CHROMATOGRAPHY ANDITSAPPLICATIONS EditedbySasikumarDhanarasu Chromatography and Its Applications Edited by Sasikumar Dhanarasu Published by InTech Janeza Trdine 9, 51000 Rijeka, Croatia Copyright © 2012 InTech All chapters are Open Access distributed under the Creative Commons Attribution 3.0 license, which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. After this work has been published by InTech, authors have the right to republish it, in whole or part, in any publication of which they are the author, and to make other personal use of the work. Any republication, referencing or personal use of the work must explicitly identify the original source. As for readers, this license allows users to download, copy and build upon published chapters even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. Notice Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book. Publishing Process Manager Vedran Greblo Technical Editor Teodora Smiljanic Cover Designer InTech Design Team First published March, 2012 Printed in Croatia A free online edition of this book is available at www.intechopen.com Additional hard copies can be obtained from orders@intechopen.com Chromatography and Its Applications, Edited by Sasikumar Dhanarasu p. cm. ISBN 978-953-51-0357-8 Contents Preface IX Chapter 1 Column Liquid Chromatography 1 Changming Zhang, Zhanggen Huang and Xiaohang Zhang Chapter 2 Column Chromatography for Terpenoids and Flavonoids 13 Gülçin Saltan Çitoğlu and Özlem Bahadır Acıkara Chapter 3 Chromatographic Separation and Identification of Sildenafil and Yohimbine Analogues Illegally Added in Herbal Supplements 51 Hakan Göker, Maksut Coşkun and Gülgün Ayhan-Kılcıgil Chapter 4 Purification of Marine Bacterial Sialyltransferases and Sialyloligosaccharides 69 Toshiki Mine and Takeshi Yamamoto Chapter 5 Simple Preparation of New Potential Bioactive Nitrogen-Containing Molecules and Their Spectroscopy Analysis 87 Vladimir V. Kouznetsov, Carlos E. Puerto Galvis, Leonor Y. Vargas Méndez and Carlos M. Meléndez Gómez Chapter 6 Wound Healing and Antibacterial Properties of Leaf Essential Oil of Vitex simplicifolia Oliv. from Burkina Faso 109 Magid Abdel Ouoba, Jean Koudou, Noya Some, Sylvin Ouedraogo and Innocent Pierre Guissou Chapter 7 Use of Associated Chromatographic Techniques in Bio-Monitored Isolation of Bioactive Monoterpenoid Indole Alkaloids from Aspidosperma ramiflorum 119 Talita Perez Cantuaria Chierrito, Ananda de Castro Cunha, Luzia Koike, Regina Aparecida Correia Gonçalves and Arildo José Braz de Oliveira VI Contents Chapter 8 Secondary Metabolites 131 Tânia da S. Agostini-Costa, Roberto F. Vieira, Humberto R. Bizzo, Dâmaris Silveira and Marcos A. Gimenes Chapter 9 Biomarkers 165 Yasser M. Moustafa and Rania E. Morsi Chapter 10 Quantification of Antimalarial Quassinoids Neosergeolide and Isobrucein B in Stem and Root Infusions of Picrolemma sprucei Hook F. by HPLC-UV Analysis 187 Rita C. S. Nunomura, Ellen C. C. Silva, Sergio M. Nunomura, Ana C. F. Amaral, Alaíde S. Barreto, Antonio C. Siani and Adrian M. Pohlit Chapter 11 Purification of Peptides from Bacillus Strains with Biological Activity 201 María Antonieta Gordillo and María Cristina Maldonado Preface Chromatography is apowerfulseparationtoolthat isusedinall branchesofscience, and is often the only means of separating components from complex mixtures. The Russian botanist Mikhail Tswett coined the term chromatography in 1906. The first analyticaluseofchromatographywasdescribedbyJamesandMartinin 1952,forthe useofgaschromatographyfortheanalysisoffattyacidmixtures. Awiderangeofchromatographicproceduresmakesuseofdifferencesinsize,binding affinities, charge, and other properties.Many types of chromatography have been developed. These include Column chromatography, High performance liquid chromatography (HPLC), Gas chromatography, Size exclusion chromatography, Ion exchangechromatographyetc. In this book contains more details about the applications of chromatography by various research findings. Each and every topics of this book have included lists of references at the end to provide students and researchers with starting points for independent chromatography explorations. I welcome comments, criticisms, and suggestionsfromstudents,facultyandresearchers. Dr.D.Sasikumar DepartmentofBiochemistry,CollegeofMedicine, UniversityofHail,Hail, KingdomofSaudiArabia [...]... et al (1984) which employs normal phase chromatography columns for the separation procedures and includes multiple (seven) steps respectively as follows; 1 Extraction with alcohol and concentration, 2 Partition between water and dichloromethane, 3 Filtration chromatography, 4 Silica column chromatography, 5 Alumina chromatography 6 Medium pressure silica column chromatography, 7 Preparative HPLC For... compounds can be separated by fractional recrystalization, repeated column chromatography, reversed phase HPLC, chromatography with Sephadex LH-20 or more efficiently by chromatography on NaOAc impregnated silica gel In the following method was described by Jaracz et al (2004) for the isolation of bilobalide and ginkgolides using column chromatography The enriched triterpene trilactone extract was chromatographed... 7.0 with 5 N NaOH After centrifugation supernatant was subjected to XAD-2 column chromatography and eluted with water then MeOH The MeOH eluate was fractioned by multilayer coil counter current chromatography using CHCl3-MeOH-H2O (7:13:18) to yield fifty fractions Fractions 20-30 were rechromatographed on silica gel column chromatography using CHCl3-MeOH (7:1, 5:1, 4:1, 3:1) mixtures Fractions eluted... organic,1987,05-23 [7] Justin D, Fair, Chad M.Kormos, Flash column chromatograms estimated from thin-layer chromatography data Journal of Chromatography A, 2008, 1211:49-54 [8] Davies, Don R., Johnson, Todd M Isolation of three components from spearmint oil: An exercise in column and thin-layer chromatography J Chem Educ 2007,84(2):318-320 [9] B Concho-Grande, M Rodriguez-Comesafia, J.Simal-Gandara,... hexane or gradient elution with mixtures of solvents in increasing polarity leads to successive isolation (Sur, 1991) Additionally, faster techniques of column chromatography such as flash chromatography may be preferred due to conventional column chromatography for separating procedure is time-consuming and frequently gives poor recovery owing to band tailing (Ikan, 1991) The genus Tagetes belongs to... hexane can be preferred due to artifact formation at the raised temperatures (Harborne, 1998) Adsorbtion chromatography on silica gel is the simplest and most effective method for separation of terpenoids and GLC is used commonly for identification as well as isolation of the monoterpenoids Column chromatography is also a valid method for fractionation of monoterpenoids Isocratic elutions with solvents... liquid chromatography (HPLC) to hydrocarbon group-type analysis is characteristic with its high efficiency, high speed, and high sensitivity But HPLC is only suitable for analysis of substances soluble in n-pentane [1] TLC-FID [2-3] method can be also used to analysis the THF-soluble party in asphalt-samples and show great advantages But, the components were combusted during TLC-FID analysis 2 Chromatography. .. 3,7-dimethyl-5-hydroxyoct-1-en-6-one and 3,7-dimethyloct-1,7dien-6-one were obtained by rechromatography of fraction V respectively (Garg & Mehta, 1998) Tagetes patula L another species from this genus allows to the isolation of acyclic monoterpene glycosides Methanolic extract of the flowers was separated on silica gel column chromatography using CHCl3-MeOH mixtures to yield 2-methyl-6-methylen-2,7octadiene... tridentata ssp spiciformis was separated by flash chromatography on silica gel using 19:1 hexane-EtOAc followed by 4:1 hexane-EtOAc except for the oil isolated from A tridentata ssp spiciformis which was flash chromatographed with 9:1 hexane-EtOAc as the second solvent system A tridentata ssp vaseyana essential oil was separated into three major fractions by column chromatography GC analysis of the first chromatographic... with CH2Cl2, dried (MgSO4) and solvent removed under reduced pressure to yield an aromatic green gum which was subjected to gradient (hexaneEtOAc 5 to 100%) column chromatography yielding 32 crude fractions The crude fractions from column chromatography were further purified by repeated prep HPLC, using n-hexaneEtOAc-HOAc in varying proportions, according to the polarity of the crude fraction which . types of chromatography have been developed. These include Column chromatography, High performance liquid chromatography (HPLC), Gas chromatography, Size exclusion chromatography, . Tswett coined the term chromatography in 1906. The first analyticaluseof chromatography wasdescribedbyJamesandMartinin 1952,forthe useofgas chromatography fortheanalysisoffattyacidmixtures. Awiderangeofchromatographicproceduresmakesuseofdifferencesinsize,binding affinities,. CHROMATOGRAPHY ANDITSAPPLICATIONS EditedbySasikumarDhanarasu Chromatography and Its Applications Edited by Sasikumar