5-Day Course Outline
Day 1: Botanical Authentication
- Taxonomy & Identification
- Plant taxonomy
- Botanical nomenclature
- Plant morphology
- Plant anatomy
- Herbarium/Voucher specimens
- Species identification
- Adulteration
- Adulterants and substitutes
- Authentication of Botanicals
- Organoleptic
- Macroscopy
- Microscopy
- GC-MS (technique and applications for botanicals/dietary supplements)
- HPTLC Technique and Lab Session – Overview of technology and entire process
- Preparation of samples and developing solvent (mobile phase)
- Plate marking
- Spotting a plate
- Placing plate in development chamber
- Conditioning development chamber
- Development of plate
- Visualization and interpretation
- Calculations of RF values
- Plate derivitization and detection
Day 2: Sample Preparation and Analysis
- HPTLC Vs. HPLC
- Comparison
- Advantages and disadvantages
- Sample preparation
- Types of samples
- Sample handling
- Sample preparation techniques
- Modern separation techniques (HPLC, UHPLC)
- Theory
- Comparison
- Practical applications
- Detection techniques (UV-VIS, ELSD, MS)
- Comparison
- Key applications for natural products
- Columns/Column chemistry
- Column dimensions
- Particle/Pore size
- Stationary phases
- Types of analysis
- Qualitative (Fingerprinting)
- Quantitative (Marker compounds)
- Method transfer
- Overview
- HPLC Lab Session
Day 3: Data Processing
- Data processing
- Retention time
- Peak shape
- Resolution
- Spectral data
- Peak homogeneity
- Mass analysis window
- Automated Custom Calculations
- Milligram per capsule/tablet (For Liquids, mg/ml)
- Weight per serving
- Weight per daily dose
- Acceptance Limits
- Within specification
- UV Spectral Matching (Libraries)
- Overview
- Theory
- Benefits
- Practical applications
- Mass Detection
- Overview
- Benefits
- Practical applications
Day 4: MS Theory
- Introduction
- Theory & Overview
- Ionization
- Ionization modes
- Quadrupoles
- Collision cells
- Fragmentation
- Adducts
- Single-quad Vs. Tandem-quad
- Full-scan vs. SIR vs. MRM
- Matrix effects (ion suppression or enhancement)
- Empowering Technology
- RADAR – evaluation of matrix background/interference
- PIC/S – product/ion scans for MRM confirmation
- Acquisition Modes
- LC/MS
- Direct infusion
- ASAP probe
- TLC/MS
- DART
- Method Development
- Optimization (automated using Intellistart)
- Practical Applications
- Mass confirmation
- Ms Quant for non-chromophore compounds
- Ms Quant for trace-level compounds
- Ms Quant in highly complex matrices
Day 5: MS Quantification
- Calibration
- Internal standard
- Deuterated standards
- External standards
- Standard addition
- Quantification
- Linear range
- Quantifier/Qualifier ions
- Ion ratio
- Limits of quantification
- Limits of detection
- Key Applications
- Pesticide residue analysis (Targeted)
- Adulteration screening (PDE-5 Inhibitors, Yohimbe, etc.)
- Quantification of minor compounds
Closing Remarks: Dr. Khan