Paper Chromatography Principle slideshare .pptx

Paper Chromatography
Introduction
Principle
Instrument
Methodology
Modes of Paper Chromatography
Application
Advantages
Disadvantages

1. PAPER CHROMATOGRAPHY Presented by SABITRI PRADHAN M.Pharm,GIPS Pharmacology, 1st semester 1

2. CONTENTS INTRODUCTION PRINCIPLE INSTRUMENT METHODOLOGY MODES OF PAPER CHROMATOGRAPHY APPLICATION ADVANTAGE DISADVANTAGES REFERENCE 2

3. INTRODUCTION Paper Chromatography (PC) was first introduced by German scientist Christian Friedrich Schonbein (1865). Paper chromatography is defined as technique in which the analysis of unknown substance is carried out mainly by the flow of solvents on specially designed filter paper. PC is considered to be the simplest and most widely used of the chromatographic techniques because of its applicability to isolation, identification and quantitative determination of organic and inorganic compounds. 3

4. PRINCIPLE The principal of separation is mainly partition rather then absorption cellulose layers in filter paper contains moisture which acts as stationary phase. organic solvents or buffers are used as mobile phases, instead of water as stationary phase other organic solvents can be used by suitable modification. They are two types of paper chromatography: Paper adsorption chromatography. Paper partition chromatography. 4

5. INSTRUMENTATION 5

6. METHODOLOGY 6 A small spot of sample is applied to a strip of chromatography paper about two centimeters away from the base of the plate. This sample is absorbed onto the paper and may form interactions with it. The paper is then dipped into a solvent, such as ethanol or water, taking care that the spot is above the surface of the solvent , and placed in a sealed container. The solvent moves up the paper by capillary action and dissolves the sample mixture, which will then travel up the paper with the solvent solute sample.

7. Contd … 7 Different compounds in the sample mixture travel at different rates . It takes several minutes to several hours. Analysis- Spots corresponding to different compounds may be located by their color, UV light, Ninhydrin or by treatment with iodine vapors . Rf Value = Distance travelled by solute /Distance travelled by solvent

8. MODES OF PAPER CHROMATOGRAPHY Ascending Chromatography (go up)- In this method, the solvent is in pool at the bottom of the vessel in which the paper is supported. It rises up the paper by capillary action against the force of gravity.  Descending Chromatography (a downward slope)- In this method , the solvent is kept in a trough at the top of the chamber and is allowed to flow down the paper . The liquid moves down by capillary action as well as by the gravitational . 8

9. Contd … 4 . Circular/ Radial Development- Spot is kept at the centre of a circular paper. The solvent flows through a wick at the centre & spreads in all directions uniformly. 5. Two Dimentional Development- In this method the paper is developed in one direction and after development, the paper is developed in the second direction allowing more compounds to be separated into individual spots. 9 3 . Ascending Descending Development- A hybrid of above two technique is called ascending-descending chromatography. Only length of separation increased, first ascending takes place followed by descending.

10. APPLICATION Separation of mixtures of drugs Separation of carbohydrates, vitamins, antibiotics, proteins, etc. Identification of drugs Identification of impurities Analysis of Metabolites of drugs in blood , urine. 10

11. Contd … ADVANTAGES Simple ,rapid ,inexpensive ,excellent resolving power . Paper chromatography requires very less quantitative material. Both unknown inorganic as well as organic compounds can be identified. DISADVANTAGES Less accurate compared to HPLC or HPTLC. Complex mixture cannot be separated. Large quantity of sample cannot be applied.   11

12. Reference Vidya.Sagar .G ,2009, Instrumental Methods Of Drug Analysis. (Introduction And Paper Chromatography ),. By Publisher Pharma Med Press PP :69-71. Upadhya.A , Upadhya.K,Nath.N,2007, BiophysicalChemistry (Principle and Technique),4th Edition, Himalaya Publishing House ,PP.34. Wilson.K , Walker.M.J,1994, Electrophoretic techniques In: Principles and Techniques of Biochemistry,7th Edition, Combridge University Press, pp.425-442. 12

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