paper chromatography.pptx principle and applications

Paper chromatography principle, practical requirrments, Rf value and applicatipns.

1. PAPER CHROMATOGRAPHY PRESENTED BY :- M. UMA SRI LAKSHMI 22PE1T0013 IV PHARM D

2. CONTENTS INTRODUCTION 1 PRINCIPLE OF SEPARATION 2 PRACTICAL REQUIREMENTS 3 Rf VALUE 4 APPLICATIONS 5

3. INTRODUCTION:- Paper chromatography is defined as a technique in which the analysis of unkown substance is carried out mainly by the flow of slovents are designed filter paper .

4. Two types :- PAPER ADSORPTION CHROMATOGRAPHY :- In which the paper impregnated with silica or alumina action as adosrbent (stationay phase) and solvent as mobile phase. PAPER PARTITION CHROmATOGRAPHY:- in which the water present in the pores of cellulose fibres present in filter paper acts as stationary phase and another mobile phase is used as solvent.

5. PRINCIPLE OF SEPARATION:- The principle of separation is mainly partition rather than Adsorption cellulose layers in filter paper contaions moisture which action as stationary phase. Organic solvents or buffers are used as mobile phase. Instead of water As stationary phase, other organic solvents can be used by suitable modification.

6. PRACTICAL REQUIREMENTS STATIONARY PHASE AND PAPERS USED APPLICATION OF SAMPLE MOBILE PHASE D evelopment technique Detecting or visualizing agents

7. 1. STATIONARY PHASE AND PAPER USED Paper of chromatographic grade consists of alpha- cellulose-98 .999 Beta- cellulose – 0.3-1%, pentosans-0 .4-0.8%, ethens slouble matter-0.015-15 . whattsman filter paper of different grade like size,shape and thickness. Choice of filter paper depending upon thickness, flowrate, purity, technique etc. MODIFIED PAPERS:- ACID OR BASE WASHED FILTER PAPER, GLASS FIBRE TYPE PAPER. HYDROPHILLIC PAPERS :- PAPERS MODIFIED WITH METHANOL, GLYCOL, GLYCEROL ETC. HYDROPHOBIC PAPER:- ACETYLATION OF ‘OH’ GROUP LEADS TO HYDRPHOBIC NATURE HENCE CAN BE USED FOR REVERSE PHASE CHROMATOGRAPHY. SIZE OF THE PAPER USED : - PAPER OF ANY SIZE CAN BE USED PAPER SHOULD BE KEPT IN A CHAMBER OF SUIATBLE SIZE.

8. 2.APPLICATION OF SAMPLE :- The sample to be applied is dissolved in the mobile PHASE and applied using capillary tube or using micropipette. 3. MOBILE PHASE Pure solvents, buffer solution or mixture of solvents are used. Ex:- HYDROPHILLIC MOBILE PHASES. Isopropanol: ammonia : water-9 :1:2 Butanol : glacial acetic acid : water – 4:1:5 Methanol : water – 3:1 / 4:1

9. 4. DEVELOPMENT TECHNIQUE:- Since paper is flexible when compared to glassplate using TLC, several types of development are possible which increase the ease and efficiancy of operation. They are ASCENDING DEVELOPMENT DESCENDING DEVELOPMENT ASCENDING AND DESCENDING DEVELOPMENT CIRCULAR OR RADIAL DEVELOPMENT TWO DIMENSIONAL DEVELOPMENT

10. 5. DETECTING OR VISUALING AGENTS :- NON SPECIFIC METHOD :- • Iodine chamber method • UV chamber for flouroscent compounds SPECIFIC METHOD :- • Destructive technique • non destructive technique. After the development of chromatogram the sports should be visualized. Detecting coloured sports can be done usually.

11. Rf VALUE :- The rf value is calcualted for identifying the spots THAT IS IN QUALITATIVE ANALYSIS. RF VALUE IS THE RATIO OF DISTAJCE TRVELLED BY THE SOLUTE TO THE DISTANCE TRAVELLED BY THE SOLVENT FRONT. RF VALUE = DISTANCE TRAVELLED BY SOLUTE DISTANCE TRAVELLED BY SOLVENT Front . Rm VALUE :- Rm value is used in qualitative analysis to find out weather the compounds belongs to a homologous series. Rm=log [1/rf-1]

12. APPLICATIONS :- Paper chromatography is more useful for the analysis of polar compounds like amino acids , sudarsn and natural products ETC. Separation of mixtures of drugs of chemicals or biological origin, plant extracts ETC. Separation of carbohydrates , vitamins , alkaloids, glycosides, amino acid ETC. I dentification of foreign sibstances in drugs. Analysis of metabolites of drugs in blood, urine ETC.

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