Paper Chromatography

EXPERIMENT NO: 1 DATE: 29.09.2022 PAPER CHROMATOGRAPHY AIM Separation and identification of aminoarids using Paper inromatography technique. PRINCIPLE Paper enromatography is an analytical mixture technique, takes place due separation to differential of migration two immissible where components of a that occurs between the two phases (ie, partition between inromatography. mo.1 filter where paper) stationary and mobile phase components occur due liquids). It is a type phase of is paper(eg:- is liquid whatman to difference The in separation parties of co-efficient, which is related to retardation where retardation Rf factor; = Distance travelled by the solvent solute Distance travelled by the the MATERIALS no.1 filter paper (stationary phase) amino pinned REQUIRED on a solid / unknown), support mobile phase, chromatography mixed Whatman (stick), samples (3knoun jar, and penic, and scale, capillary tube, Ninhydrin with aretone (img/mi) PROCEDURE What(man no:1 filter paper of a particular dimension is taken An origin line is drawn using scale and pencil, 2cm from the bottom end of the paper. 4 points are marked with equal distances for spotting the 4 samples; where 3 known aminoarid samples and the 4th amino sample is unknown are spotted; Argining,Alamine and and Tyrosine respectively The samples are spotted and It is allowed to dry

2 The paper is then dipped in the solvent (butanol autic and: water 4:1:1) taken in the chromatography chamber The solvent must always be below the origin solvent line It takes approximately 20.30 minutes aiwon for the to rise throughthe paper by capillary is removed Once the solvent front is developed the paper allowed It is marked with a pensil and then it is to It is dry then sprayed with Ninhydrin colourless solution mixed with acetone since the samples are After spraying, for getting quick results It is kept in Purplish hot air pink oven coloured chromalogram is obtained of and It is observed different height; but the fourth unknown hence sample It is All the three samples has corresponding peaks a combination got 9 two different amino solvent has two peaks of two different heights; acid samples. and Using the distance travelled by the scale the distance travelled by solute the is noted front Then Rf value for each sample is calculated RESULTS Rf value of Argimine = 0.667 Rf value of Alamine = =0.871 0.820 Rf value of unknown sample, 1st peak = 0.564 Rf value of Tyrosine 2nd peak = 0.794 These two Rf values are close to the Rf values of Arginine and Alanine The unknown sample is a combination of amino acids, Arginine and Alamine Signa