Experiment No.

Date

Aim:- To select an appropriate mobile phase for se paration of aspirin and paracetamo!.

Requirements: Grease free slide, development chamber, filter paper

Reagents: -
Silica gel GF, methanol, aspirin, paracetamol,chloroform, ethanol, hexane.

Procedure:
medium flame. Hold a
bunsen burner and adjust for a
1 Preparing a spotting capillary:- Light a

pullthe two end of the

melting point capillary in the flame until ajust begins to soften, there quickly
formed.
capillary in opposite direction. Capillary with very small diameter is

silica gel TLC plate make using pencil straight and parallel ( to
a line
2)Making the TLC plate: On a
the short dimension of the plate) to the end of the Plate about 1 cm from the end of plate. This is
This is the solvent
the line of origin. Mark another straight line parallelto the other end of the plate.
second line
front line. When you the plate you will allow the solvent front to rise to the
develop

at 100°C for 30 minutes to achieve it.

Activating
3Activation theTLC plate: Place marked TLC plate in an oven
-

involves driving off water molecule that bound to the polar site on the site.

solution of
With the help of a spotting capillary place small drop of
4 Spotting the TLC plate: to evaporate completely and
substance to be analysed. One of the 3 pencil lines. Allow the solvents
make second deposit on the same spot if desired.

mobile phase in the chamber and line

StheDeveloping the TLCplate Place the required quantityof
chamberwith filter paper. Close with the lid and allow chambe r to saturate. Place the TLC plate
the solution. The plate should be
into the beaker taking care not to immerse the spotted line into
the TLC plate (by capillary action). Allow the
kept at 45 degree angle. Allow the M.P to travel up
remove the plate from
solvent to rise till the line marked as the solvent front. After the development

Cover

Glass
Lank

TLC plate

Sorbent

Origin
Solvent

the chamber.
 6 Drying the plates: - Place the plate on flat dry surface and allow the solvent to evaporate. If
solvent is not highly volatile, place the slide on a flat surface in an oven at 50-60°c. When plate is dry
it's ready for visualization. Observe the plate under short (254 nm) and long wavelength (366nm).
Using pencil draw a boundary around each spot that matches shape of spot. Mark the centre of the
spot.

8 Measurement of Rr - The vertical distance between the line of origin to the solvent from the

measured. This is taken as the distance travelled by the solvent. The distance Iine of origin to center
of a spot is measured. This is taken as the distance travelled by the solute (analyte). Ratio of
distance travelled by the solute over the distance travelled by the solvent is the Rf or the
Retardation Factor. This value aids in the identity of the unknown analyte. (does not confirm the

identity).
Eluotropic series: The elutopic se ries lists solve nts in order of their power to carry a given compound
through stationary phase (eg. silica gel). The order of solvents runs in the order of their polarity. The
series is useful when selecting a solvent for a particular separation where more polar compounds
will normally require a more polar solvent.

High Polarity
Water
AcetiCAddiEthanoicAcid) 9
Methanol

Ethanol ..

Propan-1-ol
AcetonitrileEthanenitrile)7 A
Ethyl Acetate (Ethyl Ethanoate)

AcetonePropanone)
Dichloromethane
Chloroform
Diethyl Ether
Toluene(MethylBenzene)
Cyclohexane
D-Hexenes
LowPolarity
ON THE SAME PLATE
FOR M.P DEVELOPMENT TAKE TLC RUNS OF THE MIXTURE AND STANDARD
RUNS IN NEAT
IN NEAT sOLVENTS ( SOLVENTS WITH HIGH AND LOW ELTUING STRENGHS).
sOLVENT WILL ACT AS A REFERENCE FOR ADJUSTING THE POLARITY OF THE SOLVENT/STRENGTH
IN CERTAIN RATIO) SO
OF THE SOLVENTS TO BY MIXING SOLVENTS OF DIFFERENT STREINGTHS
(
AS TO GET A SOLVENT THAT GiVES GOOD SEPARATION OF THE COMPONENTS OF A MIXTURE.

REPORT: