Volume 3 • Issue 2 • July-September 2010
International Journal of Pharmaceutical Sciences and Nanotechnology
Volume 3 • Issue 2 • July - September 2010
Production of Nitidine from Callus Cultures of Toddalia asiatica M. Rajkumar, R.H. Chandra, C. Veeresham
Kakatiya University, Warangal, India.
Toddalia asiatica(Rutaceae) is a woody liana used traditionally in the treatment of malaria, sprains, cough, fever, neuralgia, epilepsy, dyspepsia and other disease conditions. The extracts of the plant have been reported to have anticancer and anti-HIV activities. These activities are mainly attributed to benzophenanthridine alkaloid i.e., nitidine. The yield of the nitidine in intact plant is too low (0.002% w/w). Plant tissue culture has been used as an alternative method for increasing the production of secondary metabolites such as nitidine. In similar attempt the present study was carried out to initiate callus cultures from T.asiatica and evaluate further strategies to improve the production of nitidine. The leaves of T. asiatica were incubated in MS medium supplemented with different combinations of growth hormones. MS medium containing NAA (2mg/L) and Kinetin (1mg/L) was found to yield good friable callus after six weeks of incubation. The pooled callus was extracted and observed for the presence of nitidine. The identification was done by TLC and HPLC by using the standard sample. The callus was found to produce 0.026% w/w of nitidine which was 10 fold more than that of intact plant. The conditions required for growth of callus and production of nitidine were optimized and it is the first report.
KEY WORDS: Toddalia asiatica; Nitidine; Plant tissue culture; production kinetics Introduction Plants represent an unlimited source of natural products. Many of the recently detected phytochemicals exhibit remarkable bioactivities, ranging from anticancer activity, phosphodiesterase inhibition to cytotoxicity against HIVinfected cells. Toddalia asiatica (Linn.) Lamk (Rutaceae) has been in folklore use in India and China from 18th century. T. asiatica is commonly known as Lopez root, forest pepper and wild orange tree (Molmoori et al., 2008). It is an evergreen climber (woody Liana) with rambling stems. It is native of tropical Asia from India and SriLanka to Malaysia; also available in Mauritius and Madagaskar. It is also found in Sumatra, Java, China, Philippines, tropical Africa and Mascarene Islands. It is found almost throughout India up to an altitude of 2500m. It is common in the Nilgiri and Palni Hills and in the shrub forests of Orissa and Andhra Pradesh (Rastogi et al., 1998). The root bark of T. asiatica is a rich source of active constituents belonging to various classes of secondary metabolites * For correspondence: C. Veeresham, Mobile: 09849129584 Email: email@example.com
which include benzophenanthridine, quinoline, protoberberine alkaloids, coumarins, biscoumarins, furanocoumarins, benzopyrans, terpenoids, cyclohexylamides. The important alkaloids include nitidine, magnoflorine, chelerythrine, fagaronine, avicine, decarine, norchelerythrine, dihydronitidine, dihydroavicine, 8-methoxydihydro chelerythrine, skimmianine, berberine, etc. The coumarins include toddanone, toddalenol, toddalenone, toddasin, toddasiatin, toddalosin, toddacoumaquinone, toddacoumalone, aculeatin, pimpinelline, isopimpinelline, toddalolactone (Sharma et al., 1979; Combes et al., 1984; Ishii et al., 1991; Tsai et al., 1997). The plant has characteristic pungent taste. It is used in sprains, contusions, intercostal neuralgia, cough, malaria, dysentery, gastralgia, poisonous snake bites and furuncle infections. Fresh bark of the root is used in the treatment of hill fever. The root is used as dental analgesic. It is also used in odontalgia, paralysis, intermittent fevers, dyspepsia, colic, flatulence, bronchitis,...