COLUMN AND THIN LAYER CHROMATOGRAPHY
Michael Dominique B. Allag, Eryll Joy H. Agojo, Camille A.Baetiong, Greanne P. Danica Ballesta,
Anne S. Barlao
Group 1 2C Medical Technology Organic Chemistry Laboratory
Chromatography is a technique for separating mixtures into their components in order to analyze,identify, purify, and quantify the mixture or components. There are different types of chromatographyand each has its own advantages and disadvantages. In this experiment, DCM-hexane was used toextract the different pigments of the siling labuyo.
Extract was introduced into the column and eluatewas collected, this process is the column chromatography (CC) method. The purity of the componentswas determined by using thin later chromatography (TLC). UV lamp was used to visualize thedeveloped TLC plate and the Retention or Retardation Factor was measured.
Chromatography can be defined as alaboratory technique that separatescomponents within a mixture by using thedifferential affinities of the components for amobile medium and for a stationary mediumthrough which they pass. The underlying principleof chromatography is that different substanceshave different partition coefficients between thestationary and mobile phases. A compound thatinteracts weakly with the stationary phase willspend most of its time in the mobile phase andmove rapidly through the chromatographicsystem. Compounds that interact strongly withthe stationary phase will move slowly. All formsof chromatography work on the same principle.Diverse types of Chromatography arepossible, depending on the physical states of the phases. Employing a gas the mobilephaseis termed gas chromatography (gc) or vaporphase chromatography (vpc). Separations usinggas chromatography involve vapor phase versusadsorption and/or equilibria. LiquidChromatography(lc)referstoanychromatographic process that employs a mobileliquid phase. Chromatographic separations canalso be carried out using thin layerchromatography (tlc) and column chromatographywhich a variety of supports, including immobilizedsilica on glass plates.Chromatography separates a substanceinto its component parts, which is very useful, assubstances are often unique in their composition.It can identify a substance and show how itdiffers from others that may look alike on thesurface. All types of chromatography areuseful for analytical purposes. Underappropriate
, all types
of chromatography can beused forpreparative scale separations. In everytype of chromatography there are threeelements to be considered: the size of thesample (Load), relative separation of components (Resolution), and the Speed.It would be
ideal ifall three elements could bemaximized so that complete separation of samples of any desired size could be quicklyachieved. In practice, generally two of theseelements can be maximized at the expense of the third. For routine analytical work,resolution and speed are maximized at theexpense of the load. In preparative scaleseparations,load, and
speed can bemaximized but then separations are usuallyincomplete. Complete separations of largesamples can be achieved but the overalloperation is likely to be slow and tedious, andmay involve the use of large quantities of solvent that must be distilled for reuse, ordiscarded.