logging in or signing up electrophoresis ppt nishit_patel5 Download Post to : URL : Related Presentations : Let's Connect Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Copy embed code: Embed: Flash iPad Dynamic Copy Does not support media & animations Automatically changes to Flash or non-Flash embed WordPress Embed Customize Embed URL: Copy Thumbnail: Copy The presentation is successfully added In Your Favorites. Views: 3704 Category: Science & Tech.. License: All Rights Reserved Like it (1) Dislike it (0) Added: November 10, 2011 This Presentation is Public Favorites: 3 Presentation Description No description available. Comments Posting comment... Premium member Presentation Transcript ELECTROPHORESIS : ELECTROPHORESIS Prepared by : Maulik Oza M.Pharm (Quality Assurance) Roll no : 6 Guided by : Mr.Dharmendrasinh A. Baria Assistant Professor M.Pharm (Pharmaceutical chemistry) Dharmaj Degree Pharmacy College Dharmaj 1ELECTROPHORESIS : ELECTROPHORESIS When a potential difference is applied between the electrodes in a colloidal solution ,colloidal particles are carried to either positive or negative electrode. In short they are electrically charged with reference to dispersion medium. Migration of charged solutes in a liquid medium under an electrical field Many biological molecules have ionisable groups eg. amino acids, proteins, nucleotides, nucleic acids 2Slide 3: Under an electric field charged particles migrate to anode (+) or cathode (-) Migration of individual ions as well as to colloidal aggregate . Rate of travel of the partical depend on Characteristics of the particals. Properties of the electrical field. Temperature. Nature of the suspended medium 3Slide 4: Application for the purification Preparation & measurement. Electrophoresis methods are powerful analytical tool for fractionating component of mixture are aggregate or monodispersed. 4TYPES OF ELECTROPHORESIS : TYPES OF ELECTROPHORESIS FREE SOLUTION ELECTROPHORESIS Moving boundary electrophoresis Density gradient electrophoresis ZONE ELECTROPHORESIS Paper electrophoresis Gel electrophoresis Capillary zone electrophoresis Immuno electrophoresis 5Slide 6: FREE SOLUTION ELECTROPHORESIS Absence of supporting & stabilizing medium ZONE ELECTROPHORESIS Presence of supporting & stabilizing medium eg. Paper 6Principle: : Principle: Movement of charged particles when suspended or dissolved in a polar liquid and the mixture is placed in an electric field. Movement of charged particles in gases, although with modifications the basic concepts could be applied to these suspensions. For most purposes the movement of the ion is translational and carried out under the influence of a constant electric field. 7THEORY : THEORY When a constant electric field is applied across a column of water, current flows and gases evolve at the electrodes hydrogen at the cathode and oxygen at the anode. This phenomenon is known as electrolysis. The experimental basis for these laws of conduction of solutions involved the measurement of conductivity of a solution electrochemical equivalences showing that the conductance of a solution is the product of the number of ion in the solution, the charge carried by each ion, and the velocity or their mobilities u ±. 8Slide 9: Quantitative relationships were developed between the current i carried through the solution by C + gram moles of a univalent anion and the mobilities observed in an electricfield E : i + = u + C + E . (1) The equivalent expression for the cat ion is the same as Eq. (1), and so the total current i is i = i + + i − = ( u + C + + u − C −) E , (2) which can be compared directly with define resistance, E = 1 /k, (3) Where k is the conductance, hence, k = u + C + + u − C − and the conductance is proportional to the velocities of the ions. 9Slide 10: FIGURE Idealized electrophoresis experiment in which boundaries between three salts are formed initially at a and b . After passage of a known quantity of electricity, the boundaries have moved to a and b . 10Slide 11: It by forming a suitable boundary between the ion and the solvent and measuring the velocity of this boundary given in Fig. . A solution of a simple salt, KCl , separates An elementary arrangement for making this experiment is two solutions of other salts, LiCl in the anode compartment and potassium acetate in the cathode compartment. 11Moving boundary electrophoresis : Moving boundary electrophoresis This method allows the charged species to migrate in a free moving solution in absence of a supporting medium. Samples are fractioned in a U shaped tube that has been filled with unstabilised buffer An electrical field is applied by means of electrodes at the ends of the tube Separation takes place as a result of difference in mobilities Bands are obtained which are located by optical systems 12Slide 13: 13Slide 14: Sample can be large ,so long as the boundary,or long edge, between sample & buffer is kept sharply defined. When such a prepation is subjected to electrophoresis, the boundary of each component retains its identity. Component move at rates proportional to their mobilities . As a result, a pattern is developed in which components overlap to make sharply defined steplike contribution to the total solute concentration. Each component within the tube is visualised as a separate cylinder of solute migrating in an independent manner. 14Slide 15: Cylinders move at different rates, so sharp changes in concentration occur at the points where successive fraction overlape . The boundaries of the fraction can be detected by scanning the tube with a refractometer,because refractive index of a solution is directiy proposional to the concentration of the solute. The conc. of each fraction will be calculated as a function of its contribution to the change in refractive index. This is called as moving boundry electrophoresis. The fundamental difference between the two methods is that zone electrophoresis uses an anticonvection medium boundry electrophoresis does not. 15Reference : Reference Encyclopedia of Physical science and Technology Third Edition Analytical Chemistry S. P. Spragg Pages 363-378. Instrumental analysis by b.k . sharma 3 th edition page no c-269-273. 16Slide 17: Thank You 17 You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.