Coumarins Alkaloids Saponins Steroid glycosides

Physicochemical parameters were evaluated according to WHO standards Determination of physicochemical parameters Physicochemical parameters were evaluated according to WHO standards leMatter

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

1. % Total ash = Ash Weight x 100 Ash Content : 1. % Total ash = Ash Weight x 100 Weight of sample 2. % Acid-insoluble ash = Acid insoluble Ash Weight x 100 3. % Water-soluble ash = Total Ash Weight - water insoluble residue in total ash x 100

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

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

Presence or Absence of Phytochemicals Classes Results Table 1 : Phytochemical classes of W. volubilis varities and L. reticulata Phytochemical Classes Presence or Absence of Phytochemicals Classes Tikta-anguna Kiri-anguna (Behet-anguna) Jeewanti (HWE) (HEE) (CWE) (CEE) Phenolic compounds  Saponins - Flavanoids Steroid glycosides Tannins Coumarin Alkaloids

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

Physico-chemical parameters W. Volubilis (%) - (Dry wt basis) Table 2: Physicochemical parameters of Wattakaka volubilis and Leptadenia reticulata Physico-chemical parameters W. Volubilis (%) - (Dry wt basis) L. Reticulata (%) (Dry wt basis) (Jeewanti) Tikta anguna Kiri anguna (Behet anguna) 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

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

3. TLC fingerprint profiles of Tikta-anguna, Kiri-anguna and Jeewanti 1 2 3 2µL 2µL 2µL 1 2 3 2µL 2µL 2µL Wave length – at 254 nm After spraying Vanillin -sulphuric acid L. reticulata (Jeewanti ) 2. W. volubilis (Kiri-anguna) 3. W. volubilis (Tikta-anguna)

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)

According to TLC fingerprints of Tikta-anguna, Kiri-anguna and Jeewanti, similar compounds were identified in three samples

According to the results of this study, phytochemical parameters, physicochemical parameters and fingerprint profiles of Tikta-anguna and Kiri-anguna have very similar properties and they are similar to Jeewanti also

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

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 (behet-anguna) is much better to use as a substitute for Jeewanti

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

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)

Thank you

Jeewanti Tikta-anguna Kiri-anguna (Bitter verity) (Sweet verity) Rasa Madhura Katu, Tikta Madhura Guna Laghu, Snigdha Ruksha, Laghu Laghu, …… Veerya sheeta Ushna Sheeta Vipaka Madhura Katu Madhura Prabhawa Jivniya