Bioanalytical Techniques - KopyKitab

Abhilasha ShourieShilpa S. ChapadgaonkarBioanalytical techniques form an integral part of applied biologyand biomedical sciences. The book provides understandingof the concept and working principles of various bioanalyticaltechniques used in biomedical sciences, environmental studies,life sciences, pharmaceutical analysis, molecular biology, andbiotechnological research, as well as the various instrumentsused in these processes. Divided into 12 chapters, the bookprovides a comprehensive account of microscopy, centrifugation,chromatography, electrophoresis, spectroscopy. It also focuses ontwo main topics: radioisotope and immunodiagnostic techniques.Techniques in molecular biology and recombinant DNA technologyhave also been described in detail. Explains analytical instrumentation in a concise manner Provides state-of-the-art sophisticated techniques that wouldbe beneficial to researchers in various fields for experimentation Encourages reader to analytical thinking and practicalapplication of the techniqueBioanalyticalTechniquesAbhilasha ShourieShilpa S. ChapadgaonkarShourie ChapadgaonkarKey FeaturesBioanalytical TechniquesBioanalyticalTechniquesThe Energy and Resources Institute

Abhilasha ShourieShilpa S. ChapadgaonkarThe Energy and Resources Institute

The Energy and Resources Institute, 2015ISBN 978-81-7993-529-3All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, ortransmitted in any form or by any means, electronic, mechanical, photocopying, recording orotherwise, without the prior permission of the publisher.All export rights for this book vest exclusively with The Energy and Resources Institute (TERI).Unauthorized export is a violation of terms of sale and is subject to legal action.Suggested citationShourie, Abhilasha and Shilpa S. Chapadgaonkar. 2015. Bioanalytical Techniques.New Delhi: TERIPublished byThe Energy and Resources Institute (TERI)TERI PressTel. 2468 2100 or 4150 4900Darbari Seth BlockFax 2468 2144 or 2468 2145IHC Complex, Lodhi Road India 91 Delhi (0)11New Delhi – 110 003E-mail teripress@teri.res.inIndiaWebsite www.teriin.orgBookstore https://bookstore.teri.res.inPrinted in India

PrefaceScience and technology by far depend upon experimental procedures involvingone or more analytical methods. Experiments are conducted not only topredict phenomena, but also to validate the results. Analysis of chemical andbiochemical entities is metaphorically the path to achieve the objectives ofresearch in life sciences. In the past few decades, the field of life science haswitnessed rapid advancements through development of highly sophisticated,automated, sensitive, and accurate analytical techniques. A vast range ofanalytical techniques and respective instruments are available, and thereforeit is imperative to understand the principles, limitations, and alternatives of agiven technique in order to apply it effectively to obtain useful results.‘Bioanalytical techniques’ is included as a fundamental paper in most coursesin chemistry, biochemistry, biology, pharmaceutical and clinical sciences,environmental, forensic and materials sciences. In the past 10 years of ourcareer, we have felt a profound need for a comprehensive yet intensive textbookon analytical techniques that would serve the purpose of both students andresearchers. This book has been written to fulfil the need for a text-cumreference in undergraduate and postgraduate level curriculum, providingnecessary information required to demonstrate the concepts of an analyticaltechnique in all its guises.The book emphasizes on imparting profound knowledge that is able tomeet the current throughput screening demands of scientists and researchers.It consists of 12 chapters, encompassing techniques used for biological andbiochemical separation, purification, identification and quantification, all puttogether to construct a compact package. The chapters have been preparedmeticulously using simple yet lucid language. We have included a fairly good

vi Prefacenumber of state-of-the-art sophisticated techniques that would be beneficial toresearchers in various fields for experimentation.Many typical analytical procedures appear in the book as boxed featureswhich give a snapshot of the techniques being widely used contemporarily inresearch. Throughout the book, wherever felt required, extended illustrativeexamples have been incorporated, which are to be read as part of the respectivechapter. The format also encourages the reader to analytical thinking andpractical application of the technique. This book will certainly prove to be aninvaluable reference tool for students, teachers and researchers in the mentionedfields.We express our gratitude to Almighty and all those who have contributed tothe book in any manner. We thank our families for their encouragement andunconditional support. We also thank the editorial team of TERI Press fortheir continuous effort and faith in us.We solicit constructive suggestions from all the readers for further improvementof the content of the book.Dr. Abhilasha ShourieDr. Shilpa S. Chapadgaonkar

ContentsPreface1.GENERAL PRINCIPLES OF ANALYTICAL tal StudiesExperimental ErrorsStatistical Parameters for Validation of an ExperimentSOLUTIONS AND BUFFERS2.12.22.32.42.52.62.73.vIntroductionUnits of ConcentrationThe Concept of pHAcids and BasesHenderson–Hasselbalch EquationDetermination of l BackgroundNature of LightCompound MicroscopeImage Formation in a Light MicroscopePhase Contrast MicroscopyFluorescence MicroscopyElectron Microscopy1127711111216171920203737394143475154

viii Contents4.CELL 5.15.25.35.45.55.66.IntroductionTypes of Chromatographic TechniquesPlanar ChromatographyColumn ChromatographyProtein Purification .IntroductionPrinciples of CentrifugationCentrifuge MachinesCentrifugal SeparationsAnalytical UltracentrifugationCare and Safety of CentrifugesCHROMATOGRAPHIC TECHNIQUES6.16.26.36.46.57.IntroductionBarriers for Cell DisruptionMethods of Cell Disruption—an OverviewMechanical Methods of Cell DisruptionNon-Mechanical Methods of Cell DisruptionCombinations of Cell Disruption MethodsSelection of Cell Disruption MethodsAnalysis of Cell DisruptionIntroductionPrinciples of ElectrophoresisFree Solution ElectrophoresisPaper ElectrophoresisGel ElectrophoresisGel Electrophoresis of ProteinsGel Electrophoresis of Nucleic AcidsCapillary ElectrophoresisSPECTROSCOPY I8.1 Electromagnetic Radiations8.2 Ultraviolet and Visible Light Spectroscopy8.3 Fluorescence 28238

Contents8.4 Atomic Absorption Spectrometry8.5 X-ray Spectroscopy8.6 Circular Dichroism and Optical Rotatory Dispersion9.SPECTROSCOPY II9.19.29.39.49.5IntroductionInfrared SpectroscopyNuclear Magnetic Resonance SpectroscopyElectron Spin Resonance SpectroscopyMass Spectroscopy10. RADIOISOTOPE tion and ApplicationsStructure of Atom and RadioactivityTypes of Radioactive DecayInteraction of Radioactivity with MatterKinetics of Radioactive DecayUnits of RadioactivityDetection and Measurement of RadioactivitySafety Issues and Radio-waste Management11. IMMUNOCHEMICAL TECHNIQUES11.111.211.311.4IntroductionProduction and Purification of AntibodiesImmunoassay TechniquesAdvances in Immunochemical Techniques12. MOLECULAR BIOLOGY Isolation of Nucleic AcidsQualitative and Quantitative Evaluation of Nucleic AcidsNucleic Acid HybridizationRestriction DigestionNucleic Acid SequencingDNA Amplification by Polymerase Chain ReactionFurther ReadingsIndexAbout the 87393397411

Bioanalytical Techniques35%OFFPublisher : TERI PressISBN : 9788179935293Author : Abhilasha ShourieType the URL : http://www.kopykitab.com/product/8493Get this eBook

3. MICROSCOPY 37 3.1 Historical Background 37 3.2 Nature of Light 39 3.3 Compound Microscope 41 3.4 Image Formation in a Light Microscope 43 3.5 Phase Contrast Microscopy 47 3.6 Fluorescence Microscopy 51 3.7 Electron Microscopy 54