DEPARTMENT OF MEDICAL BIOCHEMISTRY

FACULTY OF BASIC MEDICAL SCIENCES

COLLEGE OF HEALTH SCIENCES
UNIVERSITY OF UYO – UYO
COURSE CODE: MBC 211
COURSE TITLE: PHYSICAL AND ANALYTICAL BIOCHEMISTRY
SESSION: FIRST SEMESTER 2024/2025
CREDIT UNIT: (3 CH)
LECTURER: DR GRACE EKPO
COURSE CONTENTS:
 Centrifugation: Definition; Principle; Types of Centrifuge; Types of
Centrifugation; Applications of Centrifugation.
 Cell Fraction; 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 Analyser; 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.
LESSON ONE 1(a) Centrifugation: Definition; Principle; Types of
Centrifuge; Types of Centrifugation; Applications of Centrifugation.
CENTRIFUGATION
 Centrifugation is a technique of separating substances which involves the
application of centrifugal force.
 The particles are separated from a solution according to their size, shape,
density, the viscosity of the medium and rotor speed.

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 Principle of Centrifugation

 In a solution, particles whose density is higher than that of the solvent

sink (sediment), and particles that are lighter than it floats to the top.
 The greater the difference in density, the faster they move. If there is no
difference in density (isopycnic conditions), the particles stay steady.
 To take advantage of even tiny differences in density to separate various
particles in a solution, gravity can be replaced with the much more
powerful “centrifugal force” provided by a centrifuge.
 A centrifuge is a piece of equipment that puts an object in rotation around
a fixed axis (spins it in a circle), applying a potentially strong force
perpendicular to the axis of spin (outward).
 The centrifuge works using the sedimentation principle, where the
centripetal acceleration causes denser substances and particles to move
outward in the radial direction.
 At the same time, objects that are less dense are displaced and move to
the center.
 In a laboratory centrifuge that uses sample tubes, the radial acceleration
causes denser particles to settle to the bottom of the tube, while low-
density substances rise to the top.

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 TYPES OF CENTRIFUGE

LOW-SPEED CENTRIFUGE
Ts1) Most laboratories have a standard low-speed centrifuge used for
routine sedimentation of heavy particles
2) The low-speed centrifuge has a maximum speed of 4000-5000rpm
3) These instruments usually operate at room temperatures with no means
of temperature control.
4) Two types of rotors are used in it,
 Fixed angle
 Swinging bucket.
5) It is used for sedimentation of red blood cells until the particles are
tightly packed into a pellet and supernatant is separated by decantation.

HIGH-SPEED CENTRIFUGES
1. High-speed centrifuges are used in more sophisticated biochemical
applications, higher speeds and temperature control of the rotor chamber
are essential.
2. The high-speed centrifuge has a maximum speed of 15,000 – 20,000
RPM
3. The operator of this instrument can carefully control speed and
temperature which is required for sensitive biological samples.
4. Three types of rotors are available for high-speed centrifugation-
 Fixed angle

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 Swinging bucket
 Vertical rotors

ULTRACENTRIFUGES
1. It is the most sophisticated instrument.
2. Ultracentrifuge has a maximum speed of 65,000 RPM (100,000’s x g).
3. Intense heat is generated due to high speed thus the spinning chambers
must be refrigerated and kept at a high vacuum.
4. It is used for both preparative work and analytical work.

TYPES OF CENTRIFUGATION
1. Differential Pelleting (differential centrifugation)
 It is the most common type of centrifugation employed.
 Tissue such as the liver is homogenized at 32 degrees in a sucrose
solution that contains buffer.
 The homogenate is then placed in a centrifuge and spun at constant
centrifugal force at a constant temperature.
 After some time a sediment forms at the bottom of a centrifuge called
pellet and an overlying solution called supernatant.
 The overlying solution is then placed in another centrifuge tube which is
then rotated at higher speeds in progressing steps.
2. Density Gradient Centrifugation
 This type of centrifugation is mainly used to purify viruses, ribosomes,
membranes, etc.
 A sucrose density gradient is created by gently overlaying lower
concentrations of sucrose on higher concentrations in centrifuge tubes
 The particles of interest are placed on top of the gradient and centrifuge
in ultracentrifuges.
 The particles travel through the gradient until they reach a point at which
their density matches the density of surrounding sucrose.
 The fraction is removed and analyzed.
3. Rate-Zonal Density-Gradient Centrifugation
 Zonal centrifugation is also known as band or gradient centrifugation
 It relies on the concept of sedimentation coefficient (i.e. movement of
sediment through the liquid medium)
 In this technique, a density gradient is created in a test tube with sucrose
and high density at the bottom.
 The sample of protein is placed on the top of the gradient and then
centrifuged.
 With centrifugation, faster-sedimenting particles in sample move ahead

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 of slower ones i.e. sample separated as zones in the gradient.
 The protein sediment according to their sedimentation coefficient and the
fractions are collected by creating a hole at the bottom of the tube.

4. Isopynic Centrifugation
 The sample is loaded into the tube with the gradient-forming solution (on
top of or below pre-formed gradient, or mixed in with self-forming
gradient)
 The solution of the biological sample and cesium salt is uniformly
distributed in a centrifuge tube and rotated in an ultracentrifuge.
 Under the influence of centrifugal force, the cesium salts redistribute to
form a density gradient from top to bottom.
 Particles move to point where their buoyant density equals that part of
gradient and form bands. This is to say the sample molecules move to
the region where their density equals the density of gradient.
 It is a “true” equilibrium procedure since depends on bouyant densities,
not velocities
Eg: CsCl, NaI gradients for macromolecules and nucleotides – “self-
forming” gradients under centrifugal force.

APPLICATIONS OF CENTRIFUGATION
Centrifugation is a technique that uses centrifugal force to separate
components of a mixture based on their density, size, and shape. Here are
some key applications:
1. Biological Research:
o Cell Separation: Isolating different cell types from blood or
tissue samples.
o Protein Purification: Separating proteins based on size or
density, often used in biochemical research.
2. Clinical Diagnostics:
o Blood Component Separation: Isolating plasma or serum from
blood samples for testing.
o Urine Analysis: Concentrating and separating components for
diagnostic purposes.
3. Food Industry:
o Cream Separation: Separating cream from milk in dairy
processing.
o Juice Clarification: Removing pulp and sediment from fruit
juices.
4. Pharmaceuticals:
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 o Drug Formulation: Purifying active ingredients and removing
impurities.
o Vaccine Production: Concentrating viral particles or antigens.
5. Environmental Science:
o Water Treatment: Removing suspended solids and
contaminants from water samples.
o Soil Analysis: Separating soil particles for study of composition
and contamination.
6. Industrial Applications:
o Material Separation: In manufacturing processes, separating
materials based on density (e.g., in recycling).
o Chemical Reactions: Separating products from reaction
mixtures.
7. Nanotechnology:
o Nanoparticle Separation: Isolating nanoparticles for research
and application in various fields.
8. Genetics:
o DNA/RNA Purification: Isolating nucleic acids from cells or
tissues for genetic analysis.

LESSON ONE (1b): Cell Fraction; Methods of Cell Fractionation.

Cell fractionation is a laboratory technique used to separate cellular
components while preserving their individual functions. This process
allows researchers to study specific organelles or cellular structures in
detail. Here are some common methods of cell fractionation:
1. Differential Centrifugation
 Description: This method involves sequentially spinning a cell
homogenate at increasing speeds.
 Process:
o Cells are broken open (homogenized) to release their contents.
o The homogenate is centrifuged at low speeds to pellet the
largest components (e.g., nuclei).
o The supernatant is then centrifuged at higher speeds to
separate smaller organelles (e.g., mitochondria, microsomes).
 Applications: Commonly used to isolate organelles for biochemical
studies.
2. Density Gradient Centrifugation
 Description: This technique uses a density gradient (usually created
with a sugar or salt solution) to separate particles based on their
density.
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  Process:
o A homogenate is layered on top of a pre-formed gradient in a
centrifuge tube.
o When spun, organelles will migrate to their specific density
levels, forming distinct bands.
 Applications: Often used to purify specific organelles like ribosomes
or lipid membranes.

3. Isopycnic Centrifugation
 Description: A specialized form of density gradient centrifugation
where particles reach an equilibrium position at their buoyant density.
 Process:
o Similar to density gradient centrifugation, but the focus is on
maintaining the density of the gradient throughout the
centrifugation.
 Applications: Used for separating nucleic acids or specific protein
complexes.
4. Ultracentrifugation
 Description: Involves spinning samples at extremely high speeds (up
to 100,000 x g or more).
 Process:
o This method can effectively separate very small particles like
viruses or ribosomes.
 Applications: Used in molecular biology for studying macromolecules
and subcellular structures.
5. Filtration and Size Exclusion
 Description: Involves passing the homogenate through filters or gels
that separate components based on size.
 Process:
o Larger components are retained while smaller ones pass
through.
 Applications: Useful for separating whole cells from cell debris or for
isolating specific sizes of proteins.
6. Magnetic Bead Separation
 Description: Uses magnetic beads coated with antibodies to isolate
specific cell types or organelles.
 Process:
o Beads bind to target components, and a magnet is applied to
separate the bound components from the rest.
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  Applications: Frequently used in immunoprecipitation and cell sorting.
7. Gradient Centrifugation with Ultrafiltration
 Description: Combines gradient centrifugation with ultrafiltration
techniques.
 Process:
o After initial separation, ultrafiltration can refine the isolation of
specific organelles.
 Applications: Enhances purity and concentration of isolated
components.

LESSON TWO (2a): Structure, Function and Fractionation of Extracellular

Organelles
Extracellular organelles, such as exosomes and microvesicles, play critical
roles in intercellular communication and various physiological processes.
Here’s an overview of their structure, function, and methods for their
fractionation.
Structure
1. Exosomes:
o Size: Typically 30-150 nm in diameter.
o Origin: Formed within multivesicular bodies (MVBs) that bud
from the endosomal membrane.
o Membrane: Composed of a lipid bilayer enriched in specific
proteins, lipids, and RNA.
2. Microvesicles:
o Size: Larger than exosomes, ranging from 100-1,000 nm.
o Origin: Bud directly from the plasma membrane of cells.
o Membrane: Also composed of a lipid bilayer, containing
proteins and lipids that reflect the parent cell’s characteristics.
3. Apoptotic Bodies:
o Size: Generally larger than microvesicles, 1-5 µm in diameter.
o Origin: Formed during the process of apoptosis (programmed
cell death).
o Composition: Contain cellular debris, including organelles and
nuclear fragments.
Function
1. Exosomes:
o Cell Communication: Transfer proteins, lipids, and RNA
between cells, influencing recipient cell behavior.
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 o Immune Response: Modulate immune responses by presenting
antigens or delivering signaling molecules.
o Waste Management: Facilitate the removal of unwanted
materials from cells.
2. Microvesicles:
o Cell Signaling: Involved in local and systemic signaling
processes by transferring bioactive molecules.
o Homeostasis: Help maintain cellular homeostasis and respond
to stress by exchanging components.
o Tissue Repair: Play roles in wound healing and tissue
regeneration.
3. Apoptotic Bodies:
o Cell Clearance: Assist in the safe removal of dying cells by
phagocytes, preventing inflammation.
o Intercellular Signaling: May carry signals that influence
surrounding cells in tissue repair and regeneration.
Fractionation Methods
1. Differential Centrifugation:
o Process: A series of centrifugation steps at increasing speeds
to separate extracellular organelles based on size and density.
o Application: Initially pelleting larger debris, followed by the
isolation of microvesicles and exosomes.
2. Density Gradient Centrifugation:
o Process: Samples are layered over a gradient (e.g., sucrose or
iodixanol) and centrifuged, allowing separation based on
buoyant density.
o Application: Provides a more refined isolation of exosomes and
microvesicles, yielding high-purity fractions.
3. Filtration:
o Process: Utilizes filters with specific pore sizes to separate
extracellular organelles from larger particles.
o Application: Often used in conjunction with centrifugation to
enrich for exosomes and microvesicles.
4. Ultrafiltration:
o Process: Uses membranes that allow small molecules to pass
while retaining larger extracellular organelles.
o Application: Efficiently concentrates and purifies exosomes
from culture media or biological fluids.
5. Magnetic Bead Isolation:
o Process: Employs magnetic beads coated with antibodies

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 targeting specific surface proteins of exosomes or
microvesicles.
o Application: Allows for targeted isolation of specific subsets of
extracellular organelles based on their protein markers.
LESSON (TWO) 2b: Overview of Instrumentation in Medical Biochemistry
Instrumentation in medical biochemistry encompasses a variety of
tools and techniques used for analyzing biological samples, diagnosing
diseases, and monitoring health.
These instruments collectively facilitate the various analytical,
preparatory, and safety procedures essential in medical biochemistry,
ensuring high-quality research and accurate diagnostics.

Here’s an overview of key instruments and their applications:

1. Spectrophotometers
 Function: Measure the intensity of light absorbed by a sample at
specific wavelengths.
 Applications: Used for quantifying biomolecules (like proteins,
nucleic acids, and metabolites) and enzyme assays.
2. Chromatography Systems
 Types:
o High-Performance Liquid Chromatography (HPLC): Separates
and quantifies compounds in a mixture.
o Gas Chromatography (GC): Analyzes volatile compounds.
 Applications: Used for drug analysis, metabolic profiling, and
hormone measurement.
3. Mass Spectrometers
 Function: Measures the mass-to-charge ratio of ions to identify and
quantify molecules.
 Applications: Used for identifying proteins, metabolites, and drugs in
complex mixtures, as well as in proteomics and metabolomics
studies.
4. Electrophoresis Equipment
 Types:
o Agarose Gel Electrophoresis: Separates DNA or RNA
fragments.
o SDS-PAGE: Separates proteins based on size.
 Applications: Used in genetic analysis, protein purification, and

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 quality control.
5. Enzyme Immunoassays (EIAs) and ELISA Kits
 Function: Utilize antibodies to detect and quantify specific antigens
or antibodies in a sample.
 Applications: Commonly used for diagnosing infectious diseases,
measuring hormone levels, and screening for allergens.
6. Clinical Chemistry Analyzers
 Function: Automated systems that perform a wide range of
biochemical tests on blood and other body fluids.
 Applications: Used for routine tests like glucose, cholesterol, liver
function tests, and electrolyte panels.
7. Nuclear Magnetic Resonance (NMR) Spectroscopy
 Function: Analyzes molecular structure and dynamics based on the
magnetic properties of atomic nuclei.
 Applications: Used in metabolomics and structural biology to study
metabolites and macromolecules.

8. Polymerase Chain Reaction (PCR) Machines

 Function: Amplifies specific DNA sequences through thermal cycling.
 Applications: Widely used in genetic testing, pathogen detection, and
research applications.
9. Flow Cytometers
 Function: Analyze and sort cells based on their physical and chemical
characteristics.
 Applications: Used in immunology, cancer research, and diagnostics
for analyzing cell populations.
10. Microarray Platforms
 Function: Allow simultaneous analysis of thousands of genes or
proteins.
 Applications: Used in genomics and proteomics for gene expression
profiling and disease marker discovery.
11. Autoclave
 Purpose: Sterilizes equipment and media with high-pressure steam.
 Application: Critical for eliminating microorganisms before
experiments.
12. Hot Plate
 Purpose: Provides a controlled heat source.
 Application: Used for heating solutions and maintaining reaction
temperatures.
13. Pipette
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  Purpose: Accurately measures and transfers small liquid volumes.
 Application: Essential for sample preparation and assays.
14. Water Distiller
 Purpose: Produces purified water through distillation.
 Application: Supplies sterile water for experiments, reducing
contamination risks.
15. Bunsen Burner
 Purpose: Produces an open flame for heating and sterilization.
 Application: Commonly used for sterilizing tools and heating samples.
16. Laboratory Refrigerator and Freezers
 Purpose: Store samples at controlled low temperatures.
 Application: Refrigerators for short-term, freezers for long-term
preservation of biological materials.
17. pH Meter
 Purpose: Measures the acidity or alkalinity of a solution.
 Application: Essential for monitoring pH in various biochemical
assays and processes.

18. Balances
 Purpose: Accurately weighs samples and reagents.
 Application: Critical for preparing precise concentrations in
experiments.
19. Dry Block Heater
 Purpose: Provides uniform heat without water.
 Application: Used for incubating samples at specific temperatures.
20. Colorimeter
 Purpose: Measures the absorbance or transmittance of light in a
sample.
 Application: Useful for quantitative analysis of colored solutions.
21. Dispensers
 Purpose: Dispenses precise volumes of liquids.
 Application: Facilitates the quick and accurate addition of reagents.
22. Flame Photometer
 Purpose: Analyzes the concentration of certain metal ions by
measuring emitted light from a flame.
 Application: Commonly used for detecting sodium, potassium, and
lithium.
23. Fume Cupboard
 Purpose: Provides a ventilated workspace to safely handle hazardous

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 chemicals.
 Application: Essential for protecting users from toxic fumes and
vapors.
24. Gas Analyser
 Purpose: Measures concentrations of gases in a sample.
 Application: Used for respiratory gas analysis and environmental
monitoring.
25. Gas Detector
 Purpose: Detects the presence of harmful gases.
 Application: Ensures safety in the laboratory by monitoring for leaks.
26. Petri Dish
 Purpose: Cultivates microorganisms and cells.
 Application: Used for microbiological assays and cell culture
experiments.
27. Measuring Cylinder
 Purpose: Measures liquid volumes accurately.
 Application: Useful for preparing solutions in precise quantities.
28. Photometer
 Purpose: Measures light intensity or concentration of light-absorbing
substances.
 Application: Employed in various biochemical assays for
quantification.
29. Reagent Bottle
 Purpose: Stores chemicals and reagents safely.
 Application: Ensures proper labeling and storage conditions for
laboratory use.
30. Homogenizer
 Purpose: Mixes and breaks down samples to create uniform
suspensions.
 Application: Used for preparing tissue samples for analysis.
31. Penetrometer
 Purpose: Measures the consistency of semi-solid substances.
 Application: Commonly used in quality control of pharmaceutical
formulations.
32. Vortex Mixer
 Purpose: Mixes small volumes of liquid samples rapidly.
 Application: Used to ensure uniformity in samples and reagents.

LESSON (THREE) 3a: Human Specimen Collection and Handling

Human specimen collection and handling are critical steps in medical
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diagnostics and research, ensuring the accuracy and reliability of results.
Human specimen collection is a critical process in healthcare and
research, involving the gathering of biological samples for diagnostic
testing, monitoring, or research purposes.
Here’s an overview of key aspects:
Types of Specimens
1. Blood
o Methods: Venipuncture (drawing blood from a vein), capillary
collection (fingerstick).
o Uses: Blood tests for chemistry, hematology, blood cultures, etc.
2. Urine
o Types: Random, midstream clean catch, 24-hour collection.
o Uses: Urinalysis, culture tests, drug testing.
3. Tissue
o Methods: Biopsies (needle, excisional, or incisional).
o Uses: Pathology examinations, histology.
4. Saliva
o Methods: Swabs or collection containers.
o Uses: Hormone testing, DNA analysis.
5. Sputum
o Methods: Collected from the respiratory tract, often after deep
coughing.
o Uses: Microbiological cultures, cytology.
6. Other Specimens
o Stool: For gastrointestinal tests.
o Synovial Fluid: From joints for analysis.
o CSF (Cerebrospinal Fluid): Collected via lumbar puncture for
neurological testing.
Collection Procedures
1. Preparation
o Patient Identification: Confirm the patient’s identity and explain
the procedure.
o Informed Consent: Obtain consent from the patient for the
collection process.
2. Aseptic Technique
o Use sterile equipment and follow hygiene protocols to prevent
contamination.
3. Collection Timing
o Some tests require specific timing (e.g., fasting for glucose
tests, timed urine collections).
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 4. Volume
o Collect the correct volume to meet test requirements.
Insufficient amounts can lead to inaccurate results.
Handling and Storage
1. Labeling
o Clearly label all specimens with patient details (name, ID, date,
time of collection) and type of specimen.
2. Transport
o Use appropriate containers and packaging to prevent leakage
and contamination during transport to the laboratory.
3. Storage Conditions
o Follow specific guidelines for storage:
 Blood: Refrigerated or at room temperature, depending on
the test.
 Urine: Typically refrigerated; some tests may require
immediate processing.
 Tissue: Fixed in formalin or frozen based on the analysis
requirements.
Safety Considerations
1. Personal Protective Equipment (PPE)
o Use gloves, masks, and gowns as needed to protect against
exposure.
2. Biohazard Disposal
o Properly dispose of sharps and other biohazardous materials in
designated containers.

Quality Control
1. Training
o Ensure that all personnel involved in specimen collection are
adequately trained in techniques and protocols.
2. Documentation
o Maintain accurate records of all specimens collected, including
any issues encountered during the process.
LESSON (THREE) 3b: Reference Ranges
Reference ranges, also known as normal ranges, are important
benchmarks used in laboratory medicine to interpret test results.
They represent the values typically found in a healthy population and
serve as a guide for assessing individual patient results.

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 Here’s an overview:
1. Definition
Reference ranges indicate the range of values expected for a healthy
individual for a specific test.
These ranges are determined through statistical analysis of samples
from a healthy population.
2. Establishment of Reference Ranges
 Population Selection: Reference ranges are established using a
representative sample of a healthy population, considering age, sex,
ethnicity, and other factors.
 Statistical Methods: Typically, the range is defined as the mean ± 2
standard deviations (SD), encompassing about 95% of the healthy
population.
 Laboratory Variability: Reference ranges can vary between
laboratories due to differences in equipment, reagents, and
methodologies.
3. Factors Influencing Reference Ranges
 Demographics: Age, gender, race, and geographic location can all
affect normal values.
 Physiological Conditions: Factors like pregnancy, diet, and
medications can alter test results.
 Circadian Rhythms: Some values fluctuate throughout the day (e.g.,
cortisol levels).

4. Common Tests and Their Reference Ranges

 Blood Glucose: Normal fasting levels are typically between 70-100
mg/dL.
 Complete Blood Count (CBC):
o White Blood Cells (WBC): 4,500-11,000 cells/µL
o Hemoglobin:
 Men: 13.8-17.2 g/dL
 Women: 12.1-15.1 g/dL
 Lipid Profile:
o Total Cholesterol: <200 mg/dL
o LDL (bad cholesterol): <100 mg/dL

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 o HDL (good cholesterol): >40 mg/dL for men, >50 mg/dL for
women
 Liver Function Tests:
o Alanine Aminotransferase (ALT): 7-56 units/L
o Aspartate Aminotransferase (AST): 10-40 units/L
5. Interpretation of Test Results
 Above Normal Range: May indicate a potential health issue (e.g.,
diabetes, liver disease).
 Below Normal Range: Can also signify problems (e.g., anemia,
malnutrition).
 Contextual Interpretation: Always consider results in conjunction
with clinical symptoms and medical history.
6. Limitations of Reference Ranges
 Not Absolute: Individual results may fall outside the reference range
and still be clinically normal.
 Dynamic Nature: Health status can change, affecting the relevance of
the reference range over time.
7. Clinical Use
Reference ranges are essential for:
 Diagnosing conditions
 Monitoring disease progression
 Evaluating treatment efficacy

LESSON (FOUR)4a: Quality Assurance and Quality Control in the

Laboratory
Quality Assurance (QA) and Quality Control (QC) are essential
components of laboratory operations, ensuring the accuracy, reliability, and
consistency of test results.
Implementing robust QA and QC practices is essential for
laboratories to maintain high standards, comply with regulations, and
ensure the accuracy of diagnostic results.
This leads to improved patient outcomes and trust in laboratory
services.
Here’s an overview of both concepts:
Quality Assurance (QA)
Definition: QA encompasses all systematic activities designed to ensure
that the laboratory processes meet established standards and
requirements. It is focused on preventing errors and ensuring high-quality
outcomes.

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Key Components:
1. Standard Operating Procedures (SOPs):
o Development and implementation of written procedures for all
laboratory processes.
o Ensures consistency and compliance with protocols.
2. Training and Competency:
o Regular training for laboratory personnel to ensure they are
knowledgeable about procedures and equipment.
o Competency assessments to confirm staff can perform tests
accurately.
3. Documentation:
o Maintaining accurate records of all laboratory activities,
including training, test results, and equipment maintenance.
o Ensures traceability and accountability.
4. Internal Audits:
o Regular evaluations of laboratory processes to identify areas
for improvement and compliance with regulations.
5. Management Review:
o Periodic reviews by management to assess the effectiveness
of the QA program and implement necessary changes.
6. Compliance with Regulations:
o Adhering to local and international standards, such as ISO
15189 for medical laboratories or CLIA regulations.
Quality Control (QC)
Definition: QC involves the operational techniques and activities used to
monitor and control the quality of laboratory results. It focuses on
identifying defects in the process.
Key Components:
1. Control Samples:
o Use of control materials (known samples) to verify the accuracy
and precision of test results.
o Control limits are established, and results are monitored
against these limits.
2. Calibration:
o Regular calibration of laboratory instruments to ensure they
provide accurate measurements.
o Calibration schedules should be adhered to based on
manufacturer guidelines and regulatory requirements.
3. Proficiency Testing:
o Participation in external proficiency testing programs to

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 evaluate laboratory performance against peers.
o Helps identify areas needing improvement.
4. Statistical Process Control:
o Application of statistical methods to monitor and control
processes, helping to identify trends or shifts in data.
o Charts and graphs are often used to visualize performance.
5. Corrective Actions:
o Implementation of procedures for addressing any deviations
from expected results or quality standards.
o Documenting the root cause analysis and actions taken to
prevent recurrence.
Integration of QA and QC
 Continuous Improvement: Both QA and QC are part of a continuous
improvement cycle that aims to enhance laboratory performance.
 Risk Management: Identifying potential risks in laboratory processes
and implementing strategies to mitigate them.
 Patient Safety: Ultimately, QA and QC contribute to better patient
care by ensuring that test results are accurate and reliable.
LESSON (FOUR) 4b: Laboratory practicals
Laboratory practicals are hands-on sessions designed to teach
students and professionals how to perform various experiments and
procedures in a laboratory setting.
They provide valuable experience in applying theoretical knowledge
and developing practical skills.
Laboratory practicals are an essential part of scientific education,
providing the hands-on experience necessary to develop competent and
confident laboratory professionals.
Here’s an overview of what laboratory practicals typically involve:
1. Objectives of Laboratory Practicals
 Skill Development: Enhance technical skills related to
instrumentation, sample handling, and analytical techniques.
 Application of Theory: Bridge the gap between theoretical knowledge
and real-world applications.
 Problem-Solving: Develop critical thinking and troubleshooting skills
through practical experimentation.
 Data Analysis: Learn to collect, analyze, and interpret data accurately.
2. Components of a Laboratory Practical
 Preparation:
o Pre-Lab Assignments: Review background material, safety

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 protocols, and procedures before the lab session.
o Material and Equipment Setup: Ensure all necessary materials,
reagents, and equipment are prepared and functional.
 Conducting Experiments:
o Following Protocols: Adhere to standard operating procedures
(SOPs) for accuracy and safety.
o Data Collection: Systematically record observations and
measurements during the experiment.
 Safety Practices:
o Personal Protective Equipment (PPE): Use gloves, goggles, and
lab coats as appropriate.
o Emergency Procedures: Be familiar with the location of safety
equipment (e.g., eyewash stations, fire extinguishers) and
protocols for spills or accidents.
3. Types of Laboratory Practicals
 Biochemical Analysis: Techniques such as spectrophotometry,
chromatography, and electrophoresis.
 Microbiological Techniques: Culturing microorganisms, performing
sensitivity tests, and staining procedures.
 Chemical Synthesis: Conducting reactions and purifying products.
 Tissue Culture: Growing and manipulating cells in vitro for research
purposes.
4. Data Analysis and Interpretation
 Calculations: Performing necessary calculations (e.g., concentration,
dilution factors).
 Statistical Analysis: Applying statistical methods to assess data
reliability and significance.
 Reporting Results: Writing laboratory reports that summarize
objectives, methods, results, and conclusions.
5. Common Challenges
 Technical Difficulties: Equipment malfunction or unexpected results
can arise; problem-solving skills are crucial.
 Time Management: Balancing time effectively to complete all tasks
within the allocated session.
 Collaboration: Working effectively in teams, communicating findings,
and dividing tasks.

6. Evaluation and Feedback

 Peer Review: Engaging in discussions with peers about findings and
methodologies.
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  Instructor Feedback: Receiving constructive criticism on technique
and reporting to improve future performance.
7. Importance of Laboratory Practicals
 Real-World Relevance: Prepares students and professionals for
careers in healthcare, research, and industry.
 Critical Thinking: Encourages independent thinking and the
application of scientific principles.
 Innovation and Discovery: Fosters creativity and curiosity, leading to
new insights and advancements in science.

TEXTBOOKS:
(1) CROOK, MARTIN A. “CLINICAL BIOCHEMISTRY AND METABOLIC
MEDICINE” (YEAR OF PUBLICATION 2018) EIGHT EDITION.

(2) VASUDEVAN, P.M “TEXTBOOK OF BIOCHEMISTRY FOR MEDICAL

STUDENTS” (YEAR OF PUBLICATION 2017) SIX EDITION.

(3) CHATTERJEA, M.N “TEXTBOOK FOR MEDICAL BIOCHEMISTRY” (YEAR

OF PUBLICATION 2018) THREE EDITION.

(4) BHAGAVAN, N. “ESSENTIALS OF MEDICAL BIOCHEMISTRY” (YEAR OF

PUBLICATION 2019) SEVEN EDITION.

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