Veterinary Clinical Pathology - 4th Level

Veterinary Clinical Pathology - 4th Level

• Course Title: Clinical Pathology (General Program)
• Instructor: Dr. Jihad Al-Saied
• Section: Second Section

Atomic Absorption

• Use: Measuring heavy metals
• Principle: Atomization (dissociation from chemical bonds, placing in unexcited state). Each element absorbs characteristic wavelength (W.L.) in the ground state.
• Process: Heavy metals separated from their chemical bonds then placed in the unexcited state.  Each element absorbs a unique wavelength.

Chromatography

• Use: Separating mixtures into components for identification, purification, and quantification
• Types:
  • Plane
  • Column
• Column Types:
  • Liquid-liquid chromatography
  • Ion exchange chromatography
  • Steric (size) exclusion chromatography
  • HPLC
  • Gas chromatography

1- Liquid-liquid Chromatography

• Separation: Based on solubility

2- Ion Exchange Chromatography

• Separation: Based on anion exchange to separate unwanted substances

3- Steric (size) Exclusion Chromatography

• Use (Sephadex): Smaller molecules diffuse into the gel column, while larger ones pass rapidly, appearing first in the fluid discharge. Smaller molecules will be delayed in the column.

4- HPLC

• Method: Controlled pore size with pressure between 500-5000 pounds per square inch.

5- Gas Chromatography

• Characteristics: Rapid, sensitive, accurate
• Use: Separating and measuring nanogram and picogram amounts of volatile substances (steroids, lipids, drugs, blood alcohol, other toxic substances).

Electrochemistry

• Method: Measures ions in solution by their electrical properties.
• Techniques: Ion-selective electrodes (ISEs)
• Selective membrane: Measures single ions (Na+, K+, Cl-, Ca2+, HCO3+)
• Ion Contact with Membrane: Penetration changes potential measurements as voltage changes due to ionic activity.

Electrophoresis

• Use: Fractionating serum proteins
• Method: Migration of proteins in an electrical field
• Factors: Direction and rate of migration based on protein charge and size
• Buffer pH (8.6): Negative charge applied to serum proteins.

Electrophoresis - Molecules in Motion

• Migration Direction: Molecules move towards the opposite charged electrode.
  • Anions (-ve charge) towards the positive (anode) electrode.
  • Cations (+ve charge) towards the negative (cathode) electrode.
• Albumin: Strongest negative charge, migrates fastest toward the anode.
• Globulin: Weaker negative charge, migrates slowly toward the anode.
• Migration Pattern: Forms bands (use of media support).

This quiz covers key concepts in Clinical Pathology focusing on Atomic Absorption and Chromatography methods. It emphasizes the techniques used for measuring heavy metals and separating mixtures for analysis. Ideal for students in the Clinical Pathology program.