Natural Products Training Laboratory

The University of Mississippi

5-Day Course Outline

Day 1: Botanical Authentication

  1. Taxonomy & Identification
    1. Plant taxonomy
    2. Botanical nomenclature
    3. Plant morphology
    4. Plant anatomy
    5. Herbarium/Voucher specimens
    6. Species identification
  2. Adulteration
    1. Adulterants and substitutes
  3. Authentication of Botanicals
    1. Organoleptic
    2. Macroscopy
    3. Microscopy
  4. GC-MS (technique and applications for botanicals/dietary supplements)
  5. HPTLC Technique and Lab Session – Overview of technology and entire process
    1. Preparation of samples and developing solvent (mobile phase)
    2. Plate marking
    3. Spotting a plate
    4. Placing plate in development chamber
    5. Conditioning development chamber
    6. Development of plate
    7. Visualization and interpretation
    8. Calculations of RF values
    9. Plate derivitization and detection

 

Day 2: Sample Preparation and Analysis

  1. HPTLC Vs. HPLC
    1. Comparison
    2. Advantages and disadvantages
  2. Sample preparation
    1. Types of samples
    2. Sample handling
    3. Sample preparation techniques
  3. Modern separation techniques (HPLC, UHPLC)
    1. Theory
    2. Comparison
    3. Practical applications
  4. Detection techniques (UV-VIS, ELSD, MS)
    1. Comparison
    2. Key applications for natural products
  5. Columns/Column chemistry
    1. Column dimensions
    2. Particle/Pore size
    3. Stationary phases
  6. Types of analysis
    1. Qualitative (Fingerprinting)
    2. Quantitative (Marker compounds)
  7. Method transfer
    1. Overview
  1. HPLC Lab Session

 

Day 3: Data Processing

  1. Data processing
    1. Retention time
    2. Peak shape
    3. Resolution
    4. Spectral data
  2. Peak homogeneity
    1. Mass analysis window
  1. Automated Custom Calculations
    1. Milligram per capsule/tablet (For Liquids, mg/ml)
    2. Weight per serving
    3. Weight per daily dose
  1. Acceptance Limits
    1. Within specification
  1. UV Spectral Matching (Libraries)
    1. Overview
    2. Theory
    3. Benefits
    4. Practical applications
  2. Mass Detection
    1. Overview
    2. Benefits
    3. Practical applications

 

Day 4: MS Theory

  1. Introduction
  2. Theory & Overview
    1. Ionization
    2. Ionization modes
    3. Quadrupoles
    4. Collision cells
    5. Fragmentation
    6. Adducts
    7. Single-quad Vs. Tandem-quad
    8. Full-scan vs. SIR vs. MRM
    9. Matrix effects (ion suppression or enhancement)
  3. Empowering Technology
    1. RADAR – evaluation of matrix background/interference
    2. PIC/S – product/ion scans for MRM confirmation
  4. Acquisition Modes
    1. LC/MS
    2. Direct infusion
    3. ASAP probe
    4. TLC/MS
    5. DART
  5. Method Development
    1. Optimization (automated using Intellistart)
  1. Practical Applications
    1. Mass confirmation
    2. Ms Quant for non-chromophore compounds
    3. Ms Quant for trace-level compounds
    4. Ms Quant in highly complex matrices

 

Day 5: MS Quantification

  1. Calibration
    1. Internal standard
    2. Deuterated standards
    3. External standards
    4. Standard addition
  2. Quantification
    1. Linear range
    2. Quantifier/Qualifier ions
    3. Ion ratio
    4. Limits of quantification
    5. Limits of detection
  3. Key Applications
    1. Pesticide residue analysis (Targeted)
    2. Adulteration screening (PDE-5 Inhibitors, Yohimbe, etc.)
    3. Quantification of minor compounds

Closing Remarks: Dr. Khan