MBC 211: Physical and Analytical Biochemistry

Course Code and Title

• MBC 211
• Physical and Analytical Biochemistry

Course Contents

• Centrifugation:
  • Definition
  • Principle
  • Types of centrifuge
  • Types of centrifugation
  • Applications of centrifugation
  • Cell fractionation; methods of cell fractionation
• Structure, function, and fractionation of extracellular organelles
• Overview of instrumentation in medical biochemistry:
  • Autoclave
  • Hot plate
  • Pipette
  • Water distiller
  • Bunsen burner
  • Laboratory Refrigerator and Freezers
  • pH Meter
  • Balances
  • Dry Block Heater
  • Colorimeter
  • Dispensers
  • Flame Photometer
  • Fume Cupboard
  • Gas Analyzer
  • Gas Detector
  • Petri Dish
  • Measuring Cylinder
  • Photometer
  • Reagent Bottle
  • Homogenizer
  • Penetrometer
  • Spectrophotometer
  • Vortex etc.
• Human Specimen Collection and Handling
• Reference Ranges
• Quality Assurance and Quality Control in the Laboratory
• Laboratory Practical
• Centrifugation (lesson one 1a):
  • Definition
  • Principle
  • Types of centrifuge
  • Types of centrifugation
  • Applications of centrifugation
• Centrifugation technique: separation of substances using centrifugal force.
  • Particles are separated based on size, shape, density, viscosity of the medium and rotor speed

Principle of Centrifugation

• In a solution, higher-density particles sink and lower-density particles float.
• Centrifugal force separates particles based on density differences.
• Denser particles move away from the centre while less dense move towards.
• Tubes in a centrifuge are designed to allow larger particles to travel further outward from the middle.

Types of Centrifuge

• Low-Speed Centrifuge:
  • 4,000-5,000 rpm
  • Fixed angle
  • Swinging bucket
  • Used for sedimentation of red blood cells
• High-Speed Centrifuge:
  • 15,000-20,000 rpm
  • Used for sophisticated biochemical applications
  • Fixed angle
  • Swinging buckets
  • Vertical rotors

Types of Centrifugation

• Differential Pelleting (differential centrifugation):
  • Most common type
  • Homogenization of tissue samples
  • Spinning at various speeds to separate particles based on size and density
  • Creates pellet and supernatant
• Density Gradient Centrifugation:
  • Separates particles based on buoyant density in a density gradient.
• Rate-Zonal Density Gradient Centrifugation:
  • Separation based on sedimentation coefficient in a density gradient
• Isopycnic Centrifugation:
  • Separation based on density equilibrium in a gradient

Applications of Centrifugation

• Biological research
  • Cell separation
  • Protein purification
• Clinical diagnostics
  • Blood component separation
  • Urine analysis
• Food industry
  • Cream separation
  • Juice clarification
• Pharmaceuticals
  • Drug formulation
  • Vaccine production
• Environmental science
  • Water treatment
  • Soil analysis

Instrumentation and Equipment in Medical Biochemistry

• Spectrophotometers, high-performance liquid chromatography (HPLC), gas chromatography (GC), mass spectrometers, electrophoresis equipment, enzyme immunoassays, clinical chemistry analyzers, nuclear magnetic resonance (NMR) spectroscopy, polymerase chain reaction (PCR) machines, flow cytometers, microarray platforms, autoclave, hot plate, pipette, water distiller, bunsen burner, laboratory refrigerator and freezers, pH meter, balances, dry block heater, dispensers, colorimeters, reagent bottles, homogenizers, penetrometers, vortex mixers, and fume cupboards

Cell Fractionation Methods

• Differential Centrifugation
• Density Gradient Centrifugation
• Ultracentrifugation
• Filtration and size exclusion
• Magnetic bead separation

Human Specimen Collection and Handling

• Methods for collecting various types of specimens, including blood, urine, tissues, saliva, sputum, etc.
• Importance of labeling
• Storage conditions
• Transport protocols
• Safety procedures (PPE, disposal)

Types of Specimens

• Blood (venipuncture, capillary collection)
• Urine (random, midstream)
• Tissue (biopsies)
• Saliva
• Sputum
• Other specimens (stool, synovial fluid, CSF)

Quality Control and Assurance

• Standard Operating Procedures (SOPs)
• Training and competency
• Documentation
• Internal audits
• Management review
• Compliance with regulations (e.g., ISO 15189, CLIA)
• Control samples
• Calibration
• Proficiency testing
• Importance of quality control, and procedures to achieve high-quality outcomes.

Laboratory Practicals in Biochemistry (general)

• Importance of procedures, practical skills, safety, and data interpretation.
• Common lab techniques, including equipment, types of experiments, methods, calculations, data, analysis, and reporting.

Test your knowledge on the fundamentals of physical and analytical biochemistry including centrifugation techniques, instrumentation in medical biochemistry, and quality assurance in laboratory settings. This quiz covers essential concepts and applications relevant to biochemistry students.