Chromatography Flashcards
Father of chromatography
Tswett
What is the polarity of stationary phase and mobile phase
Stationary phase is polar and mobile phase nonpolar
What kind of paper chromatography is
Paper chromatography is partition chromatography depends on absorption and is liquid liquid chromatography
Thin layer chromatography is which type of chromatography
Solid liquid chromatography and adsorption chromatography
What is the matrix of TLC? Type of bond?
Silica gel or alumina on glass metal or plastic. Non covalent bond
Why is descending chromatography is preferred?
Works for little Rf value too
Adsorptive effect of polar compounds due to?
Hydroxyl group
Applications of paper chromatography
Determining purity
Analysis of mixture
Detecting drugs
Stationary and mobile phase of column chromatography
Stationary phase: Alumina, charcoal, silica, diatomaceous earth
Mobile phase: Chloroform, ethyl ether, and hexane
Name the hydrated polymer stationery phase of size exclusion chromatography is made of
Sephadex, sepharose and biogel
What is exclusion limit
When the smallest particle cannot enter the beads is called exclusion limit
Formula of total volume of size exclusion chromatography
Vt = V0 + Vi + Vg
Give formula of elution volume
Ve = Vo + KdVi
What is the difference between gel permeation and gel filtration
Gel permeation uses organic solvent and gel filtration uses aqueous solvent
Hydroxyapetite chromatography uses what
Calcium phosphate beads
Why is hydroxyapetite chromatography used
To separate proteins and nucleic acid
Why is ion exchange chromatography used
To separate ion and polar molecules
What is the bond in ion exchange chromatography
Reversible electrostatic interaction
What is cation exchanger beads and give example
The beads that attract positively charge molecules and examples are carboxyl methyl cellulose and sulfopropyl
What is anion exchanger beads and give example
Those who attack negatively charge molecules. The examples are diethyl amino ethyl cellulose.
What is the elusion order of ion exchanger chromatography
For cation exchange the lowest pI will elute first and for anion exchange the highest pI will elute first
Why is affinity chromatography used
To purify biomolecule
What kind of interaction does ligand and target molecule have
Reversible interaction
Name the target molecule of the following
1. Substrate
2. Avidine/ Strepavidin
3. Ni Metal
4. Lectin
5. Antibody
- Enzyme
- Biotine
- Poly histadine
- Carbohydrates
- Antigen
What are the ligands use to bind DNA sequence
ds oligonucleotide
What type of interaction is found in reverse phase chromatography
Hydrophobic interaction
What is gradient elusion also called
Solvent programming
What happens in solvent programming. Applications
The composition of mobile phase keeps changing. Helps in determining solvent purity mixture of polar and nonpolar liquid
What is the composition of stationary and mobile phase in hydrophobic interaction chromatography
Stationary phase is made of agarose with propyl or octyl group and mobile phase is polar and the polarity decreases
What is the pressure used in high performance liquid chromatography
1000 to 6000 psi
How is the resolution of high performance liquid chromatography high
Because of increased colum size and small packing material
What are the bulk property and general purpose detector in HPLC
Bulk property is refractive index detector. General purpose detector is evaporative light scattering detector
What is the solute property detector
UV Vis detector, photo diode array detector and fluorescence detector
Why is gas chromatography used
To analyse volatile substance
Which is the chromatography where the mobile phase doesn’t interact with the analyte
Gas chromatography
Examples of carrier gas
Helium, nitrogen, argon and hydrogen
Selectivity factor
The ratio of Rf value that tells us how well the two samples are distinguished. Higher the value greater the resolution
Van De Emter Plot
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
Theoretical plate
The more is the height of the plate the better the resolution. The solute in these plates are in equilibrium
Retention time
The time of injecting analyte till it reaches the detector
Retention index. What is it mainly used for?
Comparison of different retention time. Gas chromatography
Column bleeding
Thermal breakdown or elusion of stationery phase
Which chromatography technique separate neomycin sulfate
TLC
Chromatography Technique that helps in protein protein interaction
Pull down affinity chromatography
Which chromatography technique helps in molecular mass determination
Size exclusion chromatography
