Analytical techniques - chromotography

Question 1

chromatography

Answer 1

used for separating a molecule into compounds

Question 2

mobile phase

Answer 2

moves over the stationary phase and carries components of sample with in

Question 3

stationary phase

Answer 3

stays still + has an origin where sample starts. Components will move with the mobile phase based on how strongly attracted they are to the mobile or stationary phase (depends on IMF)

Question 4

paper chromatography

Answer 4

components are more attracted to the mobile phase (more similar IMF) will travel further along the stationary phase

Question 5

collum chromatography

Answer 5

• solid stationary phase is resin/gel beads packed into a column
• the sample is placed at the start of the column
• mobile phase (solvent) is passed through the column and through the tap at the bottom
• components of the sample are more/less attracted to one of the phases and pass through the column at different rates

Question 6

Size exclusion

Answer 6

• The stationary phase has pores that catch smaller proteins and larger proteins leave the column first

Question 7

ion exchange

Answer 7

• Components separated based on charge
• column has positive or negative beads
• proteins have charges based on R groups, chains and amino/carboxyl groups

Question 8

hydrophobic interactions

Answer 8

• hydrophobic proteins have higher affintity with stationary phase and will take longer to pass through

Question 9

affinity

Answer 9

resin is coated in a ligand (protein) which will bind with a specific protein of interest. Once sample has moved through column, resin is flushed with a chemical to detach protein

Question 10

high-performance liquid chromatography

Answer 10

• type of column chromatography
• a detector at the bottom of the column detects components passing through and produces a peak
• time taken for a component to pass through (retention time) similar to retardation factors

Question 11

separating proteins in a sample (size) - electrophoresis

Answer 11

• uses gel as a stationary phase and passes a current through (negative and positive ends)
• The protein sample is coated in a substance so it has a - charge.
• the gel is covered in a buffer solution containing ions - conducts electrical current
• samples are loaded at the end of the gel and power source is applied
• negatively charged proteins move towards positive end
• smaller proteins move faster
• a protein ladder is put on the gel as well as a standard to compare sample to allow protein to be identified

Question 12

separating proteins in a sample (charge) - electrophoresis

Answer 12

• sample of amino acids is placed in the middle of the gel
• gel is in a bath with a particular pH value
• power source is used to supply a positive and negative electrode at ends of the gel
• depending on the isoelectric points - they will be - or +
• positively charged amino acids will move towards the negative electrode
• if more than one amino acid is + or - they can be assessed on size