liquid liquid

LIQUID-LIQUID PARTITIONING

Non-polar solvent

Aqueous solution

Separates by molecules of differing solubilities based on their polarity in two solvents that won't mix (chloroform and water)

Prepare the Mixture: Start with a mixture containing the molecules you want to separate. If the mixture is in a water-based

solution, you'll add chloroform to

create a two-phase system.

01

Co

02 Moxing and Phase Separation: Add chloroform to the

mixture and vigorously shake or stir the solution. Over time, the mixture will separate into two distinct phases due to the immiscibility of water and chloroform. The more polar molecules will tend to stay in the water phase, while less polar molecules will preferentially partition into the chloroform phase.

Separation: Allow the mixture to settle so that the two phases fully separate. The denser chloroform phase will typically be at the bottom, and the less dense water phase will be on top.

90

04

03

Collection: Carefully remove the chloroform phase from the bottom using a separating funnel or a similar apparatus. This phase will contain the less polar molecules.

Recovery: The aqueous (water) phase remaining on top will contain the more polar molecules. This phase can also be collected if needed.

05

This separation technique exploits the fact that polar molecules have a stronger affinity for water due to their ability to form hydrogen bonds, while non-polar tend to prefer the organic solvent (like chloroform). By using these immiscible solvents, you can effectively partition

molecules based on their relative polarity.