20+HPLC Interview Questions and answers in pharma

HPLC Interview Questions And Answers

If the interview is conducted for quality control in the pharma or research and development department, then HPLC interview questions and answers are a must. HPLC is an instrument that separates the different components from the solution mixture by using two different phases i.e. stationary phase and mobile phase.

Some basic HPLC interview questions and answers are collected. Here we will understand all those interview questions for HPLC.

Question 1: What is the Full form of HPLC?

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

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

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

Question 3: What is the 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?


  1. Gas Chromatography (GC)
  2. High-Pressure Liquid Chromatography (HPLC)
  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 methods where the column is used.

  1. Liquid Chromatography
  2. Gas Chromatography
  3. Ion Exchange Chromatography
  4. Size Exclusion Chromatography
  5. Chiral Chromatography

Question 7: What is the 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 9: What is Normal phase chromatography?

Answer: Stationary phase is more polar than the mobile phase called normal 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 a Column?

Answer: A column is a still tube that contains a stationary phase.

Question 12: What is the Stationary Phase?

A: The packing material for the column, which is the immobile phase in the chromatographic process, is known as the stationary phase.

Question 13: What is the Mobile Phase?

A: The liquid that transports the solute through the column is known as the mobile phase.

Question 14: What are the components of HPLC?

Answer: There are 6 types of components,

  1. Reservoir
  2. Pump
  3. Sample Components
  4. Column Components
  5. Detector
  6. Recorder

Question 15: 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 16: What is Gradient run?

Answer: Mobile phase composition varies over run time (Gradient Programm). understand with 20 min run time.

Run Time in minMobile phase AMobile Phase B
Gradient Run

Also Read:

ICH Q10 in pharmaceutical quality system

Difference Between 21 CFR part 11 and EU Annex 11

Question 17: 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 18: 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.

Run Time in minMobile phase AMobile Phase B
Linear Gradient

Question 19: What is Retention time (RT)?

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

Question 20: What is Relative retention time (RRT)?

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

Question 21: Which calibration standard is used for HPLC Calibration?

Answer: The caffeine standard is used for HPLC calibration.

Question 22: Why caffeine standard is used for HPLC calibration?


  1. It is Durable
  2. It is very stable.
  3. Readily available in market
  4. It shows two maxima and one minima at 205 nm,273 nm, and 245 nm respectively.

Question 23: What type of material is most commonly used in the stationary phase?

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

Question 24: Why Silica gel is used in the stationary phase?

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

Question 25: What does inert gas mean?

A: An inert gas is a gas that has no reaction with anything (Non-reactive). Generally, inert gases don’t react with anything or anyone, which is why they’re called inert gases.

Question 26: What is the Flow diagram of the HPLC System?


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

Q27: What does mean by Frit in HPLC?

A: Frit is a device that provides a coarse filtration of the HPLC eluent and keeps small particles from entering the high-performance liquid chromatography system. It is commonly inserted before pre-columns.

HPLC Interview Questions And Answers

These are the most common HPLC Interview Questions And Answers ask by the interviewer while interviewing.

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