Thin Layer Chromatography

EXPI RIMINI NO:2 MIL :06.10.2022 THINLAYER CHROMATOGRAPHY AIM Separation and identification of plan pigments using thin layer annomatography technique PRINCIPLE This layer chromalography is a planar chromatography, when involves flowing of mobile phase over a thinlayer of adsorbent applied oh solid support, where separation of components occur by differential migration which Occurs when solvent flows along fine powder spread on glass places Here stationary and mobile phase is sitica is or alumina layered on a TLC plate phase liquid. Recardation failor; Rf Distance travelled by the solute Distance travelled by the solvent MATERIALS REQUIRED TLC plate, beaker, apillary tube, Palak leaves, mortar and Pestle, acctone, water, mobilephase (Petroleum ether: Isopropa not: water), samples, Penil, male, aluminium foil PROCEDURE ~ gm of Palak leaves are taken in a mortar ,in which equal portions of autone and water is added Using a pestle, the leaves are grinded or homogenized to a liquid sample TLC plate of a particular dimension is taken. An origin line is drawn using scale and pencil, 1cm from the bottom end of the TLCI plate Two points are marked with equal distances for spotting two samples; where one is to spot a known sample B-carotine and the other one is to spot the homogen- ate me homogenate is spotted 2-3 times for better results The samples are spotted and H is allowed to dry

The TLC 16 then dipped in the solveni (petroleum in a ever isopropomol: plate water 10:1 : 0.025) taken beaker is wrapped using an evaporation aluminium foil The on the beaker mouth part always to avoid be below the the origin solvent line The solvent must takes 30-45 mins for action. H approximately the The plate by capillary the TLC plate to Once removed, rise the through solvent immediately front is developed, marked with a pencil and is It is allowed to day quick results the TLC to TO enable 15 kept drying, in hot as well air as oven get and It is observed place Green coloured the enromatogram distance travelled is obtained by the solvent is noted. front using scale distance travelled by the solute with the and Rf the value is calculated and compared results standard Then referance table to get the RESULTS Ins Rf value is close to the Rf value of chlorophyll Rs value of unknown sample = 0.5 b as given in the standard referance table. The unknown sample is chiorophyll b Not