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Chloroplast pigments chromatography CPAC

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Full CPAC write up which achieved passing grade and fulfilled all necessary criteria for Biology Edexcel for chromatography practical looking at pigments in chloroplast

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February 12, 2023
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Name: Hiba Majid 13E 08/09/2022 (P:\Science\Mr Alim\Year 12\AS Biology)



QUESTION TO BE INVESTIGATED: What different chloroplast pigments are present in spinach ?

SCIENTIFIC RESEARCH (REFERENCED):

● Pigments are coloured compounds, and are involved in the dependent reaction in
photosynthesis. they are extracted from the thylakoid membranes of the chloroplast
● Pigments are insoluble but as finely ground particles mixed with a liquid
● Acetone is able to dissolve both polar and nonpolar substances (Degreasers, 2022)
● This process is important where “In order to extract these pigments from the thylakoid
membranes of the chloroplasts, the organelles in which photosynthesis occurs, fresh,
ground or torn leaves (preferably spinach) may be soaked in acetone” (Separation of
Pigments by Paper Chromatography, 2022)
● Spinach has green chlorophylls and the yellow, orange, and red carotenoids. Chlorophyll a
levels are higher than b
● Chromatography is a process for separating components of a mixture. To get the process
started, the mixture is dissolved in a substance called the mobile phase, which carries it
through the stationary phase. The different pigments travel at different speeds, causing
them to separate. (Staff, 2022)

Risk assessment:

Hazard Risk Protective What to do Risk level
measures

Propanone Flammable; Use eye Wash hand with Low
causes eyes protection, make cool water, seek
irritation; sure the room is medical
breathing may well-ventilated, assistance
result in and keep solution
lightheadedness away from open
and drowsiness, flames.


Chromatography Flammable; Use eye Wash hand with Low
solvent irritating to the protection, make cool water, seek
eyes and skin; sure the room is medical
dangerous when well-ventilated, assistance
inhaled and keep solution
away from open
flames.

Biohazard Bacteria may lead Wash hands after Seek medical Low
to infections, use assistance
contamination




Mr Alim

, Name: Hiba Majid 13E 08/09/2022 (P:\Science\Mr Alim\Year 12\AS Biology)



METHOD

1. Carry out instructions given (or those you have produced and researched) to complete the
practical (CPAC1)
2. List apparatus below and describe techniques used when measuring quantities with a
particular piece of apparatus:

Piece of apparatus Purpose of piece of Comment on how piece of equipment Appendix
apparatus was used 5c
accurately/appropriately/safely technique

Spinach leaves Obtain pigments from its Hands were washed thoroughly after
chloroplast touching to avoid contamination and
bacteria

Low risk

Weighing scale Weigh out mass of We measured out 5 grams as this was
spinach in grams sufficient

Kept in the middle of table to avoid
falling off and breaking

Low risk

Sand To help break up the Only a pinch was used as this was
plant cells sufficient

Eye protection used to avoid going in
eye

Low risk

Pestle and mortar This was used to grind the We used the corkscrew technique to
spinach leaves in the ensure the pigments were released
propanone properly

Kept in the middle to avoid ceramic
breaking

Low risk

Gas jar To place the stationary This was kept in the middle to avoid it
and mobile phase in falling off the table

Low risk

Pencil To draw the origin line We use a pencil to draw the origin line
and draw around the because if we used a pen instead the
pigments at the end of pigments in the ink may interfere with



Mr Alim
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