Chromatography

Question 1

What are the three applications of chromatography

Answer 1

Quantitation

Identification

Purification

Question 2

What is extraction chromatography

Answer 2

You have 2 immiscible phases, the solutes move to each diff phase depending on their solubility’s

The strength of their interaction with each other determines where they go

(Ex. If lower layer is water, more polar things go in the lower layer)

Question 3

What are the 3 ways the extraction can be adjusted to move things into different phases

Answer 3

Ph

Chelators

Surfactants

Question 4

How does ph affect extractions

Answer 4

Affects the efficiency,

if protonated or deprotonated the solvent goes to less or more polar layer because the charge has changed

Question 5

How do chelators affect extractions

Answer 5

They change the solubility of metal ions

If the ions are chelated, they can go into the organic (nonpolar) layer by decreasing the charge/polarity of the ion

Makes things neutral/less polar

Question 6

How do surfactants affect extractions

Answer 6

They interact with the hydrophobic and hydrophilic phases by making micelles and liposomes

These surfactants can force things to go into the nonpolar layer by surrounding the particle and making it hydrophobic (less polar)

Question 7

What is regular chromatography

Answer 7

Similar to extractions

There is a stationary phase and a mobile (eluent) phase that the solvent can interact with differently depending on the strath of their interactions with thhat phadr

If the eluent is flowing, things that interact with it will flow too

Question 8

Do things have zero interaction with either mobile or stationary phase

Answer 8

No there’s always some interaction but just less

Question 9

If something interacts less with stationary phase when does it get seen in a column

Answer 9

First solvent (mobile phase) always elutes first then the thing that interacts least with the stationary phase comes after

Question 10

If something elites out after adding a diffent solvent than before when it’s wasn’t eluding what does this mean

Answer 10

The new solvent and the thing have stronger interactions with each other then the old solvent

Question 11

What does the types of chromatography depend on and what are the 5 types of chromatography

Answer 11

Depends on the type of stationary phase used

Adsorption chromatography

Partition

Ion exchange

Size exclusion

Affinity

Question 12

What is adsorption chromatography

Answer 12

The solutes interact with the surface of a solid stationary phase

Question 13

What is partition chromatography

Answer 13

The solutes dissolve into a liquid stationary phase

Question 14

What is ion exchange chromatography

Answer 14

The solute has a chemical reaction with a resin stationary phase instead of just interacting

It exchanges ions

Question 15

What is size exclusion (gel) chromatography

Answer 15

The solutes get filtered through a mesh gel or interact with the pores of the gel

Larger travel slower smaller travel faster

Question 16

What is affinity chromatography

Answer 16

The molecules of the solute has specific antibody or antigens or DNA/RNA pairings that bind to the stationary phase via chemical reactions

Question 17

What are chromatograms

Answer 17

A plot of signal vs time from the detector as the solutes flow down the elution column

What ever elites first shows up on the graph first

The area under the peaks is proportional to their respective amount of solute

Question 18

What is retention time

Answer 18

The amount of time it took for the sample to elute

Question 19

TM TR TR PRIME IN TABLET pics

Answer 19

WRITE ON FOLUMA SHEET

Question 20

What is separation factor

Answer 20

The ratio of how far apart the peaks are in a chromatogram (the adjusted retention time of 2 over the adjusted retention time of 1)

It’s independent of flow rate which means it’s good for comparing across experiments

It’s symbol is alpha

Question 21

What is retention factor (K)

Answer 21

How fast the peak goes vs the fastest (unretained) compounds

The adjusted retention time over the TM

Bigger K = slower

Question 22

Measuring peak resolution

Answer 22

Slide 16 in tablet pics

Question 23

What resolution is best to see peaks and quantify them in a chromatogram

Answer 23

1.50

And the peaks a 6 sigmas apart

Question 24

What happens to the peaks when there’s a larger retention time

Answer 24

The resolution of the peaks gets worse because the bands widen

They widen because they diffuse inside the columns along with having concentration gradients

Question 25

Slide 18 onwards

Answer 25

Do the cards