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Procedure1. Physical State, Color and Odor The physical states must observe at room temperature.

The color and odor must also be noted. 2. Solubility Properties The sample will be introduced in a clean and dry test tube.

Experiment No. 5 Column and Thin Layer Chromatography

Liquid 4 drops of each sample will be added Solid 0.1 g of sample will be added

Extract the pigments of the red siling labuyo using DCM-hexane (1:1) and the malunggay pigments using hexane: acetone (7-3). Set aside a portion of the extracts for TLC.

Visualize the components using a UV lamp. Measure the Rf values. Document the chromatoplates.

The solid samples must be grinded to increase the surface area. The solvent was added drop wise and counted the number of drops of solvent added to a total of 3mL.

Plug the column with cotton and uniformly pack with silica gel up to the indented part of the Pasteur pipette. Place 0.5mL of the extract on top of the column using a Pasteur pipette.

Carefully place the plates in the developing chamber (see Figure 5a). Allow the solvent system to rise up to 1cm from the upper end. Remove the plates from the chamber, immediately mark the solvent front, and airdry.

Use a dry and calibrated dropper (X number of drops = 1mL). Do not heat the mixture.

3. Reaction with Litmus Paper If the sample is soluble in water, test its aqueous solution with red and blue litmus papers. Note down the color changes in both litmus papers. 4. Ignition Test -3-5 drops of the liquid sample and a pinch amount if solid will be placed in an evaporating dish and will be lighted using a match.
Note the color of the flame produced If flammable observe burning time

Discard the colorless eluate. Collect the colored eluates in separate vials. Note the number of drops of eluate collected in each vial. Apply the eluates on a 5 cm x 8 cm precoated TLC plate by spotting 10 times. Allow each spot to dry before applying the next. Make sure the spots are as small as possible.

Prepare the developing chamber by placing an approximate amount of the solvent system: DCM- hexane (1:1) for red siling labuyo and hexane- acetone (7:3) for malunggay leaves. Line the inner wall of the chamber with filter paper, cover with a watch glass, and allow equilibrating.

Spotting Of Chromatography plate

5. Infrared (IR) Analysis

Used the appendix notes to identify the type of principal bond present in the sample compound that may be observed in the IR spectra and its wave number range.

Elute the pigment mixture using 10mL of DCM- hexane (1:1) for red siling labuyo and hexane- acetone (7:3) for malunggay leaves. Introduce the solvent system in portions. Do not allow the column to run dry.