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  • In this lesson, we will learn about paper and thin layer chromatography, both of which

  • are used in forensic science applications.

  • Chromatography is a separation technique, and this will be demonstrated using food colouring.

  • Place this in a beaker with water, ensuring that the water level does not touch the spots.

  • As the water is absorbed, it separates the multiple components of the food colouring.

  • We have now created a chromatogram.

  • The chromatography paper is called the stationary phase as itstays put”.

  • Water is called the mobile phase as itmovesup the chromatography paper.

  • Here is the key idea – a component that is more soluble in water travels a greater

  • distance up

  • the chromatography paper, and a less soluble component travels a shorter distance.

  • The retention factor, or Rf value,value, is a measure of the distance that a separated

  • component has

  • travelled compared to the distance that the solvent, in our example, water, has travelled.

  • Rf = distance travelled by a component / distance travelled by the solvent

  • These distances can be measured using a ruler.

  • Rf = distance travelled by a component / distance travelled by the solvent

  • Distance travelled by solvent = 6cm

  • Distance travelled by component A = 4cm

  • Distance travelled by component B = 2cm

  • Rf for component A = 4cm / 6cm = 0.67

  • Rf for component A = 2cm / 6cm = 0.33

  • Challenge:

  • What does the Rf value tell you about the solubility of a separated component in the

  • mobile phase solvent?

  • Please pause the video to think about this.

  • A larger Rf value means that the component is more soluble in the solvent.

  • A smaller Rf value means that the component is less soluble in the solvent.

  • Thin layer chromatography, or TLC, works in exactly the same manner.

  • Instead of chromatography paper, a glass plate with a thin layer of silica gel or any other

  • adsorbent material is used.

  • This thin layer of adsorbent material is the stationary phase.

  • Any suitable solvent may be used as the mobile phase.

  • If the separated components are colourless, the dried chromatogram may be sprayed with

  • ninhydrin, which can turn some components purple, or the chromatogram may be viewed

  • under UV light.

  • Ninhydrin is be used to detect fingerprints from a crime scene.

  • Both chromatography methods can be used to identify and compare samples of drugs, inks,

  • lipsticks, saliva, and explosives, just to name a few.

  • In conclusion, paper and thin layer chromatography are used to separate components of a

  • mixture based on their solubility.

  • The relative solubility of separated components can be determined by calculating their Rf

  • values.

  • More soluble components have a larger Rf value and less soluble components have a smaller

  • Rf value.

In this lesson, we will learn about paper and thin layer chromatography, both of which

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