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7.7 - Separation of a Mixture of Indicators

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Experiment Summary

This experiment involves separating a mixture of indicators using chromatographic techniques such as paper chromatography, thin layer chromatography (TLC), or column chromatography.

These techniques exploit the differences in the affinities of the components for a mobile phase and a stationary phase.

The different components of the mixture are identified based on their retention factors (Rf values) or their reactions in acidic and basic conditions.

Materials and Apparatus Required

Chemicals

  • Universal Indicator solution (or other mixture of indicators)
  • Methyl orange solution
  • Bromothymol blue solution
  • Phenolphthalein solution
  • Ethanol
  • Ammonia solution (0.88)
  • Water/ethanol/ammonia solution (5:2:1)

Apparatus

  • Paper chromatography tank
  • Chromatography paper
  • Capillary tubes
  • Solvent trough
  • Hair dryer

Safety Precautions

  • Wear safety glasses throughout the experiment.
  • Ethanol is flammable; keep away from open flames.
  • Ammonia solution is toxic and irritating to the lungs. Handle with care and ensure the experiment is conducted in a well-ventilated area.

Method

Paper Chromatography

  1. Prepare a chromatography tank by adding solvent (water/ethanol/ammonia solution) to a depth of 10 mm at the bottom.
  2. Cover the tank and allow it to stand for a few hours to saturate with solvent vapour.
  3. Using a pencil, draw a line near the top of a sheet of chromatography paper and another line about 3 cm from the bottom.
  4. Place small spots of each indicator and the mixture of indicators on the line near the bottom using a capillary tube.
  5. Dry the spots with a hairdryer and repeat the spotting to intensify the spots.
  6. Place the chromatogram in the tank, ensuring the solvent level is below the line where the samples are spotted.
  7. Allow the solvent to rise until it reaches the top line.
  8. Remove the chromatogram, dry it, and measure the distance travelled by the solvent front and by each indicator spot.
  9. Calculate the Rf value for each indicator:
Rf=Distance travelled by the componentDistance travelled by the solvent front\text{Rf} = \frac{\text{Distance travelled by the component}}{\text{Distance travelled by the solvent front}}
  1. Identify the indicators in the mixture by comparing their Rf values with known values.

Results

IndicatorDistance Travelled (cm)Rf Value
Methyl orange6.80.68
Phenolphthalein8.40.84
Bromothymol blue9.70.97
Fastest component of mixture9.70.97
Second component of mixture8.40.84
Third component of mixture7.90.79
Fourth component of mixture6.80.68

Indicators present in the mixture: Bromothymol blue, methyl orange, phenolphthalein, and at least one other.

Example Questions with Answers

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Q1: Why is a chromatography tank allowed to stand for a few hours after the solvent is added?

The tank needs time to saturate with solvent vapour, ensuring optimum and reproducible results.

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Q2: What is the significance of the two lines drawn on the chromatography paper?

The lower line indicates the starting position of the sample spots, while the upper line marks the distance travelled by the solvent front, which is used to calculate Rf values.

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Q3: What do different Rf values indicate in paper chromatography?

Different Rf values indicate that the components of the mixture are not identical, as they have different affinities for the mobile and stationary phases.

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Q4: How can the components of a mixture be identified using chromatography?

The components can be identified by comparing their Rf values with the known Rf values of standard indicators under the same experimental conditions.

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