Screening of Different Artemisia spp. from Western Ghats of Maharashtra for an Anti-Malarial Compound—Artemisinin


Artemisinin, an endoperoxide sesquiterpene lactone has proven effective in treating drug resistant cases of malaria and cancer. Artemisia annua [sweet wormwood] is the sole source for artemisinin production in many countries. To counter the low content in leaves and costly chemical synthesis process in India, alternative ways to produce artemisinin have been sought. In current study, we collected A. pallens, A. japonica and A. nilagirica from Western Ghats of Maharashtra, India and analyzed artemisinin content. Samples were extracted from leaves and florets in various extraction conditions and analyzed using different chromatographic techniques. Thin layer chromatography (TLC) and high performance thin layer chromatography (HPTLC) analysis showed the presence of compound with endoperoxide linkage in A. nilagirica. High performance liquid chromatography (HPLC) analysis showed the detection of artemisinin in methylene dichloride florets extract of A. japonica, but the concentration was too low [1.3 mg/g dry wt.] for further analyses. Gas chromatography/mass spectrometry (GC/MS) analysis identified structurally important components in the A. nilagirica ethyl acetate extract which explores the biosynthetic pathway of artemisinin from its most important precursor amorpha-4,11-diene. This is the first report of chromatographic screening of these Indian varieties of Artemisia spp. for artemisinin content.

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Shukla, V. , Pala, Z. , Alok, A. and Desai, N. (2015) Screening of Different Artemisia spp. from Western Ghats of Maharashtra for an Anti-Malarial Compound—Artemisinin. American Journal of Plant Sciences, 6, 1619-1632. doi: 10.4236/ajps.2015.69162.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Mcvaugh, R. (1984) Flora Novo-Galiciana: A Descriptive Account of the Vascular Plants of Western Mexico Vol. 12. Anderson, W.R., Ed., University of Michigun Press, Ann Arbor.
[2] Singh, A., Kaul, V.K., Mahajan, V.P., Singh, A., Mishra, L.N., Thakur, R.S. and Hussain, A. (1986) Introduction of Artemisia annua in India and Isolation of Artemisinin, a Promising Anti Malarial Drug. Indian Journal of Pharmaceutical Sciences, 48, 137-138.
[3] Rathore, D., McCutchan, T.F., Sullivan, M. and Kumar, S. (2005) Antimalarial Drugs: Current Status and New Developments. Expert Opinion on Investigational Drugs, 14, 871-883.
[4] Acton, N., Klayman, D.L. and Rollman, I.J. (1985) Reductive Electrochemical HPLC Assay for Artemisinin [Quinghaosu]. Planta Medica, 51, 445-446.
[5] Charles, D.J., Simon, J.E., Wood, K.V. and Heinstein, P. (1990) Germplasm Variation in Artemisinin Content of Artemisia annua Using Alternative Method of Artemisinin Analysis from Crude Plant Extract. Journal of Natural Products, 53, 157-160.
[6] Duke, S.O. and Paul, R.N. (1993) Development and Fine Structure of Glandular Trichomes of Artemisia annua L. International Journal of Plant Sciences, 154, 107-118.
[7] Martinez, B.C. and Staba, J. (1988) The Production of Artemisinin in Artemisia annua L. Tissue Cultures. Advances in Cell Culture, 6, 69-87.
[8] Ramachandra, R.S. and Ravishankar, G.A. (2002) Plant Cell Cultures: Chemical Factories of Secondary Metabolite. Biotechnology Advances, 20, 101-153.
[9] Li, M.Y., Jiang, F.S., Yu, X.L. and Miao, Z.Q. (2015) Engineering Isoprenoid Biosynthesis in Artemisia annua L. for the Production of Taxadiene: A Key Intermediate of Taxol. BioMed Research International, 2015, Article ID: 504932.
[10] Putalun, W., Luealon, W., De-Eknamkul, W., Tanaka, H. and Shoyama, Y. (2007) Improvement of Artemisinin Production by Chitosan in Hairy Root Cultures of Artemisia annua L. Biotechnology Letters, 29, 1143-1146.
[11] Paniego, N.B. and Giulietti, A.M. (1996) Artemisinin Production by Artemisia annua L.-Transformed Organ Cultures. Enzymes and Microbial Technologies, 18, 526-530.
[12] White, N.J. (2008) Qinghaosu (Artemisinin): The Price of Success. Science, 320, 330-334.
[13] Laughlin, J.C. (1994) Agricultural Production of Artemisinin—A Review. Transactions of the Royal Society of Tropical Medicine and Hygiene, 88, 21-22.
[14] Deblays, N., Benakis, A. and Collet, G. (1993) Selection and Breeding of High Artemisinin (Qinghaosu) Yielding Strains of Artemisia annua. Acta Horticulturae, 330, 203-206.
[15] Woerdenbag, H.J., Pras, N., Chan, N.G., Bang, B.T., Bos, R., van Uden, W., Van, Y.P., Van Boi, N., Batterman, S. and Luqt, C.B. (1994) Artemisinin, Related Sesquiterpenes, and Essential Oil in Artemisia annua during a Vegetation Period in Vietnam. Planta Medica, 60, 272-275.
[16] Haynes, R.K. (2006) From Artemisinin to New Artemisinin Antimalarials: Biosynthesis, Extraction, Old and New Derivatives, Stereochemistry and Medicinal Chemistry Requirements. Current Topics in Medicinal Chemistry, 6, 509-537.
[17] Lin, A.J., Klayman, D.L., Hoch, J.M., Silverton, J.V. and George, C.F. (1985) Thermal Rearrangement and Decomposition Products of Artemisinin (Qinghauso). The Journal of Organic Chemistry, 50, 4504-4508.
[18] Theoharides, A.D., Smyth, M.H., Ashmore, A.W., Halverson, J.M., Zhou, Z.M., Rider, W.E. and Lin, A.J. (1988) Determination of Dihydroqinghaosu in Blood by Pyrolysis Gas Chromatography/Mass Spectrometry. Analytical Chemistry, 60, 115-120.
[19] Ferriera, J.F. and Gonzalez, J.M. (2009) Analysis of Underivatized Artemisinin and Related Sesquiterpene Lactones by High-Performance Liquid Chromatography with Ultraviolet Detection. Phytochemical Analysis, 20, 91-97.
[20] Bouwmeester, H.J., Wallart, T.E., Jannsen, M.H., Van Loo, B., Jansen, B.J., Posthumus, M.A., Schmidt, C.O., De Kraker, J.W., Konig, W.A. and Franssen, M.C. (1999) Amorpha-4,11-diene Synthase Catalyses the First Probable Step in Artemisinin Biosynthesis. Phytochemistry, 52, 843-854.
[21] Brown, G.D. (2010) The Biosynthesis of Artemisinin (Qinghauso) and the Phytochemistry of Artemisia annua L. (Quinghao). Molecules, 15, 7603-7698.
[22] Starks, C.M., Black, K., Chappell, J. and Noel, J.P. (1997) Structural Basis for Cyclic Terpene Biosynthesis by Tobacco 5-Epi-Aristocholene Synthase. Science, 277, 1815-1820.
[23] Colby, S.M., Alonso, W.A., Katahira, E.J., Mcgarvey, D.J. and Croteau, R. (1993) 4S-Limonene Synthase from the Oil Glands of Spearmint (Mentha spicata): cDNA Isolation, Characterization and the Bacterial Expression of the Catalytically Active Monoterpene Cyclise. The Journal of Biological Chemistry, 268, 23016-23024.
[24] Desjardins, A.E., Hohn, T.M. and Mccormick, S.P. (1993) Trichothecene Biosynthesis in Fusarium Species: Chemistry, Genetics and Significance. Microbiology Reviews, 57, 595-604.
[25] Hohn, T.M. and Beremand, P.D. (1989) Isolation and Nucleotide Sequence of a Sequiterpene Cyclase Gene from the Trichothecene-Producing Fungus Fusarium sporotrichioides. Gene, 79, 131-138.
[26] Proctor, R.H. and Hohn, T.M. (1993) Aristolochene Synthase: Isolation, Characterization and Bacterial Expression of a Sesquiterpenoid Biosynthetic Gene (Aril) from Penicillium roqueforti. The Journal of Biological Chemistry, 268, 4543-4548.
[27] Setzer, W.N. (2008) Germacrene D Cyclization: An Ab Initio Investigation. International Journal of Molecular Sciences, 9, 89-97.
[28] Yoshihara, K., Ohta, Y., Sakai, T. and Hirose, Y. (1969) Germacrene D: A Key Intermediate of Cadinene Compounds and Bourbonenes. Tetrahedron Letters, 27, 2263-2264.
[29] Williams, D.C., McGarvey, D.J., Katahira, E.J. and Croteau, R. (1998) Truncation of Limonene Synthase Preprotein Provides a Fully Active ‘Pseudomature’ Form of This Monoterpene Cyclase and Reveals the Function of the Amino-Terminal Arginine Pair. Biochemistry, 37, 12213-12220.
[30] Cane, D.E., Yang, G., Xue, Q. and Shim, J.H. (1995) Trichodiene Synthase. Substrate Specificity and Inhibition. Biochemistry, 34, 2471-2479.

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