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Brine shrimp lethality bioassay of selected Indian medicinal plants
Fitoterapia 73 (2002) 508–510
Short report
Brine shrimp lethality bioassay of selected Indian medicinal plants R. Padmajaa, P.C. Arunb, D. Prashantha,*, M. Deepakb, A. Amita, M. Anjanab a
Bioassay Unit, Research & Development Centre, Natural Remedies Pvt. Ltd., Plot No. 5B, Veerasandra Indl. Area, Hosur Road, Bangalore 561 229, India b Phytochemistry Laboratory, Research & Development Centre, Natural Remedies Pvt. Ltd., Plot No. 5B, Veerasandra Indl. Area, Hosur Road, Bangalore 561 229, India Received 17 July 2001; accepted in revised form 18 June 2002
Abstract Ethanolic extracts of six Indian medicinal plants, piperine, guggulsterone E and guggulsterone Z were tested for cytotoxicity using brine shrimp lethality test. Piper longum showed most potent cytotoxic activity. Piperine, guggulsterone E and guggulsterone Z showed potent activity with LC50 2.4, 8.9 and 4.9, respectively. 䊚 2002 Elsevier Science B.V. All rights reserved. Keywords: Brine shrimp lethality test; Acorus calamus; Centella asiatica; Centratherum anthelminticum; Mangifera indica; Commiphora mukul; Piper longum; Guggulsterone E; Guggulsterone Z; Piperine
Plant. Acorus calamus L. (Araceae) rhizome, Centella asiatica L. (Apiaceae) whole plant, Centratherum anthelminticum Kuntze. (Asteraceae) seeds, Mangifera indica L. (Anacardiaceae) bark, Commiphora mukul Hook. (Burseraceae) gum resin and Piper longum L. (Piperaceae) fruit. A. calamus, C. asiatica and M. indica were collected from various parts of Bangalore in March 2000. C. anthelminticum was collected from Salem in March 2000. C. mukul was collected from Delhi in *Corresponding author. E-mail address: [email protected] (D. Prashanth). 0367-326X/02/$ - see front matter 䊚 2002 Elsevier Science B.V. All rights reserved. PII: S 0 3 6 7 - 3 2 6 X Ž 0 2 . 0 0 1 8 2 - X
R. Padmaja et al. / Fitoterapia 73 (2002) 508–510
509
September 2000. The samples were authenticated by our Pharmacognosy department where the voucher specimens are preserved. Uses in traditional medicine. A. calamus as an expectorant, carminative, antispasmodic, nervine sedative, C. asiatica to rejuvenate, C. anthelminticum as an anthelmintic, stomachic, diuretic, M. indica as a astringent, in uterine haemorrhage, in haemorhagic diarrhoea, C. mukul in rheumatism, nervous disorders, P. longum in bronchitis, carminative w1x. Previously isolated constituents. A. calamus: sesquiterpenes, C. asiatica: saponins, C. anthelminticum: triterpenes and sterols, M. indica: xanthones, sterols, tannins, P. longum: alkaloids, C. mukul: steroids and diterpenoids w2,3x. Tested material. Soxhlet ethanolic extracts of A. calamus, C. asiatica, C. anthelminticum, M. indica, C. mukul and P. longum (yields on dried weight: 17.89%, 12.0%, 23.94%, 10.4%, 30%, 21.9%, respectively). Preliminary phytochemical screening w4–6x gave positive tests for tannins, b-asarone and alkaloids (A. calamus), saponins, sterols, tannins and asiaticoside (C. asiatica), fats, tannins and sterols (C. anthelminticum), tannins and mangiferin (M. indica), steroids (C. mukul), alkaloids (P. longum). Guggulsterone E w7x and Z w8x were isolated from C. mukul (yields: 0.12% and 0.025%, respectively). Piperine (98%) was purchased from Mys. Plant Lipids Ltd. Cochin, India. Studied activity. Cytotoxicity using Brine shrimp lethality test w9x. Artemia salina cysts (obtained from Laboratory of Aquaculture and Artemia Reference Centre, Table 1 Cytotoxic activity of Indian plant extractsa Tested material
Concentration tested (mgyml)
LC50 (24 h)
Centratherum anthelminticum (seed) Acorus calamus (rhizome) Centella asiatica (whole plant) Mangifera indica (bark) Piper longum (fruit) Commiphora mukul (gum resin) Guggulsterone E Guggulsterone Z Piperine Potassium dichromateb
200, 400, 800 100, 200, 400 100, 500, 1000 200, 400, 800 1, 5, 10 50, 100, 500 1, 5, 10 1, 5, 10 1, 2, 3 10, 30, 50
473.4 (365.6–641.3) 217.0 (143.8–350.8) )1000 576.9 (470.0–716.9) 6.9 (4.2–16.8) 325.7 (198.00–792.9) 8.9 (5.87–39.065) 4.9 (2.95–7.55) 2.4 (1.9–3.3) 28.7 (19.9–36.5)
a b
All determinations were done in triplicate, 95% confidence limits in parentheses. Reference standard.
510
R. Padmaja et al. / Fitoterapia 73 (2002) 508–510
University of Ghent, Belgium) were used. Potassium dichromate was used as reference standard w9x. LC50 (24 h) were determined using Finney program received from Prof. McLaughlin, Purdue University, USA.
Results. Reported in Table 1. Conclusions. Six Indian medicinal plants (selected randomly, mainly based on their availability and apart from their traditional uses) were tested for cytotoxic activity using brine shrimp lethality test. All the extracts showed significant cytotoxic activity except C. asiatica. Guggulsterones E, Z and piperine showed potent cytotoxic activity. Acknowledgments The authors are thankful to Mr R.K. Agarwal, Chairman, Mys. Natural Remedies Pvt. Ltd., Bangalore for his encouragement and Mrs T.S. Geetha for secretarial assistance. References w1x A.K. Nadkarni, Indian materia medica, vol. 1,, Bombay, Popular Prakashan, 1993. w2x A. Husain, O.P. Virmani, S.P. Popli, L.N. Misra, M.M. Gupta, G.N. Srivastava, Z. Abraham, A.K. Singh, Dictionary of Indian medicinal plants, CIMAP, Lucknow, India, 1992. w3x Sathyavathi GV, Ashok KG, Neeraj T. Medicinal plants of India, Indian Council for Medicinal Research 1987: Vol. 2. w4x E.G. Trease, W.C. Evans, Text book of pharmacognosy, 13th ed, Bailliere Tindall, London, 1989, p. 546. w5x C.K. Kokate, A.P. Purohit, S.B. Gokhale, Pharmacognosy, 4th ed, Pune, Nirali Prakashan, 1996, p. 123. w6x H. Wagner, S. Bladt, Plant drug analysis, 2nd ed, Springer-Verlag, New York, 1996. w7x R.B. Arora, V. Kappor, S.K. Gupta, R.C. Sharma, Ind J Exp Biol 9 (3) (1971) 403. w8x S. Nityananda, N.K. Kapoor, Ind J Exp Biol 11 (1973) 395. w9x T.W. Sam, in: S.M. Colegate, R.J. Molyneux (Eds.), Bioactive natural products, CRC Press Inc, USA, 1993, p. 441.
Short report
Brine shrimp lethality bioassay of selected Indian medicinal plants R. Padmajaa, P.C. Arunb, D. Prashantha,*, M. Deepakb, A. Amita, M. Anjanab a
Bioassay Unit, Research & Development Centre, Natural Remedies Pvt. Ltd., Plot No. 5B, Veerasandra Indl. Area, Hosur Road, Bangalore 561 229, India b Phytochemistry Laboratory, Research & Development Centre, Natural Remedies Pvt. Ltd., Plot No. 5B, Veerasandra Indl. Area, Hosur Road, Bangalore 561 229, India Received 17 July 2001; accepted in revised form 18 June 2002
Abstract Ethanolic extracts of six Indian medicinal plants, piperine, guggulsterone E and guggulsterone Z were tested for cytotoxicity using brine shrimp lethality test. Piper longum showed most potent cytotoxic activity. Piperine, guggulsterone E and guggulsterone Z showed potent activity with LC50 2.4, 8.9 and 4.9, respectively. 䊚 2002 Elsevier Science B.V. All rights reserved. Keywords: Brine shrimp lethality test; Acorus calamus; Centella asiatica; Centratherum anthelminticum; Mangifera indica; Commiphora mukul; Piper longum; Guggulsterone E; Guggulsterone Z; Piperine
Plant. Acorus calamus L. (Araceae) rhizome, Centella asiatica L. (Apiaceae) whole plant, Centratherum anthelminticum Kuntze. (Asteraceae) seeds, Mangifera indica L. (Anacardiaceae) bark, Commiphora mukul Hook. (Burseraceae) gum resin and Piper longum L. (Piperaceae) fruit. A. calamus, C. asiatica and M. indica were collected from various parts of Bangalore in March 2000. C. anthelminticum was collected from Salem in March 2000. C. mukul was collected from Delhi in *Corresponding author. E-mail address: [email protected] (D. Prashanth). 0367-326X/02/$ - see front matter 䊚 2002 Elsevier Science B.V. All rights reserved. PII: S 0 3 6 7 - 3 2 6 X Ž 0 2 . 0 0 1 8 2 - X
R. Padmaja et al. / Fitoterapia 73 (2002) 508–510
509
September 2000. The samples were authenticated by our Pharmacognosy department where the voucher specimens are preserved. Uses in traditional medicine. A. calamus as an expectorant, carminative, antispasmodic, nervine sedative, C. asiatica to rejuvenate, C. anthelminticum as an anthelmintic, stomachic, diuretic, M. indica as a astringent, in uterine haemorrhage, in haemorhagic diarrhoea, C. mukul in rheumatism, nervous disorders, P. longum in bronchitis, carminative w1x. Previously isolated constituents. A. calamus: sesquiterpenes, C. asiatica: saponins, C. anthelminticum: triterpenes and sterols, M. indica: xanthones, sterols, tannins, P. longum: alkaloids, C. mukul: steroids and diterpenoids w2,3x. Tested material. Soxhlet ethanolic extracts of A. calamus, C. asiatica, C. anthelminticum, M. indica, C. mukul and P. longum (yields on dried weight: 17.89%, 12.0%, 23.94%, 10.4%, 30%, 21.9%, respectively). Preliminary phytochemical screening w4–6x gave positive tests for tannins, b-asarone and alkaloids (A. calamus), saponins, sterols, tannins and asiaticoside (C. asiatica), fats, tannins and sterols (C. anthelminticum), tannins and mangiferin (M. indica), steroids (C. mukul), alkaloids (P. longum). Guggulsterone E w7x and Z w8x were isolated from C. mukul (yields: 0.12% and 0.025%, respectively). Piperine (98%) was purchased from Mys. Plant Lipids Ltd. Cochin, India. Studied activity. Cytotoxicity using Brine shrimp lethality test w9x. Artemia salina cysts (obtained from Laboratory of Aquaculture and Artemia Reference Centre, Table 1 Cytotoxic activity of Indian plant extractsa Tested material
Concentration tested (mgyml)
LC50 (24 h)
Centratherum anthelminticum (seed) Acorus calamus (rhizome) Centella asiatica (whole plant) Mangifera indica (bark) Piper longum (fruit) Commiphora mukul (gum resin) Guggulsterone E Guggulsterone Z Piperine Potassium dichromateb
200, 400, 800 100, 200, 400 100, 500, 1000 200, 400, 800 1, 5, 10 50, 100, 500 1, 5, 10 1, 5, 10 1, 2, 3 10, 30, 50
473.4 (365.6–641.3) 217.0 (143.8–350.8) )1000 576.9 (470.0–716.9) 6.9 (4.2–16.8) 325.7 (198.00–792.9) 8.9 (5.87–39.065) 4.9 (2.95–7.55) 2.4 (1.9–3.3) 28.7 (19.9–36.5)
a b
All determinations were done in triplicate, 95% confidence limits in parentheses. Reference standard.
510
R. Padmaja et al. / Fitoterapia 73 (2002) 508–510
University of Ghent, Belgium) were used. Potassium dichromate was used as reference standard w9x. LC50 (24 h) were determined using Finney program received from Prof. McLaughlin, Purdue University, USA.
Results. Reported in Table 1. Conclusions. Six Indian medicinal plants (selected randomly, mainly based on their availability and apart from their traditional uses) were tested for cytotoxic activity using brine shrimp lethality test. All the extracts showed significant cytotoxic activity except C. asiatica. Guggulsterones E, Z and piperine showed potent cytotoxic activity. Acknowledgments The authors are thankful to Mr R.K. Agarwal, Chairman, Mys. Natural Remedies Pvt. Ltd., Bangalore for his encouragement and Mrs T.S. Geetha for secretarial assistance. References w1x A.K. Nadkarni, Indian materia medica, vol. 1,, Bombay, Popular Prakashan, 1993. w2x A. Husain, O.P. Virmani, S.P. Popli, L.N. Misra, M.M. Gupta, G.N. Srivastava, Z. Abraham, A.K. Singh, Dictionary of Indian medicinal plants, CIMAP, Lucknow, India, 1992. w3x Sathyavathi GV, Ashok KG, Neeraj T. Medicinal plants of India, Indian Council for Medicinal Research 1987: Vol. 2. w4x E.G. Trease, W.C. Evans, Text book of pharmacognosy, 13th ed, Bailliere Tindall, London, 1989, p. 546. w5x C.K. Kokate, A.P. Purohit, S.B. Gokhale, Pharmacognosy, 4th ed, Pune, Nirali Prakashan, 1996, p. 123. w6x H. Wagner, S. Bladt, Plant drug analysis, 2nd ed, Springer-Verlag, New York, 1996. w7x R.B. Arora, V. Kappor, S.K. Gupta, R.C. Sharma, Ind J Exp Biol 9 (3) (1971) 403. w8x S. Nityananda, N.K. Kapoor, Ind J Exp Biol 11 (1973) 395. w9x T.W. Sam, in: S.M. Colegate, R.J. Molyneux (Eds.), Bioactive natural products, CRC Press Inc, USA, 1993, p. 441.