HPLC Interview Questions

HPLC Interview Questions

Question 1: What is Full form of HPLC?

Answer: HPLC stands for High-performance Liquid Chromatography or High-pressure Liquid Chromatography.

Question 2: What is HPLC Principle? or What is Principle of Chromatography?

Answer: It is a chromatographic technique used to separate the components from a mixture of a solution by using solid stationary phase and liquid mobile phase.

Question 3: What is General chapter number of Chromatography?

Answer: USP General chapter Number <621>

Question 4: How many types of chromatography?

Answer: There are 12 types of chromatography.

Question 5: What are the names of different types of chromatography?

Answer:

3. Thin Layer Chromatography (TLC)
4. Paper Chromatography
5. Column Chromatography
6. Affinity Chromatography
7. Fast Protein Liquid Chromatography
8. Supercritical Fluid Chromatography
9. Reverse-phase Chromatography
10. Countercurrent Chromatography
11. Two Dimensional Chromatography
12. Pyrolysis Gas Chromatography

Question 6: What are the types of Column Chromatography?

Answer: There are 5 types of chromatography method where column is used.
1. Liquid Chromatography
2. Gas Chromatography
3. Ion Exchange Chromatography
4. Size Exclusion Chromatography
5. Chiral Chromatography

Question 7: What is Column Chromatography Principle?

Answer: The column chromatography principle is, Column chromatography is fundamentally based on the adsorption of solutes from a solution by means of a stationary phase, which then separates each component of the mixture.

Question 8: What is Reverse phase chromatography?

Answer: Mobile phase is more polar than the stationary phase is called reserve phase chromatography.

Question 10: What is Chiral Chromatography?
Answer: It is a chromatography to determine the content of chiral isomers whether it is in normal or reverse phase.

Question 11: What is Column?

Answer: Column is a still tube which contains a stationary phase.

Question 12: What are the component of HPLC?

Answer: There are 6 types of components,
1. Reservoir
2. Pump
3. Sample Components
4. Column Components
5. Detector
6. Recorder

Question 13: What are the types of Detectors used in Chromatography?

Answer: There are 6 detectors are used in HPLC based on their usage,
1. UV detector
2. PDA Detector
3. Fluorescence detector
4. Conductivity detector
5. Refractive Index Detector
6. Light Scattering Detector

Question 14: What is Gradient run?

Question 15: What is Isocratic run?

Answer: Mobile phase remains the same throughout the run. As mentioned above 20 min run time.The mixed (Composition of buffer + aqueous solution) mobile phase is used in the isocratic run.

Question 16: What is Linear gradient?

Answer: Mobile phase remains unchanged throughout the run by using two different reservoirs.
Understand with the below linear-gradient table. Mobile phase A and Mobile phase B is in the same proportion up to 20 mins.

Question 17: What is Retention time (RT)?

Answer: Retention time is a time between injection and the appearance of the peak maxima.

Question 18: What is Relative retention time (RRT)?

Answer: RRT is a measure of the difference of affinities of two compounds for the stationary phase.

Question 19: Which calibration standard used for HPLC Calibration?
Answer: Caffein standard is used for HPLC calibration.

Question 20: Why Caffein standard is used for HPLC calibration?

3. Readily available in market
4. It shows two maxima and one minima at 205 nm,273 nm, and 245 nm
respectively.

Question 21: What type of material is most commonly used in the stationary phase?

Answer: Silica gel is most commonly used material in stationary phase.

Question 22: Why Silica gel is used in stationary phase?

Answer: Silica gel is inert material and does not react with mobile phase.

Question 23: What is Flow diagram of the HPLC System?

Answer:
1. Solvent reservoir
2. Pump
3. Degasser
4. Mixing valve
5. Guard column
6. Sample injector
7. Column
8. Detector
9. Recorder
10. Outlet