Chemistry · Virtual lab
Thin Layer Chromatography
Separate a mixture of amino acids on a silica plate, develop it with ninhydrin, and identify the unknowns from their Rf values.
Background
Thin layer chromatography (TLC) is a rapid analytical technique for separating small quantities of mixtures. A thin layer of adsorbent (usually silica gel or alumina) is coated on a glass or plastic plate. The sample is spotted near the bottom, and the plate is placed in a developing chamber with a shallow pool of solvent.
As the solvent ascends by capillary action, components of the mixture travel at different rates depending on their polarity and affinity for the stationary phase. After development, spots are visualised using UV light, iodine vapour, or chemical reagents such as ninhydrin for amino acids.
In this virtual lab you spot a mixture of amino acids on a TLC plate, develop it in a suitable solvent system, spray ninhydrin to reveal the separated spots, and identify the unknowns by comparing their Rf values with known standards.
What you will learn
- Prepare a TLC plate and apply sample spots
- Develop the chromatogram in a solvent chamber
- Visualise separated components using ninhydrin reagent
- Calculate Rf values and identify unknown amino acids
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