Chromatography

Question 1

Father of chromatography

Answer 1

Tswett

Question 2

What is the polarity of stationary phase and mobile phase

Answer 2

Stationary phase is polar and mobile phase nonpolar

Question 3

What kind of paper chromatography is

Answer 3

Paper chromatography is partition chromatography depends on absorption and is liquid liquid chromatography

Question 4

Thin layer chromatography is which type of chromatography

Answer 4

Solid liquid chromatography and adsorption chromatography

Question 5

What is the matrix of TLC? Type of bond?

Answer 5

Silica gel or alumina on glass metal or plastic. Non covalent bond

Question 6

Why is descending chromatography is preferred?

Answer 6

Works for little Rf value too

Question 7

Adsorptive effect of polar compounds due to?

Answer 7

Hydroxyl group

Question 8

Applications of paper chromatography

Answer 8

Determining purity
Analysis of mixture
Detecting drugs

Question 9

Stationary and mobile phase of column chromatography

Answer 9

Stationary phase: Alumina, charcoal, silica, diatomaceous earth
Mobile phase: Chloroform, ethyl ether, and hexane

Question 10

Name the hydrated polymer stationery phase of size exclusion chromatography is made of

Answer 10

Sephadex, sepharose and biogel

Question 11

What is exclusion limit

Answer 11

When the smallest particle cannot enter the beads is called exclusion limit

Question 12

Formula of total volume of size exclusion chromatography

Answer 12

Vt = V0 + Vi + Vg

Question 13

Give formula of elution volume

Answer 13

Ve = Vo + KdVi

Question 14

What is the difference between gel permeation and gel filtration

Answer 14

Gel permeation uses organic solvent and gel filtration uses aqueous solvent

Question 15

Hydroxyapetite chromatography uses what

Answer 15

Calcium phosphate beads

Question 16

Why is hydroxyapetite chromatography used

Answer 16

To separate proteins and nucleic acid

Question 17

Why is ion exchange chromatography used

Answer 17

To separate ion and polar molecules

Question 18

What is the bond in ion exchange chromatography

Answer 18

Reversible electrostatic interaction

Question 19

What is cation exchanger beads and give example

Answer 19

The beads that attract positively charge molecules and examples are carboxyl methyl cellulose and sulfopropyl

Question 20

What is anion exchanger beads and give example

Answer 20

Those who attack negatively charge molecules. The examples are diethyl amino ethyl cellulose.

Question 21

What is the elusion order of ion exchanger chromatography

Answer 21

For cation exchange the lowest pI will elute first and for anion exchange the highest pI will elute first

Question 22

Why is affinity chromatography used

Answer 22

To purify biomolecule

Question 23

What kind of interaction does ligand and target molecule have

Answer 23

Reversible interaction

Question 24

Name the target molecule of the following
1. Substrate
2. Avidine/ Strepavidin
3. Ni Metal
4. Lectin
5. Antibody

Answer 24

1. Enzyme
2. Biotine
3. Poly histadine
4. Carbohydrates
5. Antigen

Question 25

What are the ligands use to bind DNA sequence

Answer 25

ds oligonucleotide

Question 26

What type of interaction is found in reverse phase chromatography

Answer 26

Hydrophobic interaction

Question 27

What is gradient elusion also called

Answer 27

Solvent programming

Question 28

What happens in solvent programming. Applications

Answer 28

The composition of mobile phase keeps changing. Helps in determining solvent purity mixture of polar and nonpolar liquid

Question 29

What is the composition of stationary and mobile phase in hydrophobic interaction chromatography

Answer 29

Stationary phase is made of agarose with propyl or octyl group and mobile phase is polar and the polarity decreases

Question 30

What is the pressure used in high performance liquid chromatography

Answer 30

1000 to 6000 psi

Question 31

How is the resolution of high performance liquid chromatography high

Answer 31

Because of increased colum size and small packing material

Question 32

What are the bulk property and general purpose detector in HPLC

Answer 32

Bulk property is refractive index detector. General purpose detector is evaporative light scattering detector

Question 33

What is the solute property detector

Answer 33

UV Vis detector, photo diode array detector and fluorescence detector

Question 34

Why is gas chromatography used

Answer 34

To analyse volatile substance

Question 35

Which is the chromatography where the mobile phase doesn't interact with the analyte

Answer 35

Gas chromatography

Question 36

Examples of carrier gas

Answer 36

Helium, nitrogen, argon and hydrogen

Question 37

Selectivity factor

Answer 37

The ratio of Rf value that tells us how well the two samples are distinguished. Higher the value greater the resolution

Question 38

Van De Emter Plot

Answer 38

Explain the relationship between the flow rate and the efficiency of separation of the analyte. The lower the value of h, the better the separation

Question 39

Theoretical plate

Answer 39

The more is the height of the plate the better the resolution. The solute in these plates are in equilibrium

Question 40

Retention time

Answer 40

The time of injecting analyte till it reaches the detector

Question 41

Retention index. What is it mainly used for?

Answer 41

Comparison of different retention time. Gas chromatography

Question 42

Column bleeding

Answer 42

Thermal breakdown or elusion of stationery phase

Question 43

Which chromatography technique separate neomycin sulfate

Answer 43

TLC

Question 44

Chromatography Technique that helps in protein protein interaction

Answer 44

Pull down affinity chromatography

Question 45

Which chromatography technique helps in molecular mass determination

Answer 45

Size exclusion chromatography