Chromatography Flashcards

1
Q

What is the mobile phase

A

The solvent eg water

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2
Q

What is the stationary phase

A

Whatever the solvent is travelling up eg filter paper

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3
Q

What are the two factors that effect distance travelled by solute

A
  1. Retention by stationary phase
  2. Solubility by moving phase
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4
Q

Describe a non polar molecules movement

A
  • greater solubility in moving phase
  • less retention by stationary phase
  • so held in stationary phase for less time (will travel further)
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5
Q

Describe a polar molecules movement

A
  • less solubility in moving phase
  • greater retention by stationary phase
  • so held in stationary phase for longer (will travel less)
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6
Q

In gas chromatography what states are the mobile and stationary phases

A

Mobile - gas
Stationary - liquid

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7
Q

If the stationary phase was polar and moving phase was non polar what type of compounds would travel further/quicker

A

Non polar compounds because they have greater solubility in the moving phase

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8
Q

Why do you use a pencil line

A

Will not dissolve in solvent

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9
Q

Why do you use a capillary tube to get a tiny drop of solution

A

Too big of a drop will cause different spots to emerge

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10
Q

Why only 1cm depth of solvent

A
  • It must be below start line
    If the solvent is too deep it will dissolve the mixture from the plate
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11
Q

What do you use if the spots are colourless and not visible

A

UV lamp

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12
Q

In gas chromatography why is it important to use an inert gas

A

So it doesn’t react with the compounds being separated in the column

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13
Q

Why does gas chromatography work

A
  • different retention times
  • different balance between solubility in moving phase and retention by stationary phase
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14
Q

Why is the plate left to dry in a fume cupboard

A

Solvent is toxic

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15
Q

How to locate position of amino acids

A
  • spray with a developing agent
  • work out Rf value
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16
Q

Why do you use a lid

A

To prevent evaporation of toxic solvent

17
Q

Suggest a suitable reagent for the hydrolysis of a protein

A

Conc HCl

18
Q

Why is it necessary to use two different solvents for amino acids

A

Some amino acids didn’t dissolve with the first

19
Q

Is it essential to allow the solvent to reach all the way to the top to calculate a Rf value

A

Not essential but will make Rf value more accurate

20
Q

Why do amino acids have different Rf values

A
  • they have different polarities
  • so different retention to stationary phase and solubility in mobile phase
21
Q

What happens if your sample is too concentrated?

A

The spots will overlap