Paper chromatography-Introduction, methodology, development techniques, advantages, disadvantages and applications
Electrophoresis– Introduction, factors affecting electrophoretic mobility, Techniques of paper, gel, capillary electrophoresis, applications
3. Pneparation of Paper:
Once the type of paper is decided, it is cut in desired size and shape depending upon the work to be carried out. Generally rectangular and square shapes are used. After noting the direction of run on paper, startline is marked as shown in Fig. 5.1. While storing, paper should be kept away from any fume (especially ammonia) and should not be subjected to large changes in humidity.
4. Preparation of sample:
The mixture to be separated is applied to the paper as a solution. It is important to choose proper solvent for making solution. Generally, a weighed amount of mixture is dissolved in volatile solvent and a minimum volume of concentrated solution is applied on the paper avoiding diffusion of spot. Extracts from soil, biological cells or tissue materials are taken out with the help of some solvent which is directly applied on the paper.
Aqueous biological extracts, urine, neutralised protein hydrolystates and other solutions which may have to be examined for amino acids and sugars will always contain appreciable
amounts of inorganic material. Removal of these is called “desalting” and should always be carried out without affecting organic compounds. Desalting is carried out by electrolytic method, electrodialysis, ion-exchange membrane, column techniques etc.
No standard procedure for preparation of samples could be presented here, as several factorsaffect. Commonly, sample volumes of 10-20 p/ containing as many p,g of substance is spotted,
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