Course Schedule date:
| Monday | |Tuesday | | Wednesday | | Thursday | Friday | (8.0 PM to 10 PM; IST)
Module - 1
- What is the Analytical method development?
- Why are chromatographic techniques needed?
- A Birds Eye View on Commonly used chromatographic techniques
- HPLC and its components
- How to read a chromatogram?
- RT, RRT, area response and peak height
- Types of chromatography
- HPLC operating parameters
- Basic separation mechanism
- Detectors
Module - 2
- Chromatographic terms and System suitability test parameters (K, N, R, T, As, HETP, RSD, DL, QL. As)
- Mobile Phase Chemistry
- Type of Elution mode
- Column chemistry
- A bird’s Eye view on different Analytical tests
- Advantages and disadvantages of HPLC
- Role NPC, RPC and IP-RPC in HPLC method development
- pH and its role in HPLC method development
- pKa and its role in HPLC method development
- Role of polarity in HPLC method development
Module - 3
- How to equilibrate the column faster?
- Elution mode selection: Isocratic vs Gradient: advantages and disadvantages
- Modes of calculation
- How to prepare solutions of different concentrations (mcg, ppm etc)
- Different components of the HPLC method
- Precautions required during HPLC analysis
- Compounds classification based on polarity
- HPLC Separation principle
- A Bird’s Eye view on HPLC separation mechanism
- How to predict elution order?
Module - 4
- RFT approach in HPLC method development
- Method development steps
- Sample preparation procedures
- Chromatographic mode selection
- Column selection procedure
- Mobile phase selection procedure
- Selection of buffer (type and concentration)
- Selection of modifier
- Solvent selection and optimization
- Detector Selection and role of PDA detector in HPLC method development
- Calculation mode selection
- How to optimise retention time?
- Method Optimization in gradient and isocratic mode(Temperature, sample concentration, injection volume, flow rate, wavelength and separation)
Module - 5
- Characteristics of a newly developed HPLC method
- HPLC Method verification
- Common HPLC troubleshooting
- How to reduce noise?
- How to deal with over pressure in the column?
- How to deal with column equilibration issue?
- How to deal with SST failure issue?
- How to deal with mobile phase precipitation?
- How to increase the HPLC column life?
- Brief discussion on column care
- Separation of a mixture of nonpolar, polar acidic, polar basic, strongly acidic and strongly basic compounds
- How to develop a cost-effective method
- How to develop HPLC-MS compatible method?
- How to peak integration?
- How to check peak purity?
- Case Studies
- Exercise
- Doubt Clearances
Bonus
- How to deal with fronting and tailing in the peak?
- How to deal with variation in retention time?
- How to deal with variation in area response?
- How to deal with Ghost Peak?