Lecture 03, 4

• What is chromatography used for?

  Separation of mixtures into components
• What are the two phases in chromatography?

  Mobile phase and stationary phase
• What is retention time in chromatography?

  Time taken for a compound to pass through
• What does column efficiency refer to in chromatography?

  How well a column separates compounds

  
• What is plate theory in chromatography?

  A model to describe column efficiency
• What is selectivity in chromatography?

  Ability to separate different compounds
• What is resolution in chromatography?

  Degree of separation between peaks
• What are the types of chromatography mentioned?

  TLC, HPLC, GC
• What are the types of chromatography separation?

  • Adsorption
  • Partition
  • Ion exchange
  • Size exclusion
• What is the difference between normal and reverse phase chromatography?

  Normal phase uses polar stationary, reverse uses non-polar
• What is adsorption chromatography?

  Separation based on sample components adsorbing to solid
• What is partition chromatography?

  Separation based on liquid stationary phase on solid
• What is ion exchange chromatography?

  Separation based on ion exchange equilibria
• What is size exclusion chromatography?

  Separation based on size of solvated molecules

  
• How does HPLC work?

  Separates compounds using high pressure liquid flow

  
• What is column selectivity in HPLC?

  Ability of the column to separate analytes
• Why is chromatography important in medicine development?

  Ensures quality of pharmaceutical products
• What types of compounds can HPLC separate?

  Volatile, non-volatile, organic, inorganic
• What is the role of the solvent in HPLC?

  Acts as the mobile phase for separation
• What is isocratic elution in HPLC?

  Constant mobile phase composition throughout run
• What is gradient elution in HPLC?

  Changing mobile phase composition over time
• What is the purpose of the injector in HPLC?

  To introduce sample into the mobile phase flow
• How are chromatographic techniques classified?

  • Based on interaction between mobile and stationary phase
  • Types: Adsorption, Partition, Ion exchange, Size exclusion
• What is adsorption chromatography?

  Solid stationary phase adsorbs sample components

  
• What is partition chromatography?

  Liquid stationary phase on inert solid
• What is the mechanism of ion exchange chromatography?

  Based on ion exchange equilibria
• How does size exclusion chromatography work?

  Separates molecules by size using a sieve-like structure

  
• What is normal phase chromatography?

  Uses polar stationary phase and non-polar mobile phase
• What is reverse phase chromatography?

  Uses non-polar stationary phase and polar mobile phase

  
• How do polar analytes behave in reverse phase chromatography?

  They elute quickly and are not retained
• What is the role of pumps in HPLC?

  Deliver mobile phase at constant flow rate
• What is the function of detectors in HPLC?

  Convert analyte response into electrical signal
• What is the importance of solvent choice in HPLC?

  Samples must be soluble in the mobile phase
• What are the characteristics of HPLC columns?

  Made from stainless steel, bonded phases
• How does particle size affect HPLC?

  Smaller particles improve separation efficiency
• What is the calibration curve method in HPLC?

  Method to determine plate number for identification
• What is the standard addition method in HPLC?

  Adding known standard to sample for analysis
• What are the advantages of HPLC?

  Speed, high resolution, sensitivity, accuracy
• What are examples of detectors used in HPLC?

  UV detectors, MS detectors, fluorescence detectors
• What is gas chromatography (GC)?

  Technique for separating volatile compounds