S.K. Rodrigo,1 U.L.B. Jayasinghe2 AND B.M.R. Bandara1. 1Department of Chemistry, Faculty of Science, University of Peradeniya. 2Institute of Fundamental ...
Antifungal, Antioxidant and cytotoxic activity of ACRONYCHIA PEDUNCULATA and ADENANTHERA PAVONINA S.K. Rodrigo,1 U.L.B. Jayasinghe2 AND B.M.R. Bandara1 1
Department of Chemistry, Faculty of Science, University of Peradeniya 2 Institute of Fundamental Studies, Kandy
Introduction According to Ayurvedic Pharmacopoeia, Acronychia pedunculata (L.) Miq. (Rutaceae) and Adenanthera pavonina (L.) (Fabaceae) are widely used in various ayurvedic herbal preparations for treating diseases such as diarrhoea, tussis, asthma, ulcers, itchy skin, scales, sores, rheumatism, cold and cough (A. pedunculata) and boils, inflammations and gout (A. pavonina). Extracts of both plants exhibit antibacterial activity against methicillin-sensitive and –resistant strains of Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeruginosa (Jayasinghe et al., 2006). Methanol extracts of the stem and root bark exhibit significant cytotoxicity in the human KB tissue culture assay. Dichloromethane extract of stem-bark shows inhibition of cyclooxygenase-2 (COX-2) (Pathmasiri et al., 2005). In the present study the methanolic extracts of root, bark, leaves and fruits/seedkernels of both plants were examined for antifungal, antioxidant and cytotoxic activities.
Matale in March 2006, dried under mild sunlight inside the laboratory, and were separately extracted with methanol using soxhlet apparatus. Antifungal activity Antifungal activity of crude extracts was determined against Cladosporium cladosporioides by thin layer chromatography (TLC) autobiography method (Adikaram and Bandara, 1998). Antioxidant activity Antioxidant activity of crude extracts was determined using DPPH radical spectrophotometric assay using αtocopherol as the reference (Roberta et al., 2006). Antioxidant activity is expressed as the inhibition percentage and was calculated using the following formula: (1) % Inhibition = [(A0 -A t)/A0 ] x 100
(1)
where A0 = initial absorbance of DPPH and At = final absorbance of DPPH. Cytotoxicity
Materials and methods Preparation of extracts Root, bark, leaves and fruits/seedkernels of A. pedunculata (Ankenda) and A. pavonina (Madatiya) collected from locations close to Kandy and
Cytotoxicity of crude extracts was determined using the brine shrimp assay (Ajaiyeoba et al., 2006). LC50 values were determined using Probit Analysis.
Results Antifungal activity All methanolic extracts of A. pedunculata exhibited the presence of antifungal compounds against C. cladosporioides corresponding to the following Rf values on TLC bioassay with chloroform as the eluting solvent: 0.09 and 0.93 (root), 0.07 and 0.94 (leaf), 0.05, 0.22 and 0.96 (bark), 0.14 and 0.4 (fruit). Root, bark and seed extracts of A. pavonina exhibited the presence of antifungal compounds corresponding to the following Rf values: 0.22 and 0.90 (root), 0 (bark and seed-kernel). Antioxidant activity Percentage antioxidant activity of methanolic extracts of A. pedunculata and A. pavonina is given in Table 1. Cytotoxicity Cytotoxicity of each extract is given in Table 2. Discussion Extracts of A. pedunculata displayed strong antifungal properties and contained at least 6 antifungal compounds, while those of A. pavonina were marginally active; inhibition areas in the TLC-bioassay plate of A. pedunculata were sharper and much larger than those of A. pavonina. Stem-bark of A. pavonina showed significant antioxidant activity compared to that of the reference, α-tocopherol while root, stem-bark and fruit of A. pedunculata and root of A. pavonina showed moderate activity. Stem-bark and fruit extracts of A. pedunculata showed significant cytotoxicity against brine
shrimp Artemia salina. Only the root and stem-bark extracts of A. pavonina showed cytotoxicity. Conclusion The results show that Acronychia pedunculata contains strongly active antifungal compounds and cytotoxic principles. The bark of Adenanthera pavonina contains antioxidant compound(s) presumably more active than α-tocopherol. The results of this and previous studies corroborate with the use of these plants in traditional medicine. References Adikaram, N.K.B. Bandara, B.M.R., 1998. Methodology for studying defence mechanism against fungal pathogens: an overview. ACIAR proceedings, Canberra, 80:181184. Ajaiyeoba, E.O. Abiodium, O.O. Falade, M.O. Ogbole, N.O. Ashidi, J.S. Happi, C.T. Akinboye, D.O., 2006. Invitro cytotoxicity studies of 20 plants used in Nigerian antimalarial ethnomedicine. Phytomedicine, 13:295-298. Jayasinghe, P.K.I.D.E. Bandara, B.M.R. Ekanayaka, E.W.M.A. Thevanesam, V., 2006. Screening for Antimicrobial activity of Acronychia pedunculata (Ankenda) and Adenanthera pavonina (Madatiya) against bacteria causing skin and wound infections in humans. Proceedings of the Peradeniya University Research Sessions, Sri Lanka, 11:105. Olumayokun, A. O.Echianu, C.A.. A.E. Aduragbemi, A. A. Makinde, J. M., 2004. Studies on Adenanthera pavonina seed extract. Inflammopharmacology, 12:197-202. Pathmasiri, W., El-Seedi, H.R. Han, X. Janson, J. Huss, U. Bohlin, L., 2005. Aryl ketones from
Acronychia pedunculata with Cyclooxygenase-2 Inhibitory Effects. Chemistry & Biodiversity, 2:463-469. Roberta, R. Luciana, G.M. Luciana, C.C. Gláucia, P., 2006. Evaluation of the antioxidant Properties of the Brazilian Cerrado Fruit Annona
crassiflora (Araticum). Journal of Food Science, 71:102-107. We thank Mr. W. Pathmasiri, Uppsala University, Sweden for providing valuable literature material.
Table 1. Percentage antioxidant activity of crude extracts of Acronychia pedunculata and Adenanthera pavonina in DPPH assay
A. pedunculata A. pavonina#
Root
Leaves
Stem-bark
Fruits/Seed-kernels#
14.9±0.3 31.2±0.4
8.0±0.6 5.9±1.8
19.8±1.2 70.3±0.5
17.1±1.1 2.1±0.1
Table 2. Cytotoxicity of Acronychia pedunculata and Adenanthera pavonina extracts given as percentage mortality against brine shrimp Artemia salina
Plant
Extract
A. pedunculata
A. pavonina #
Root Stem-bark Leaves Fruits Root Stem-bark Leaves Seed-kernels
Concentration of extracts/ppm 1000 500 250 100 50 100 87 67 30 0 100 100 100 87 53 80 47 0 0 0 100 100 100 87 57 100 100 87 57 0 100 77 37 0 0 0 0 0 0 0 0 0 0 0 0
#
25 0 0 0 0 0 0 0 0
LC50/ ppm 174 54 587 51 108 304 0 0
As determined by Probit Analysis, using EPA Probit Analysis Program, Version 1.5
