Unit 4: Phytochemistry and Analytical Pharmacognosy

Active principles and methods of their testing

Active Principles (Phytochemicals/Secondary Metabolites)

Active Principles: These are the specific chemical compounds found in a medicinal plant that are responsible for its therapeutic (healing) effect.

They are also called secondary metabolites because they are not essential for the plant's basic survival (like primary metabolites - carbs, proteins), but are instead produced for defense, attraction, or signaling.

Major classes of active principles include:

Methods of their Testing (Extraction & Analysis)

Testing involves extracting these compounds from the plant and then identifying/quantifying them.

  1. Extraction: The plant material (crude drug) is dried, powdered, and then mixed with a solvent (e.g., alcohol, water, ether) to dissolve the active principles. The liquid extract is then filtered and concentrated.
  2. Analysis & Testing:
    • Phytochemical Screening: Simple, qualitative test-tube reactions to check *if* a class of compound is present (see topic 6).
    • Chromatography: The primary tool for separation and identification.
      • TLC (Thin Layer Chromatography): A simple, fast method to separate compounds on a coated plate. Used for identification.
      • HPLC (High-Performance Liquid Chromatography): A sophisticated, high-pressure, quantitative method. It can separate, identify, and tell you *how much* of a compound is present. This is the industry standard for quality control.
    • Spectroscopy: Used to determine the chemical structure of an isolated compound (e.g., UV-Vis Spectroscopy, Mass Spectrometry).

Management of herbal medicines

This refers to the proper handling of medicinal plants from collection to consumption to ensure safety, quality, and efficacy.

Drug adulteration (types)

Adulteration: The practice of substituting the original crude drug with other, similar-looking substances that are of inferior quality, less effective, or harmful. This is usually done for financial gain.

Types of Adulteration

Methods of drug evaluation

Drug evaluation is the process of quality control to confirm the identity, purity, and quality of a crude drug and to detect adulteration.

  1. Organoleptic Evaluation:
    • Uses the sensory organs (sight, smell, taste, touch).
    • Checks the drug's color, odor, taste, size, shape, and texture.
    • Example: Smelling cinnamon bark, tasting a licorice root for sweetness.
  2. Microscopic Evaluation:
    • Uses a microscope to examine the fine cellular structure of the drug.
    • Looks for specific, diagnostic features (e.g., types of stomata, trichomes (hairs), stone cells, calcium oxalate crystals).
    • Example: Identifying senna leaf powder by its unique unicellular trichomes.
  3. Physical Evaluation:
    • Uses physical constants.
    • Examples: Measuring moisture content (should be low), ash value (total ash, acid-insoluble ash – high ash indicates soil/sand adulteration), and extractive value (how much dissolves in a specific solvent).
  4. Chemical Evaluation:
    • Simple chemical tests (phytochemical screening) to identify the class of active principles (see topic 6).
    • Quantitative chemical assays to determine the *amount* of active principle (e.g., total alkaloid content).
  5. Chromatographic & Spectroscopic Evaluation:
    • The most advanced method. Uses techniques like TLC and HPLC to create a chemical "fingerprint" of the drug.
    • This fingerprint can be compared to a standard, authentic sample to confirm identity and purity.

WHO guidelines for the assessment of herbal medicines

The World Health Organization (WHO) has established guidelines to help countries regulate and assess herbal medicines, ensuring they are safe, effective, and of good quality.

Key points from the guidelines include:

Phytochemical screening tests for secondary metabolites

These are simple, rapid, qualitative (test-tube) tests used in the lab to detect the *presence* of different classes of secondary metabolites in a plant extract.

Class of Compound Common Test(s) Positive Result
Alkaloids Mayer's Test
Dragendorff's Test
Wagner's Test
Creamy-white precipitate
Orange-red precipitate
Reddish-brown precipitate
Flavonoids Shinoda Test A piece of magnesium (Mg) ribbon and conc. HCl are added to the extract. A pink, red, or magenta color appears.
Steroids (Steroidal ring) Salkowski's Test
Liebermann-Burchard Test
(With conc. H2SO4) A red color appears in the lower acid layer.
(With acetic anhydride + conc. H2SO4) A color change, often to blue or green.
Triterpenoids Salkowski's Test
Liebermann-Burchard Test
(With conc. H2SO4) A yellow or golden-yellow color appears.
(With acetic anhydride + conc. H2SO4) A red or purple color.
Phenolic Compounds
(and Tannins)
Ferric Chloride (FeCl3) Test Addition of a few drops of FeCl3 solution to the extract gives an intense blue, green, or black color.