- Question 1: What is Full form of HPLC? …
- Question 2: What is HPLC Principle? or What is Principle of Chromatography? …
- Question 3: What is General chapter number of Chromatography? …
- Question 4: How many types of chromatography?
HPLC interview Question and Answer | Pharmabeej
The interview questions cover questions from basic to advanced levels of technical aspects. These HPLC interview questions and answers will help crack an interview, enhance your knowledge, and also be helpful for the interviewer who is involved in the recruitment process.
In normal phase chromatography it is recommended to avoid using very dry solvents. Most preferable approach to solve the equilibration problem of silica with water is to use solvents that are “half-saturated” with water. It can be prepared by saturating a provided volume of hydrophobic solvent with water and mixing it 1:1 with “dry” solvent. This will facilitate quick equilibration of the column.
You will find interview questions and answers on components of an HPLC system, isocratic and a gradient HPLC system, types of gradient mixing, function of the HPLC column, types of sample injector, function of the HPLC column oven, commonly used detector in HPLC systems, types of detectors used in HPLC systems, typical HPLC startup procedure, procedure to flush the new reversed-phase column, checkpoints before starting an HPLC analysis, relative retention time (RRT) in liquid chromatography, ‘fronting’ or ‘leading’ in liquid chromatography, peak symmetry, signal-to-noise ratio, procedures for mobile phase degassing, advantages of acetonitrile in the mobile phase, advantages of methanol in the mobile phase, reasons for variable Retention Times (RT) in HPLC analysis, and many other relevant and valuable HPLC interview questions and answers.
You will find it much more enjoyable while going through these interview questions and answers. So enjoy learning and best of luck with your interview! Happy Learning.
HPLC interview questions are the most commonly asked topic during the quality control laboratory interview for the pharmaceutical industry. HPLC is the pharmaceutical industry’s most widely used technique of analysis; therefore, most of the interview questions and answers are around HPLC technique and HPLC troubleshooting.
Question 19: Which calibration standard used for HPLC Calibration?Answer: Caffein standard is used for HPLC calibration.
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.
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 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.
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.
Method validation in chromatography must be carried out as per the validation protocol is given by the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) or The United States Pharmacopeia (USP) and British Pharmacopoeia (BP). The protocol includes the process and criteria of acceptance for all protocol.
Column chromatography is a general technique that is used to separate the compounds from the complex mixtures. It can use small or large-scale column chromatography to separate and purify the analytes. Column chromatography has two phases, a mobile phase (liquid) and a stationary phase (solid). The sample mixture of compounds travels with the mobile phase through the stationary phase and basis of different degrees of adhesion it separates.The column chromatography works on the principle of adsorption. There are four different column chromatography types for several applications and that work on different mechanisms.
Its a method validation parameter. The reference standard is a preferred technique by direct comparison accuracy. In this technique of validation parameter, perform recovery study by selecting three different concentrations like 0 % (0 % is your standard) 80 %, 100 % and 120 %. Compare to a standard sample and recovery should be in the range of 99 to 101 %.
Specificity is a method validation parameter, Spike your sample in addition with degradation products, solvents, chemicals and solvents used to perform analysis, hence all analytes should be separate and minimum resolution have 2.0. and another one is to spike samples of marketed preparation, it should be separate having RSD ≤ 2 %.
In this parameter of method validation, the range of 0-150% of the expected level of analysis should be covered. The method should display linearity in the desired range. For the linearity purpose, prepare six samples in desire range to take linearity and correlation coefficient value should be minimum R² = 0.99.
Answer: Dead volume is the volume of the HPLC system between points of injection to point of detection.
Answer: Related substance is used to determine known as well as unknown impurity while chromatography purity is used to determine only known impurities from the sample.
Answer: Dwell volume is a volume of gradient HPLC system between mixing chamber in column inlet.
Answer: – C8 has 8 carbon atoms and C18 has 18 carbon atoms, C18 is more hydrophobic than C8 due to that C18 has a long retention time and C8 has a sort retention time (early).
Answer: HPLC is a separation technique of components from the mixer by using solid stationary phase and liquid mobile phase.
FAQ
What are the different types of columns used in HPLC?
The specific intermolecular interactions between the molecules of a sample and the packing material define their time “on-column”. Hence, different constituents of a sample are eluted at different times. Thereby, the separation of the sample ingredients is achieved.
Why does RSD fail in HPLC?
- Normal Phase Columns.
- Reverse Phase Columns.
- Ion Exchange Columns.
- Size Exclusion Columns.
