University:
Swansea University
Course:
PM134 | Microbiology
Academic year:

2023
Views:

122

Pages:

2
Author:

Sanjeev K

ERIMENT NO DM106-10-2022 THINLAYER CHROMATOGRAPHY AIM Seperation and identification of •of plam pigments wing thin layer chowmatography technique PRINCIPLE Thin layer chomatography inolva flowing f is a planar chromatography, Omobile phase over a thin laybu adsorbent applied on sotid 'comperents exeur by st, where suppost eparation differential migration which Beers when solvent flores along fine powder spread on glass plates Here in Rayered on Malubnary phase is silica or on a The plate and mobile phase is Retardation fautos; alumina liquid. R₁ = Distance travelled by the solute Distance travelled bitne solvent by the MATERIALS REQUIRED The plate, beaker, capillary tube, Palak leaves, mortar and Pestle, retone, water, mobile phase (Petroleum ethur: isopropa nol: Weder), Samples, Peruil, seale, aluminium foru PROCEDURE *~1 ~Igm of Palak leaves are taken in a mortar, in which * Equal parts avetone and water are added. Using a pestle, the leaves are ground or homogenized to a liquid sample. * The plate f a particular dimension is taken. * An origin line is drawn from the bottom end & using smale and penul, Ikm the TLCO Plate. * Two points are marked with equal distanus for spolling two samples; where one is to spor a known sample B-carotene 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 it is allowed to dry The The plate is then dipped in the bolvent (Petroleum aner: isopnpand; water - 10:1:0.025) taken in a beaker The beaker is wrapped using on the mouin part to an aluminium fois andill evaporation * The Lolvent must always be below the muns origin lime 30-45 *fe approximately takes to rise through the The plate for the solvent by capillary action. * Once the solvent ent front is developed, the the Plate is removed, immediately marked with a penul and It is allowed dry * To enable drying, as well as to get quick results the TLC plate is kept is hot air oven and it is observed. * Omeen coloured chromatogram is obtained * Using seale the distance travelled by the solvent from and the distance travelled by the solute is noted. *Then Re value is calculated and compared with the standard referance table to get the results RESULTS Re value of unknown sample =0.5 Day's Re value is close to the Re value of Chlorophyll bas bas given in the standard siferance table. . The unknown sample is chlorophyll b