RP7 Chromatography of photosynthetic pigments

Free AQA A-level Biology RP7 questions on Chromatography of photosynthetic pigments, including practical reasoning, variables, data handling, clues and explanations.

Specification route
RP7
Question bank
13 questions
Course stage
Year 13 / A-level only

Sample questions

QUESTION 1 · RP7 · LEVEL 1

When setting up paper chromatography to separate photosynthetic pigments, why must the line at the origin be drawn in pencil rather than ink pen?

  • Ink contains soluble pigments that would dissolve in the solvent and travel up the paper, obscuring the results.
  • Pencil graphite acts as a catalyst for the separation of the plant pigments.
  • Ink reacts chemically with the chloroplast extract, destroying the chlorophyll.
  • Pencil lead is hydrophobic, preventing the solvent from rising too quickly.
Show clue

Think about what the solvent is meant to dissolve and carry.

Show answer and explanation

Answer: Ink contains soluble pigments that would dissolve in the solvent and travel up the paper, obscuring the results.

Required practical 7. Pencil graphite is insoluble in the chromatography solvent. Pen ink contains dyes that would dissolve and run up the chromatogram, interfering with the plant pigments.

QUESTION 2 · RP7 · LEVEL 2

When setting up paper chromatography, it is essential that the solvent level in the beaker sits below the pencil origin line. Why?

  • If the solvent covers the origin line, the concentrated plant pigments will dissolve directly into the solvent pool instead of travelling up the paper.
  • If the solvent is too high, it will wash the pencil graphite off the paper and obscure the results.
  • The paper will become too heavy with solvent and tear under its own weight.
  • The solvent will evaporate before it can carry the pigments up the chromatogram.
Show clue

The pigments must be drawn upwards by capillary action, not washed away.

Show answer and explanation

Answer: If the solvent covers the origin line, the concentrated plant pigments will dissolve directly into the solvent pool instead of travelling up the paper.

Required practical 7. The pigment spot must sit above the solvent. If submerged, the pigments will simply wash off the paper into the bulk liquid at the bottom of the beaker.

QUESTION 3 · RP7 · LEVEL 2

Why must the position of the solvent front be marked with a pencil immediately upon removing the chromatogram from the beaker?

  • Because the highly volatile solvent will evaporate very quickly, making the solvent front invisible and preventing the calculation of the $R_f$ value.
  • Because the plant pigments will continue to travel up the dry paper until they hit the pencil line.
  • To stop the solvent from chemically reacting with the oxygen in the air.
  • Because the pencil graphite acts as a chemical fixative to permanently set the pigment colours.
Show clue

The distance the solvent travels is the denominator in the $R_f$ calculation.

Show answer and explanation

Answer: Because the highly volatile solvent will evaporate very quickly, making the solvent front invisible and preventing the calculation of the $R_f$ value.

Required practical 7. Chromatography solvents (like propanone or petroleum ether) are highly volatile. If the front isn't marked immediately, it evaporates, making it impossible to measure the distance the solvent moved.

QUESTION 4 · RP7 · LEVEL 2

When spotting the plant extract onto the chromatography paper, why must the student allow the drop to dry before adding another drop exactly on top of it?

  • To build up a highly concentrated spot of pigment without allowing the spot to spread out too widely, which would blur the final separated bands.
  • To chemically lock the pigments into the cellulose fibres of the paper.
  • To evaporate all the water from the extract so only the pure chloroplasts remain.
  • To prevent the pencil origin line from dissolving into the extract.
Show clue

You want a tiny, dark dot. If you keep adding liquid without drying, the puddle just gets wider and wider.

Show answer and explanation

Answer: To build up a highly concentrated spot of pigment without allowing the spot to spread out too widely, which would blur the final separated bands.

Required practical 7. The pigment spot must be as small and concentrated as possible. Drying between applications prevents the drop from spreading out, which ensures the separated pigment bands will be sharp and distinct rather than blurred together.

QUESTION 5 · RP7 · LEVEL 2

Why is the chromatography beaker or tank tightly covered with a lid during the experiment?

  • To saturate the air inside the tank with solvent vapour, preventing the solvent from evaporating off the surface of the paper as it rises.
  • To create an anaerobic environment, preventing the oxidation of the delicate photosynthetic pigments.
  • To block out light, stopping the light-dependent reaction from destroying the chlorophyll.
  • To increase the atmospheric pressure inside the tank, forcing the solvent up the paper faster.
Show clue

The solvents used (like propanone or petroleum ether) are highly volatile and turn into gas very easily at room temperature.

Show answer and explanation

Answer: To saturate the air inside the tank with solvent vapour, preventing the solvent from evaporating off the surface of the paper as it rises.

Required practical 7. Chromatography solvents are highly volatile. Covering the tank prevents the solvent from evaporating from the paper before it reaches the top, ensuring the pigments are carried the maximum possible distance.

QUESTION 6 · RP7 · LEVEL 3

What determines how far a specific photosynthetic pigment travels up the chromatography paper?

  • Its relative solubility in the mobile phase (the solvent) compared to its affinity for the stationary phase (the paper).
  • Only its physical molecular mass; lighter pigments always travel the furthest.
  • Its specific colour; darker pigments are heavier and stay near the bottom.
  • The rate at which it absorbs light energy during the experiment.
Show clue

Chromatography separates mixtures based on how much the molecules 'like' the liquid moving them versus the solid they are moving across.

Show answer and explanation

Answer: Its relative solubility in the mobile phase (the solvent) compared to its affinity for the stationary phase (the paper).

Required practical 7. Separation in chromatography is based on differing solubilities and affinities. A pigment highly soluble in the solvent and with low affinity for the paper will travel further up the chromatogram.

QUESTION 7 · RP7 · LEVEL 2

A student calculates the $R_f$ value of a pigment to be $1.2$. What is the most likely explanation for this result?

  • A calculation error has occurred; the $R_f$ value cannot be greater than $1.0$ because the pigment cannot travel further than the solvent front.
  • The pigment is highly soluble in the solvent and has zero affinity for the paper.
  • The experiment was run for too long, allowing the pigment to detach from the paper.
  • The student used the wrong solvent, causing the pigment to be repelled upwards.
Show clue

Look at the formula: $R_f$ = distance moved by spot / distance moved by solvent. The numerator can never exceed the denominator.

Show answer and explanation

Answer: A calculation error has occurred; the $R_f$ value cannot be greater than $1.0$ because the pigment cannot travel further than the solvent front.

Required practical 7 / MS 0.3. The $R_f$ value is a ratio. Because the solvent front is the absolute maximum distance anything can travel, the distance moved by the pigment can never be larger than the distance moved by the solvent.

QUESTION 8 · RP7 · LEVEL 1

What is the primary safety hazard associated with the solvents (e.g., propanone, petroleum ether) used in plant pigment chromatography, and what is the key control measure?

  • Hazard: Highly flammable. Control Measure: Ensure there are no naked flames (like Bunsen burners) in the laboratory.
  • Hazard: Highly radioactive. Control Measure: Store in lead-lined containers.
  • Hazard: Corrosive acid. Control Measure: Neutralise with a strong alkali before use.
  • Hazard: Contains live pathogens. Control Measure: Autoclave the solvents before use.
Show clue

These liquids vaporize easily and catch fire very quickly.

Show answer and explanation

Answer: Hazard: Highly flammable. Control Measure: Ensure there are no naked flames (like Bunsen burners) in the laboratory.

CPAC 3: Assess risks. The organic solvents used to extract and separate plant pigments are highly flammable. The primary safety precaution is keeping them strictly away from naked flames.

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