Ruta graveolens L. is a medicinal plant used in traditional systems of medicine for treatment of psoriasis, vitiligo, leucoderma, and lymphomas with well-known anti-inflammatory and anticancer properties. Therefore antioxidant potential of R. graveolens (in planta and in vitro) was investigated. As antioxidants present in plant extracts are multifunctional, their activity and mechanism depends on the composition and conditions of the test system. Therefore, the total antioxidant capacity was evaluated using assays that detect different antioxidants: free radical scavenging (DPPH and ABTS), transition metal ion reduction (phosphomolybdenum assay), reducing power, and nitric oxide reduction. Content of furanocoumarin-bergapten in the extracts showed good corelation with free radical scavenging, transition metal reduction and reducing power, while total phenolic content showed good corelation with nitric oxide reduction potential. Antioxidant activity of in vitro cultures was significantly higher compared to in vivo plant material. The present study is the first report on comprehensive study of antioxidant activity of R. graveolens and its in vitro cultures. 1. Introduction Free radicals, together with secondarily formed radicals, are known to play an important role in the pathogenesis of many chronic conditions like atherosclerosis, arthritis, diabetes, ischemia, reperfusion injuries, central nervous system injury, and cancer [1, 2]. Hence, the study of antioxidant status during a free radical challenge can be used as an index of protection against the development of these degenerative processes in experimental condition for therapeutic measures. Ruta graveolens is used in homeopathic, ayurvedic, and unani preparations [3] because this herb is so efficacious in various diseases (Ruta derived from Greek “reuo” means to set free). It has been extensively used in treatment of leucoderma, vitiligo, psoriasis, multiple sclerosis, cutaneous lymphomas, rheumatic arthritis and recently reported to possess anti-inflammatory and anticancer activity [4, 5]. Antioxidants in plants are affected by area, climatic conditions, and pest attack [6, 7]; therefore in vitro cultures are being investigated as alternate source of natural antioxidants [8]. For estimation of total antioxidant potential many authors have stressed the need to perform more than one type of antioxidant activity measurement to take into account the various mechanisms of antioxidant action [9]. With this perspective the present study investigates the total antioxidant activity evaluated using DPPH,
References
[1]
N. C. Cook and S. Samman, “Flavonoids—Chemistry, metabolism, cardioprotective effects, and dietary sources,” Journal of Nutritional Biochemistry, vol. 7, no. 2, pp. 66–76, 1996.
[2]
J. T. Kumpulainen and J. T. Salonen, Natural Antioxidants and Anticarcinogens in Nutrition, Health and Disease, Royal Society of Chemistry, Cambridge, UK, 1999.
[3]
K. C. Preethi, C. K. K. Nair, and R. Kuttan, “Clastogenic potential of Ruta graveolens extract and a homeopathic preparation in mouse bone marrow cells,” Asian Pacific Journal of Cancer Prevention, vol. 9, no. 4, pp. 763–769, 2008.
[4]
S. Pathak, A. Multani, P. Banerji, and P. Banerji, “Ruta 6 selectively induces cell death in brain cancer cells but proliferation in normal peripheral blood lymphocytes: a novel treatment for human brain cancer,” International Journal of Oncology, vol. 23, no. 4, pp. 975–982, 2003.
[5]
K. Preethi, G. Kuttan, and R. Kuttan, “Anti-tumour activity of Ruta graveolens extract,” Asian Pacific Journal of Cancer Prevention, vol. 7, no. 3, pp. 439–443, 2006.
[6]
G. Miliauskas, P. R. Venskutonis, and T. A. van Beek, “Screening of radical scavenging activity of some medicinal and aromatic plant extracts,” Food Chemistry, vol. 85, no. 2, pp. 231–237, 2004.
[7]
X. Liu, M. Zhao, J. Wang, B. Yang, and Y. Jiang, “Antioxidant activity of methanolic extract of emblica fruit (Phyllanthus emblica L.) from six regions in China,” Journal of Food Composition and Analysis, vol. 21, no. 3, pp. 219–228, 2008.
[8]
M. Gulluce, M. Sokmen, D. Daferera et al., “In vitro antibacterial, antifungal, and antioxidant activities of the essential oil and methanol extracts of herbal parts and callus cultures of Satureja hortensis L,” Journal of Agricultural and Food Chemistry, vol. 51, no. 14, pp. 3958–3965, 2003.
[9]
E. N. Frankel and A. S. Meyer, “The problems of using one-dimensional methods to evaluate multifunctional food and biological antioxidants,” Journal of the Science of Food and Agriculture, vol. 80, no. 13, pp. 1925–1941, 2000.
[10]
R. Diwan and N. Malpathak, “Histochemical localization in Ruta graveolens cell cultures: elucidating the relationship between cellular differentiation and furanocoumarin production,” In Vitro Cellular and Developmental Biology, vol. 46, no. 1, pp. 108–116, 2010.
[11]
R. Diwan and N. Malpathak, “Novel technique for scaling up of micropropagated Ruta graveolens shoots using liquid culture systems: a step towards commercialization,” New Biotechnology, vol. 25, no. 1, pp. 85–91, 2008.
[12]
V. Singleton, R. Orthofer, and R. Lamuela-Raventos, “Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent,” in Methods in Enzymology, L. Packer, Ed., vol. 299, pp. 152–315, Academic Press, San Diego, Calif, USA, 1999.
[13]
M. R. Szabo, C. Idi？oiu, D. Chambre, and A. X. Lupea, “Improved DPPH determination for antioxidant activity spectrophotometric assay,” Chemical Papers, vol. 61, no. 3, pp. 214–216, 2007.
[14]
C. Teow, V. Truong, R. McFeeters, R. Thompson, K. Pecota, and C. Yencho, “Antioxidant activities, phenolic and β-carotene contents of sweet potato genotypes with varying flesh colours,” Food Chemistry, vol. 103, no. 3, pp. 829–838, 2007.
[15]
P. Prieto, M. Pineda, and M. Aguilar, “Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E,” Analytical Biochemistry, vol. 269, no. 2, pp. 337–341, 1999.
[16]
M. Oyaizu, “Studies on products of the browning reaction. Antioxidative activities of browning reaction products prepared from glucosamine,” Japanese Journal of Nutrition, vol. 44, no. 6, pp. 307–315, 1986.
[17]
L. Marcocci, J. Maguire, M. Droy-Lefaix, and L. Packer, “The nitric oxide-scavenging properties of Ginkgo biloba extract EGb 761,” Biochemical and Biophysical Research Communications, vol. 201, no. 2, pp. 748–755, 1994.
[18]
A. Karagozler, B. Erdag, Y. Emek, and D. Uygun, “Antioxidant activity and proline content of leaf extracts from Dorystoechas hastate,” Food Chemistry, vol. 111, pp. 400–447, 2008.
[19]
X. Piao, I. Park, S. Baek, H. Kim, M. Park, and J. Park, “Antioxidative activity of furanocoumarins isolated from Angelicae dahuricae,” Journal of Ethnopharmacology, vol. 93, no. 2, pp. 243–246, 2004.
[20]
I. Grzegorczyk, A. Matkowski, and H. Wysokinska, “Antioxidant activity of extracts from in vitro cultures of Salvia officinalis L,” Food Chemistry, vol. 104, no. 2, pp. 536–541, 2007.
[21]
F. Borges, F. Roleira, N. Milhazes, L. Santana, and E. Uriarte, “Simple coumarins and analogues in medicinal chemistry: occurrence, synthesis and biological activity,” Current Medicinal Chemistry, vol. 12, no. 8, pp. 887–916, 2005.
[22]
I. Kostova, “Synthetic and natural coumarins as cytotoxic agents,” Current Medicinal Chemistry, vol. 5, no. 1, pp. 29–46, 2005.
[23]
R. Marwah, M. Fatope, R. Al Mahrooqi, G. B. Varma, H. Al Abadi, and S. Al-Burtamani, “Antioxidant capacity of some edible and wound healing plants in Oman,” Food Chemistry, vol. 101, no. 2, pp. 465–470, 2007.
