PAPER CHROMATOGRAPHY

paper chromatography

Separation of amino acid on the basis of the difference in solubility of amino acid between 2 immiscible solvents - is an analytical method used to separate colored chemicals or substances

stationary, mobile

2 phases paper chromatography

stationary phase

a solid, or a liquid supported on a solid)

mobile phase

(a liquid or gas)

liquid, gas, ion exchange, affinity

4 types

mobile/carrier

solvent moving through the column - the medium that accompanies the analyzed substance as it moves through the stationary phase

stationary

substance that stays fixed inside the column - medium on which the separation occurs

eluent

fluid entering the column

eluate

fluid exiting the column (that is collected in flasks)

elution

the process of washing out a compound through a column using a suitable solvent

analyte

mixture whose individual components have to be separated and analyzed

capillary action

Phenomenon where the solution (sometimes called the eluting solvent) will begin to rise up the paper

filter paper

stationary phase is the

developing soln

mobile phase is the

retention factor/retardation factor

This is the ratio of the distance traveled by the substance to the distance traveled by the solvent.

4:1:5 mixture of butanol, acetic acid and distilled water.

developing soln ratio

whatmen

what kind of filter paper used

0.5% of glysine, lysine, aspartic acid, unknown

amino acid used

0.2% ninhydrin solution

what did u use to spray

capillary action

solvents moves via

a. a amino acid

b. proline

a. give a blue-purple product.

b. gives a yellow orange product.

t

t/f: higher the affinity of the amino acid for the mobile phase and the faster its migration

aa distance/solvent distance

rf val formula

0

If the substance is strongly attracted to the stationary phase and not attracted to the mobile phase, then the substance does not move at all, so, the Rf is

1.0

If the substance has no affinity to the stationary phase, then the substance moves along the solvent front, so, the Rf is

Solubility of the substance to the mobile phase and Affinity of the substance to the stationary phase

the movement of the substance in relation to the solvent front (Rf) is dependent upon:2