Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong>Vol. 5 (2) 1084-1097 April 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1097Implantable Polymers. J. Pharm. Sci.85(12): 1341-1345.38. Joseph, K. (1990). Pulsed <strong>and</strong> SelfregulatedDrug Delivery. CRC press.39. Mendoza, V. L. <strong>and</strong> Vachet, R. W. (2009).Probing Protein Structure by Amino Acid-Specific Covalent Labeling <strong>and</strong> MassSpectrometry. Mass Spectrom Rev. ; 28(5):785–815.40. Tuesca, A. D, Reiff, C., Joseph, J.I. <strong>and</strong>Lowman, A. M. (2009). Synthesis,Characterization <strong>and</strong> In Vivo Efficacy <strong>of</strong>PEGylated Insulin for Oral Delivery withComplexation Hydrogels. PharmaceuticalResearch, 26 (3): 727-739.41. Crommelin, D. J. A., Storm, G., Verrijk, R.,Leede, L. D., Jiskoot, W. <strong>and</strong> Hennink, W.E. (2003). Shifting paradigms:biopharmaceuticals versus low molecularweight drugs. Int. J. Pharm. 266(1-2): 3-16.42. Vasir, J. K., Reddy, M. K. <strong>and</strong> Labhasetwar,V. D. (2005). Nanosystems in DrugTargeting: Opportunities <strong>and</strong> Challenges.Current Nanoscience, 1: 47-64.43. Chappell, J., Leitner, J. W., Solomon, S.,Golovchenko, I., Goalstone, M. L. <strong>and</strong>Draznin B. (2001). Effect <strong>of</strong> insulin on cellcycle progression in MCF-7 breast cancercells. J. Biol. Chem. 276(41): 38023-38028.44. Kubik, T., Bogunia-Kubik, K. <strong>and</strong> Sugisaka,M. (2005). Nanotechnology on Duty inMedical Applications. CurrentPharmaceutical <strong>Biotechnology</strong>, 6: 17-33.45. Liu, J., Zhang, S. M., Chen, P. P., Cheng,L., Zhou, W., Tang, W. X., Chen, Z. W.<strong>and</strong> Ke, C. M. (2007). Controlled release<strong>of</strong> insulin from PLGA nanoparticlesembedded within PVA hydrogels. J MaterSci: Mater Med 18:2205–2210.46. Kawashima, Y., Yamamoto, H., Takeuchi,H., Fujioka, S. <strong>and</strong> Hino T. (1999).Pulmonary delivery <strong>of</strong> insulin with nebulizedDL- lactide / glycolide copolymer (PLGA)nanosphere to prolong hypoglycemic effect,J. Control. Rel. 62: 279-287.Mahmoud et al
Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong>Vol. 5 (2) 1098-1103 April 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1098A Simple <strong>and</strong> Rapid Method for Isolation <strong>of</strong> Alternaric Acid fromAlternaria solaniS.J. Patel*, R.B. Subramanian <strong>and</strong> Y.S. JhaB. R. Doshi School <strong>of</strong> Biosciences, Lab. No.103 B, Bakrol Vadtal Road, Satellite Campus,Sardar Patel University, Vallabh Vidhyanagar 388120, India*For correspondence - suchitapatel5@yahoo.comAbstractAlternaric acid is the major toxin <strong>of</strong>Alternaria solani, the causative agent <strong>of</strong> earlyblight <strong>of</strong> tomato. A simple <strong>and</strong> rapid method hasbeen developed for the isolation <strong>of</strong> alternaric acidin crystalline form from the fungal culture filtrate(FCF) with ethanol <strong>and</strong> carbon tetrachloride. Thecrystalline form <strong>of</strong> alternaric acid obtained wassubjected to High Performance Thin LayerChromatography (HPTLC), spectrophotometricanalysis <strong>and</strong> detached leaf assay to confirm thepurity <strong>and</strong> biological activity <strong>of</strong> alternaric acid.This one step protocol is superior to earlierreported methods <strong>and</strong> also gives higher yield.Keywords: Acid, Blight, Tomato, Toxin, TLCIntroductionTomato (Lycopersicon esculentumMill.) belongs to the family Solanaceae.Worldwide, it is the second most consumedvegetable after potato (1). Early blight <strong>of</strong> tomatocaused by the necrotrophic fungus Alternariasolani Sorauer is perhaps the most common foliardisease <strong>of</strong> tomatoes. In USA, Australia, Israel,UK, <strong>and</strong> India, significant reductions in yield (35up to 78%) due to early blight (EB) have beenobserved (2, 3, 4). The symptoms occur on theleaves where circular <strong>and</strong> concentric lesions areproduced. Early blight reduces the photosyntheticarea <strong>and</strong>, in severe cases, can defoliate plants.The fungus survives between crops as myceliaor conidia in soil, plant debris <strong>and</strong> seed (5). Thespores are transported by water, wind, insects,other animals including man, <strong>and</strong> machinery.Eleven toxins have been identified in culturefiltrates <strong>of</strong> A. solani (6). Among these, alternaricacid <strong>and</strong> solanapyrone A, B, <strong>and</strong> C are able toinduce necrotic symptoms similar to EB symptoms(6). Out <strong>of</strong> these, alternaric acid is one <strong>of</strong> themajor toxins found in the filtrates (7) which causeschlorosis <strong>and</strong> necrosis <strong>and</strong> therefore plays a majorrole in early blight symptoms <strong>and</strong> defoliation (8).Alternaric acid does not cause phytotoxicity whensprayed alone on tomato leaves, but it enhancesthe infection process <strong>and</strong> the development <strong>of</strong>necrotic symptoms when added to A. solani sporesuspensions. Alternaric acid was previouslycrystallized by Brian et al. (9). Stoessl <strong>and</strong>Stothers (10) developed a method for extraction<strong>of</strong> alternaric acid from fungal culture filtrate.Previously, Matern et al. (11) tried toisolate alternaric acid from crude FCF. Theprocedure is time consuming <strong>and</strong> the results werenot reliable. Maiero et al. (12) made an attemptto isolate alternaric acid in bulk quantities forscreening <strong>of</strong> resistant varieties, but wereunsuccessful. St<strong>and</strong>ard alternaric acid or alternaricacid in pure form are difficult to obtainIsolation <strong>of</strong> Alternaric acid