Total views : 107

Rationale for Water Extraction of Lipophilic Azadirachtin ( Tetranor Triterpenoid) from Neem Seeds - A Conundrum

Affiliations

  • Department of Chemical Sciences, Vittal Mallya Scientific Research Foundation, Bangalore-560062, India
  • Department of Biological Sciences, Vittal Mallya Scientific Research Foundation, Bangalore-560062

Abstract


Objectives: To extract lipophilic Azadirachtin from seed kernels of Azadirecta indica (Neem) with water and the rationale behind it as its solubility in water is very negligible (0.26 gm/lit of water).Methods: Neem seeds obtained from local market were dehulled to obtain kernels which were cold pressed to remove oil and extracted with four volumes of water at room temperature. Azadirachtin content in cold pressed neem kernel was determined using High Performance Liquid Chromatography (HPLC) with reverse phase silica column (RP18).Proteincontentinaqueousextractwasdeterminedby micro-Kjeldahl’s method and sugars by acid hydrolysis, permethylation and Gas Chromatography (GC). Findings: Aqueous extract of neem seed kernel was found to contain about 670 ppm of azadirachtin (AZA). A small portion of this aqueous extract when lyophilized yielded a hygroscopic material showing the presence of 4% AZA (HPLC method), 25% proteins (micro-Kjeldahl’s method) and about 50% of carbohydrates and other aliphatic compounds. The aqueous solution when re-extracted into ethyl acetate followed by precipitation using n-hexane yielded azadirachtin of 40% purity.Application: Lipophilic azadirachtins get extracted into water may be due to micelle formation of polysaccharides and proteins that cooccur in neem seed kernel.This hypothesis has been substantiated by addition of external azadirachtin to aqueous extract of neem seed kernel thereby enhancing its solubility from initial 670 ppm by more than two fold to a level of 1800 ppm. To obtain azadirachtin of about 40% purity directly from neem seed kernels would be possible only when first extracted with water.

Keywords

Azadirachtin, lipophilic, neem seed kernel, polysaccharides, proteins

Full Text:

 |  (PDF views: 121)

References


  • Schmutterer H. The Neem Tree. Germany:VCH Weinheim, Publ; 1995. p. 1–23. Crossref
  • Heinz R. Insecticides of Plant Origin. USA: ACS Symposium Series, ACS Publ; Chapter 11. 1989. 150–63.
  • Rao AR, Kumar SSV, Paramsivam TR. Kamalakshi S, Parasuraman AR, Shanta B. Study of antiviral activity of leaves of margosa tree on vaccinia and vriola viruses- a preliminary report. Indian Journal of Medicinal Research.
  • ; 57: 495–502. PMid:4309947
  • Gogate SS, Marathe A. Antiviral effects of neem leaf (Azadirachta indica Juss.) Journal of Research and Education in Indian Medicine. 1989; 8(1):1–7.
  • padhyay SN, Dhavan S, Garg SN, Wali N, Tusker L, Anderson DJ. Immunomodulatory properties of neem. World Neem Conference, Bangalore, India. 1993; 4(3): 1–10.
  • Amer H, Helmy H, Taie HAA. In vitro antitumor and antiviral activities of herbal plant seeds and leaves extracts.
  • International Journal of Academy Research. 2010; 2: 47–51.
  • Miller JA, Chamberlain WF. Azadirachtin as a larvicide against the horn fly, stable fly, and house fly (Diptera: Muscidae). Journal of Economic Entomology. 1989; 82: 1375–78. Crossref PMid:2600264
  • Schroeder R, Nakanishi KA. Simplified isolation procedure for azadiracthin. Journal of Natural Products. 1987; 50: 241–4. Crossref PMid:3655797
  • Henderson R, McCrindle R, Overtone KH, Metera A. Salannin. Tetrahedron Letters. 1964; 52: 3969–74. Crossref
  • Zanno PR, Maira E, Nakanishi K, Elder DL. Structure of the insect phagorepellent azadirachtin. Application of PRFT/ CWD [partially relaxed Fourier transform/continuous wave decoupling] carbon-13 nuclear magnetic resonance. Journal of American Chemical Society.1975; 97(7): 1975– 77. Crossref PMid:1133406
  • Ermel K, Pahlich I, Schmutterer H. Proceedings of the Second International.Neem Conefence (Eds Schumutter,H. and Aschier, K.R.S) GTZ. Germany: 1984.p. 1–83.
  • Ermel K, Pahlich I, Schmutterer, H. Proceedings of International. Neem Conference. Nairobi, Kainya: 1986.p.
  • –84.
  • Rengasamy S, Kaushik N, Kumar J, Kaul O, Parmar BS. In world Neem Conference (Ed. Singh,R.P), New Delhi: Oxford and IBH; 1993.p. 207–17.
  • Sidhu OP, Kumar V, Behl HM. Variability in neem (Azadirachta indica) with respect to azadirachtin content.
  • Journal of Agricultural Food Chemistry. 2003; 51: 910–15. Crossref PMid:12568548
  • Bally ISE, Ruddle L, Simpson B. Azadirachtin levels in neem seeds grown in Australia. Abstract International Neem Conference, Queensland, Australia: 1996.p. 1–17.
  • Nutan K, Gurudevsingh B, Sharma KK, Banerjee SK, Pramilla T. Regional and habitat variability in azadirachtin content of Indian neem (Azadirachta indica A. Jusieu). Current Science. 2007; 92: 1400–06.
  • Nagasampagi. patent no US 5,395,951, 1995. Kleberg. patent no US 5,695,763, 1997.p. 1–43.
  • Butterworth JH, Morgan ED, Percy, GR. The structure of azadirachtin; the functional groups. Journal of the Chemical Society, Perkin Transactions. 1972; 1:. 2445–50. Crossref
  • Uebel EC, Warthen JD, Jacobson M. Preparative ReversedPhase Liquid Chromatographic Isolation of Azadirachtin from Neem Kernels (1).Journal of Liquid Chromatography. 1979; 2(6): 875–882. Crossref
  • Yamasaki RB, Klocke JA, Lee SM, Stone GA, Darlington MV. Isolation and purification of Azadirecta indica seeds using flash chromatography and high performance liquid chromatography. Journal of Chromatography. 1986;, 356: 220–6. Crossref
  • Lindert Z, Craig G, Clovis JS. Preparation of high purity neem seed extracts. Patent no US 5,420,318, 1995.
  • Oshea MJ. Brushett DJ, Pollard DJ. Azadirachtin extraction process. Patent no US 6312738, 2001.
  • Kleeberg H, MGmbH T. Azadirachtin extraction process. Patent no AU 661482, 1991.
  • Shivaprakash S, Kumar AP, Kush A, Reddy GC. Indian Patent Application No. 1907/CHE/2010, 2012.p. 1–827.
  • Daniel Derbie AMS. Thesis. Canada: The University of Manitoba;1999. p. 43.
  • Wang CJ, Dimitry M, Vladimir PJ. Polymeric micelles for delivery of poorly soluble drugs: preparation and anticancer activity in vitro of paclitaxel incorporated into mixed micelles based on Poly(ethylene Glycol)-Lipid conjugate and positively charged lipids. J Drug Target. 2005; 13(1): 73–80.
  • Canu I, Kerek F. A simple and rapid method for the permethylation of carbohydrates. Carbohydrate Research.1984; 131: 209–217. Crossref
  • Francis MF, Cristea M, Francoise MW. Polymeric micelles for oral drug delivery: Why and how. Pure and Applied Chemistry. 2004; 76: 1321–35. Crossref
  • Kumble PS, Annadurai RS, Sreenivas M, Rao AS, Ramdoss CS, Rao PVS. Indian patent 187645, 2002 and US 6635757. 2003.p. 1-2.
  • Liu Y, Chen GS, Lin J. Inclusion complexes of azadirachtin with native and methylated cyclodextrins: solubilization and binding ability. Bioorganic Medicinal Chemistry. 2005; 13(12): 4037–42. Crossref PMid:15911316

Refbacks

  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution 3.0 License.