computer application bioinstument and biostatistics Neha yadav.pptx

computer application bioinstument and biostatistics Neha yadav.pptx

1. Topic–paper chromatography Session :2022-2023 Paper- Computer application bioinstument and biostatistics Course Code : BTC S01 course type: OSC Directed by presented by Mr. Avdhesh kumar Neha yadav class-M.sc 1 st semester (biotechnology )

2. Content Introduction Principle of Paper Chromatography Phases of Paper chromatography Types of Paper chromatography Paper chromatography Procedure Rf values Application of Paper chromatography Advantages of Paper Chromatograph Limitations of Paper Chromatography Reference

3. Paper chromatography (PC) is a type of planar  chromatography  whereby chromatography procedures are run on a specialized 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. It was first introduced by German scientist Christian Friedrich Schonbein (1865). What is Paper Chromatography?

4. Principle of Paper Chromatography 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 .

5. Phase of Paper chromatograp 1. mobile phase :   mobile phase is combination of non-polar organic solvents. The solvent runs up the stationary phase via capillary movement. 2. stationary phase : stationary phase is polar inorganic solvent, i.e. water. Here, the absorbent paper supports the stationary phase, i.e. water. ☆ papers used Whatman filter papers of different grades like No.1, No.2, No.3, No.4, No.20, No.40, No.42 etc In general the paper contains 98-99% of α- cellulose, 0.3 – 1% β - cellulose.

6. Types of Paper chromatography • Two types of paper chromatography Paper adsorptions chromatography : In the clinical laboratory it is used to employed to detect and identify sugar and amino acids . Solid + liquid Paper partition chromatography : The principal involved is partition chromatography where in the subtances are distributed or partitioned between liquid phases.one phase is the water which is held in the pores of the filter paper used and other one is mobile phase which is moves over the paper . liquid + liquid

7. Paper Chromatography procedure Selection of Solid Support Selection of Mobile Phase Saturation of Tank Sample Preparation and Loading 5 . Development of the Chromatogram Drying of Chromatogram Detection

8. Rf values Light green spot indicates chlorophyll-b pigment. = 2/10 = 0.2

9. Methods of Paper chromatography

10. Applications of Paper chromatography • Separation of Ink Dyes -To compare ink dyes use in any company • Food Colouring - To differentiate colouring agent used in food products • To isolate plant pigment from root and leaves • Separation of mixture of drugs • Separation of carbohydrates , vitamins , amino acids etc.

11. Advantages of Paper Chromatograph • Simple • Rapid • Paper Chromatography requires very less quantitative material. • Paper Chromatography is cheaper compared to other chromatography methods. • Both unknown inorganic as well as organic compounds can be identified by paper chromatography method. • Paper chromatography does not occupy much space compared to other analytical methods or equipments. • Excellent resolving power

12. Limitations of Paper Chromatography • Paper chromatography cannot handle large amounts of sample. • Paper chromatography is ineffective in quantitative analysis. • Paper chromatography cannot separate complex mixtures. • Less Accurate than HPLC or HPTLC

13. 2 .https://laboratoryinfo.com/paper-chromatography/ REFERENCE 1 . Ahuja satinder, chromatography and separation science , Volume 4 , Academic press 2002 , chepter- 7 paper chromatography , page no . – 102 - 111 3 . https://microbenotes.com/paper-chromatography/