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MEDICAL INSTRUMENTS - PART II

CHAPTER 1: CHROMATOGRAPHY (Principles, Types & Parameters)

1. What is chromatography? Classify chromatography on the basis of equilibrium process.

Definition: Chromatography is a separation technique in which the components of a mixture are separated based on their different
distribution between two phases:
 Stationary phase
 Mobile phase
Classification of Chromatography (Based on Equilibrium Process)
Type Principle
Adsorption chromatography Separation based on differential adsorption of solute on a solid stationary phase
Partition chromatography Separation based on distribution of solute between two immiscible liquid phases
Ion-exchange chromatography Separation based on reversible exchange of ions
Gel filtration (size exclusion) Separation based on molecular size
Affinity chromatography Separation based on specific biological interaction
2. What is elution method? Explain it in terms of liquid chromatography.
Definition: Elution method is a chromatographic technique in Types of Elution
which the mobile phase continuously flows through the stationary 1. Isocratic elution – Mobile phase composition remains
phase to carry and separate analytes. constant
Elution in Liquid Chromatography 2. Gradient elution – Mobile phase composition
 A liquid solvent is used as mobile phase changes gradually
 Sample is injected at the top of the column Importance
 Components move at different speeds depending on:  Produces sharp, well-resolved peaks
 Affinity for stationary phase  Essential in HPLC
 Solubility in mobile phase
3. What is TLC? Describe its construction.
Definition: Thin Layer Chromatography (TLC) is a planar chromatographic technique where separation occurs on a thin layer of
adsorbent coated on a solid support.
Construction of TLC Plate Principle
A TLC plate consists of: Separation occurs due to differential adsorption of components on
 Support stationary phase.
 Glass, aluminum, or plastic sheet Applications
 Adsorbent layer  Drug analysis
 Silica gel (most common)  Amino acids separation
 Alumina or cellulose  Purity testing
 Thickness: ~0.25 mm
 Binder
 Calcium sulfate to hold adsorbent
4. What is paper chromatography? State the different types of paper chromatography.
Definition: Paper chromatography is a partition Types of Paper Chromatography
chromatography technique in which cellulose paper 1. Ascending chromatography
acts as the stationary phase and a liquid solvent acts as o Solvent moves upward by capillary action
the mobile phase. 2. Descending chromatography
 Stationary phase → Water molecules held in o Solvent moves downward due to gravity + capillary action
cellulose fibers 3. Ascending–descending chromatography
 Mobile phase → Organic solvent o Combination of both methods
Principle 4. Radial (circular) chromatography
Separation occurs due to different partition of solutes o Solvent moves radially from center
between stationary and mobile phases. 5. Two-dimensional chromatography
o Paper is developed in two directions with different solvents

5. What is retention factor (Rf) of a compound? How is it calculated?

Definition Characteristics
Retention factor (Rf) is the ratio of distance traveled by the solute to the  Rf value is constant for a given compound under
distance traveled by the solvent front. fixed conditions
Formula -  Rf value ranges between 0 and 1
�������� �������� �� �������� Significance
Rf =
�������� �������� �� ������� �����  Used for identification of compounds
 Helpful in purity testing
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6. What is retention time? Mention the factors influencing retention time.

Definition: Retention time (Rt) is the time taken by a solute to travel from Importance
the point of injection to the detector in column chromatography.  Used for qualitative identification
Factors Influencing Retention Time  Helps in peak separation
 Nature of stationary phase
 Composition of mobile phase
 Flow rate of mobile phase
 Column temperature
 Affinity of analyte for stationary phase
 Column length and particle size
7. Define chromatogram. Classify chromatogram.
Definition: A chromatogram is the final recorded output of a chromatographic separation showing separated components as
spots or peaks with respect to distance or time.
Classification of Chromatogram
A. Based on appearance
1. Planar chromatogram
o Spots on paper or TLC plate
2. Column chromatogram
o Peaks obtained in column techniques (GC, HPLC)
B. Based on detection
1. Qualitative chromatogram – shows presence/identity
2. Quantitative chromatogram – peak area/height indicates amount

8. Draw & label a typical chromatogram. Prove that Cs / Cm = K.

Typical Chromatogram (Labelled) Labels:
Detector response  X-axis → Time / Distance
↑  Y-axis → Detector response
| Peak B  Peaks → Separated compounds
| /\  Rt → Retention time
| / \
| Peak A \
| /\ \
| / \ \
|___ /____\______ _\____________→ Time / Distance
Rt1 Rt2
Proof of Cs / Cm = K
Let:
 Cs = concentration of solute in stationary phase
 Cm = concentration of solute in mobile phase
 K = distribution (partition) coefficient
At equilibrium, the solute distributes between two phases:
Concentration in stationary phase
K=Concentration in mobile phase
Cs
∴ K=
Cm​
Hence proved that Cs / Cm = K
Significance
 Basis of partition chromatography
 Determines retention behavior

9. If compounds travel 2.2 cm and solvent front travels 2.9 cm, calculate the retention factor.
Given:
 Distance traveled by compound = 2.2 cm
 Distance traveled by solvent front = 2.9 cm
Formula
�������� �������� �� ��������
Rf =
�������� �������� �� ������� �����
Calculation
�.�
Rf = �.� ​ = 0.7586 ≈ 0.76
Answer: Retention factor (Rf) = 0.76
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10. How can you prove that an analyte is in equilibrium between the two phases?
Concept: In chromatography, an analyte is continuously distributed ��
At equilibrium: K=
between the stationary phase and the mobile phase. This ��​
distribution follows a dynamic equilibrium. Where:
Proof of Equilibrium  Cs = concentration of analyte in stationary phase
 When a solute is introduced into a chromatographic system, it:  Cm = concentration of analyte in mobile phase
 Adsorbs onto the stationary phase  K = distribution (partition) coefficient (constant)
 Desorbs back into the mobile phase Since the ratio Cs/Cm remains constant at a given
 This process occurs repeatedly and rapidly temperature, the analyte is proven to be in equilibrium
between the two phases.
Conclusion (Viva line) - A constant distribution coefficient confirms equilibrium between stationary and mobile phases.
11. What is adsorbent? State the characteristics of adsorbent.
Definition: An adsorbent is a solid stationary Characteristics of a Good Adsorbent
phase used in chromatography on whose surface 1. Large surface area
the analyte molecules are adsorbed. 2. Uniform particle size
Examples of Adsorbents 3. Inert and chemically stable
 Silica gel 4. High adsorption capacity
 Alumina 5. Mechanical strength
 Activated charcoal 6. Reproducible adsorption behavior
Importance
 Controls retention time
 Affects resolution and separation efficiency

CHAPTER 2: HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC & DETECTORS)

1. Describe the major components of HPLC with block diagram.
Definition of HPLC: High Performance Liquid Chromatography (HPLC) is a high-resolution liquid chromatographic technique
used to separate, identify, and quantify components of a mixture under high pressure.
Major Components of HPLC Applications of HPLC
A. Solvent Reservoir  Drug analysis
 Contains mobile phase (single or mixed solvents)  Clinical biochemistry
B. Degasser  Toxicology
 Removes dissolved gases to prevent bubble formation  Hormone estimation
C. Pump
 Delivers mobile phase at high pressure (up to 400 bar)
 Maintains constant flow rate
D. Sample Injector
 Introduces a fixed volume of sample into the mobile phase
E. Guard Column
 Protects analytical column from contamination
F. Analytical Column
 Packed with stationary phase (silica-based particles)
 Actual separation occurs here
G. Detector
 Detects eluted analytes and converts them into electrical signals
H. Data Acquisition System
 Records, processes, and displays chromatogram
Block Diagram of HPLC

Solvent Reservoir → Degasser → Pump → Injector → Column → Detector → Recorder / Computer

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2. Define detector. Describe the criteria for choice of a detector.

Definition: A detector is a device that senses the presence of eluting analytes from the column and converts it into a measurable
electrical signal.
Criteria for Choice of a Good Detector Common HPLC Detectors
A. High sensitivity  UV–Visible detector
 Detects low concentration of analytes  Fluorescence detector
B. Good stability and reproducibility  Refractive index detector
 Produces consistent results  Electrochemical detector
C. Wide linear dynamic range
 Accurate response over broad concentration range
D. Low noise and drift
 Improves signal clarity
E. Non-destructive
 Allows sample recovery (if required)
F. Universal or selective response
 Should match analyte properties
G. Fast response time
 Suitable for narrow peaks in HPLC
3. Define detector sensitivity. Mention the factors affecting detector sensitivity.
Definition: Detector sensitivity is the ability of a detector to D. Noise level
respond to small changes in analyte concentration and  Lower noise improves sensitivity
produce a measurable signal. In simple words: The lower the E. Mobile phase composition
amount detected, the higher the sensitivity.  Should not absorb at detection wavelength
Factors Affecting Detector Sensitivity F. Temperature stability
A. Type of detector  Fluctuations reduce sensitivity
 Fluorescence > UV > Refractive index G. Electronic amplification
B. Wavelength selection (for UV detector)  Proper amplification enhances response
 Correct λmax increases sensitivity Importance
C. Flow cell volume  Detects trace-level analytes
 Smaller volume → higher sensitivity  Improves accuracy and precision

4. What is data acquisition system? Draw a block diagram of chromatographic data processing system.
Definition: A data acquisition system (DAS) is a computer- Functions of DAS
based system that:  Signal amplification
 Collects signals from detector  Analog-to-digital conversion
 Converts them into digital form  Peak integration
 Processes and stores chromatographic data  Quantitative analysis
 Storage and reporting
Block Diagram of Chromatographic Data Processing System

Detector Significance
↓  Produces accurate chromatograms
Signal Amplifier  Essential for quantitative HPLC analysis
↓
Analog to Digital Converter (ADC)
↓
Computer / Data Processor
↓
Display / Printer / Storage
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CHAPTER 3: LABORATORY SAFETY, BIOSAFETY & QUALITY CONTROL

1. What is cross infection? Classify laboratory hazards. How can you prevent them?
Definition of Cross Infection
Cross infection is the transfer of pathogenic microorganisms from one person, specimen, or surface to another, often within a
healthcare or laboratory setting.
Classification of Laboratory Hazards
Type Examples
Biological hazards (biohazards) Bacteria, viruses, fungi, blood-borne pathogens
Chemical hazards Acids, alkalis, solvents, toxic chemicals
Physical hazards Heat, sharp instruments, centrifuge accidents
Ergonomic hazards Poor posture, repetitive strain
Radiation hazards UV, X-rays, radioactive isotopes
Prevention of Cross Infection
 Follow standard precautions (gloves, masks, lab coats)
 Proper hand hygiene before and after lab work
 Use of sterile techniques
 Disinfection and sterilization of equipment and surfaces
 Proper waste disposal (sharps, biohazard waste)
 Avoid mouth pipetting and direct contact with specimens
2. Classify laboratory hazards. What is WHO safety code?
Classification of Laboratory Hazards
(See table above under Q1)
WHO Laboratory Safety Code
The WHO safety code provides guidelines to minimize laboratory hazards, including:
 Use of personal protective equipment (PPE)
 Safe handling and disposal of infectious and chemical materials
 Engineering controls (biosafety cabinets, fume hoods)
 Safe storage of reagents and samples
 Emergency procedures (spills, exposure, fire)
 Regular training and supervision
3. What are the factors influencing a test in a laboratory? How can you check bio-hazards?
Factors Influencing Laboratory Tests
 Pre-analytical factors
 Patient preparation, specimen collection, transport, and storage
 Analytical factors
 Equipment calibration, reagents, technique, operator skill
 Post-analytical factors
 Data interpretation, reporting errors
Checking for Biohazards
 Use biosafety cabinets for infectious samples
 Regular surface and instrument decontamination
 Proper waste segregation (biohazard containers)
 Use of autoclave / sterilization methods for contaminated materials
 Follow lab biosafety levels (BSL-1, 2, 3, 4) depending on risk

CHAPTER 4: STERILIZATION & DISINFECTION

1. Define sterilization. Write down the principle and uses of autoclave.
Definition of Sterilization: Sterilization is the process of complete destruction or removal of all forms of life, including: Bacteria,
Viruses, Fungi and Spores.
Autoclave
Principle of Autoclave
Autoclave works on the principle of moist heat sterilization under pressure.
 Saturated steam under pressure raises the temperature
 High temperature causes denaturation and coagulation of proteins of microorganisms, leading to death
Standard conditions:
 121°C at 15 psi for 15–20 minutes
Uses of Autoclave
 Sterilization of culture media  Glassware (some types)
 Surgical instruments  Bio-medical waste before disposal
 Dressings and linen
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2. What is autoclaving? Give the difference between sterilization and disinfection.

Autoclaving: Autoclaving is a method of sterilization using steam under pressure to kill all microorganisms including spores.
Difference Between Sterilization and Disinfection
Sterilization Disinfection
Destroys all microorganisms including spores Destroys most microorganisms (not spores)
Used for instruments and media Used for surfaces and skin
Example: Autoclave, hot air oven Example: Phenol, alcohol
Complete process Partial process
3. State the principle of hot air oven.
Principle of Hot Air Oven
Hot air oven works on the principle of dry heat sterilization.
 High temperature causes:
 Oxidation of cell components
 Protein denaturation
 Leads to death of microorganisms
Standard conditions:
 160°C for 2 hours or
 170°C for 1 hour
Uses of Hot Air Oven
 Glassware (petri dishes, pipettes)
 Metal instruments
 Powders and oils

CHAPTER 5: LABORATORY INSTRUMENTATION (PHYSICAL METHODS)

1. Illustrate the principle of centrifuge machine. Give its uses.
Principle of Centrifugation
Centrifugation works on the principle of sedimentation under centrifugal force.
 When a sample is rotated at high speed,
 Heavier particles move outward and settle at the bottom,
 Lighter particles remain at the top.
The force applied is called centrifugal force, which is much greater than gravity.
Uses of Centrifuge
 Separation of serum and plasma from blood
 Urine sediment examination
 Separation of cells and cellular components
 Clarification of samples
 Molecular biology and biochemistry applications
2. Classify pipette. Why mouth pipetting is prohibited? Why micropipette is used in the lab?
Classification of Pipettes
A. Based on calibration
1. Volumetric pipette – fixed volume
2. Graduated pipette – variable volume
B. Based on function
1. Transfer pipette
2. Pasteur pipette
C. Automatic pipettes
1. Micropipette
2. Electronic pipette
Why Mouth Pipetting is Prohibited
 Risk of chemical poisoning
 Risk of infection from biohazards
 Possibility of accidental ingestion
 Against WHO laboratory safety guidelines
Why Micropipette is Used in the Lab
 Accurate measurement of microliter volumes (µL)
 High precision and reproducibility
 Prevents contamination
 Essential for modern laboratory techniques (PCR, ELISA)
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CHAPTER 6: SPECTROSCOPY & OPTICAL METHODS

1. What is OD? Mention the estimation of HbA1C.
What is OD (Optical Density)?
Optical Density (OD) is a measure of how much light is absorbed by a substance when light passes through it.
 It is based on Beer–Lambert law
 OD is directly proportional to concentration of the substance

I /I
OD= log 0
Where:
 I0 = incident light
 I = transmitted light
Estimation of HbA1C
HbA1C is estimated by spectrophotometric or chromatographic methods.
Common laboratory methods:
1. HPLC (Gold standard)
2. Immunoturbidimetric method
3. Boronate affinity method
Principle (HPLC method):
 HbA1C is separated from other hemoglobin fractions
 Measured as a percentage of total hemoglobin
Clinical Significance of HbA1C
 Reflects average blood glucose over last 2–3 months
 Used for diagnosis and monitoring of diabetes
2. Illustrate Bohr’s effect. How can you determine HbA1C in your lab?
Bohr’s Effect
Definition
Bohr’s effect states that increase in CO₂ concentration or decrease in pH reduces hemoglobin’s affinity for oxygen, facilitating
oxygen release to tissues.

Illustration (Conceptual)
O₂ saturation
↑
| Normal pH
| /
| /
| / ↓ pH / ↑ CO₂
| /____ shifted right
|
+----------------------→ PO₂

Significance
 Helps oxygen delivery to metabolically active tissues
 Important in respiration and acid–base balance
Determination of HbA1C in Laboratory
By HPLC Method
1. Whole blood sample collected in EDTA
2. Hemoglobin fractions separated in HPLC column
3. HbA1C peak measured
4. Result expressed as % of total hemoglobin
Other Methods
 Immunoassay
 Enzymatic method
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CHAPTER 7: IMAGING TECHNOLOGY

1. What is MRI? How it works? Give its application.
Definition: MRI is a non-invasive imaging technique that uses a strong magnetic field and radiofrequency (RF) waves to produce
detailed images of soft tissues of the body.
How MRI Works (Principle) Applications of MRI
 The patient is placed in a strong magnetic field  Brain and spinal cord imaging
 Hydrogen nuclei (protons) align with the field  Tumor detection
 RF pulses disturb this alignment  Joint and soft tissue imaging
 When RF is stopped, protons release energy  Cardiac imaging
 The released signals are detected and converted into  Detection of ligament and muscle injury
images by a computer
2. Classify MRI. Mention the possible risks of MRI. How can they be minimized?
Classification of MRI Possible Risks of MRI
A. Based on magnet strength  Injury from metallic implants or objects
 Low-field MRI  Claustrophobia
 High-field MRI  Heating due to RF energy
B. Based on technique  Contrast-related reactions (rare)
 Functional MRI (fMRI) Minimization of MRI Risks
 Contrast-enhanced MRI  Proper patient screening for metal implants
 MR angiography (MRA)  Removal of all metallic objects
 Use of ear protection
 Monitoring patient during procedure
 Avoid contrast in high-risk patients
3. What is ultrasonogram? How it converts sound wave to image? Write down its application.
Definition: An ultrasonogram is an image produced by Applications of Ultrasonography
ultrasonography using high-frequency sound waves.  Pregnancy and fetal monitoring
Principle  Abdominal organ imaging (liver, kidney)
 A transducer emits ultrasound waves  Cardiac imaging (echocardiography)
 Sound waves reflect from tissues of different densities  Detection of cysts, tumors, stones
 Reflected waves (echoes) are received by the transducer  Guidance for biopsy
 Computer converts echoes into real-time images
CHAPTER 8: MOLECULAR BIOLOGY & GENETICS
1. What is DNA cloning? State five procedures of DNA cloning from any organism.
Definition
DNA cloning is the process of making multiple copies of a specific DNA fragment using a suitable host organism, typically bacteria.
It is used for gene analysis, protein production, and genetic engineering.
Five Procedures of DNA Cloning
1. Isolation of DNA
o Extract DNA from the source organism (e.g., human, bacteria).
2. Fragmentation
o Cut DNA into fragments using restriction enzymes.
3. Insertion into Vector
o Ligate the DNA fragment into a cloning vector (plasmid, bacteriophage, or cosmid).
4. Introduction into Host
o Introduce the recombinant DNA into a host cell (usually E. coli) via transformation.
5. Selection and Screening
o Select host cells that contain the recombinant DNA using antibiotic resistance or marker genes.
o Confirm using colony PCR, restriction analysis, or sequencing.
2. What is PCR? Give Illustration, Steps, and Advantages
Definition: PCR (Polymerase Chain Reaction) is a technique to amplify a specific DNA segment rapidly in vitro using DNA
polymerase.
Illustration (Conceptual Diagram)
DNA template
↓
Denaturation (95°C) → strands separate
↓
Annealing (50–65°C) → primers bind
↓
Extension (72°C) → DNA polymerase synthesizes new strand
↓
Cycle repeats → Exponential amplification of target DNA
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Steps of PCR Advantages of PCR

A. Denaturation  Rapid amplification of DNA
 Heat to 94–95°C to separate DNA strands.  Can detect tiny amounts of DNA
B. Annealing  High specificity due to primer design
 Cool to 50–65°C for primers to bind to complementary  Useful in diagnosis of genetic diseases,
sequences. infections, and forensic analysis
C. Extension  Enables gene cloning and sequencing
 Heat to 72°C; DNA polymerase synthesizes new DNA.
D. Repeat
 Cycle repeated 25–35 times → exponential amplification.
3. What do you mean by Chromosomal Study? Importance of Chromosomal Study
Definition: Chromosomal study (also called karyotyping) is the analysis of chromosomes in a cell to detect structural or numerical
abnormalities.
 Chromosomes are visualized during metaphase using staining techniques (e.g., G-banding).
 Helps in identifying genetic disorders, congenital anomalies, and cancer-related changes.
Importance of Chromosomal Study
A. Detection of genetic disorders
 e.g., Down syndrome (Trisomy 21), Turner syndrome (45,X)
B. Prenatal diagnosis
 Amniocentesis and chorionic villus sampling
C. Cancer diagnosis and prognosis
 Chromosomal translocations in leukemia
D. Infertility investigations
 Structural abnormalities like inversions or translocations
E. Research
 Understanding gene function and chromosomal behavior

4. What is Edman Reaction? Write Down the Steps of Post-Translational DNA-Sequencing Events on Ribosome
Edman Reaction
Definition:
Edman degradation is a method to determine the N-terminal amino acid sequence of a protein.
 Sequential removal of one amino acid at a time from the N-terminus.
 Uses phenylisothiocyanate (PITC) reagent.
Steps of Edman Reaction
A. Reaction with PITC
 N-terminal amino acid reacts with phenylisothiocyanate under alkaline conditions.
B. Cleavage
 The N-terminal amino acid is cleaved from the peptide as a phenylthiohydantoin (PTH) derivative.
C. Identification
 PTH-amino acid is identified using chromatography.
D. Repeat Cycle
 Process is repeated to sequence the next amino acid.
Post-Translational DNA-Sequencing Events on Ribosome (Simplified Steps)
 Transcription – DNA → mRNA in the nucleus
 mRNA processing – 5’ capping, splicing, 3’ poly-A tail
 Translation initiation – mRNA binds to ribosome; first tRNA binds
 Elongation – Sequential addition of amino acids to growing polypeptide
 Termination – Stop codon recognized; polypeptide released
 Post-translational modifications – Folding, phosphorylation, glycosylation, etc.

CHAPTER 9: CLINICAL BIOCHEMISTRY

1. Why blood gases analysis is necessary? Prepare a table for ABG with clinical significance.
Why Blood Gas Analysis (ABG) is Necessary ABG Parameters with Normal Values & Clinical Significance
Arterial Blood Gas (ABG) analysis is necessary to: Parameter Normal Value Clinical Significance
 Assess oxygenation status pH 7.35–7.45 Acidosis / alkalosis
 Evaluate acid–base balance PaO₂ 80–100 mmHg Oxygenation status
 Monitor respiratory function PaCO₂ 35–45 mmHg Respiratory function
 Diagnose metabolic and respiratory disorders HCO₃⁻ 22–26 mmol/L Metabolic component
 Monitor critically ill patients (ICU, ventilated patients) O₂ Saturation 95–100% Tissue oxygen delivery
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2. Explain “Salting Out Effect”. Give the difference between fractionation and dialysis of protein. Mention the significance of
SOE.
Salting Out Effect (SOE)
Definition: Salting out effect is the precipitation of proteins from solution by adding high concentration of neutral salts (e.g.,
ammonium sulfate).
Principle
 Salt ions attract water molecules
 Less water is available to solubilize proteins
 Protein molecules aggregate and precipitate
Common Salt Used
 Ammonium sulfate
Difference between Fractionation and Dialysis of Protein
Fractionation Dialysis
Used to separate proteins from a mixture Used to remove small molecules (salts, ions) from protein
solution
Based on difference in solubility of proteins Based on difference in molecular size
Commonly uses salting-out effect (e.g., ammonium sulfate) Uses a semi-permeable membrane
Protein precipitates during the process Protein does not precipitate
Helps in protein purification and isolation Helps in desalting and buffer exchange
Example: Separation of albumin and globulin Example: Removal of ammonium sulfate after fractionation
Significance of Salting Out Effect
 Protein purification
 Separation of plasma proteins
 Enzyme isolation
 Removal of unwanted proteins

CHAPTER 10: RADIOACTIVITY & RADIATION CHEMISTRY

1. What is radiation process? Write down the clinical application of radioactive rays.
Radiation Process: Radiation is the emission and Clinical Applications of Radioactive Rays
propagation of energy in the form of particles or A. Cancer radiotherapy
electromagnetic waves from unstable atomic nuclei.  Gamma rays (Co-60) used to destroy tumor cells
Types of radiation: B. Nuclear medicine imaging
 Alpha (α) rays – helium nuclei  PET and SPECT scans for organ function assessment
 Beta (β) rays – electrons or positrons C. Thyroid disorders
 Gamma (γ) rays – high-energy electromagnetic waves  I-131 for diagnosis and treatment of hyperthyroidism
D. Tracer studies
 Study metabolism, blood flow, and organ function
E. Sterilization
 Gamma radiation for medical equipment sterilization

2. What is Grignard reaction? State the chemical properties of radioisotopes. How deduction of radioactivity is done?
Grignard Reaction
Definition: Grignard reaction involves the reaction of an organomagnesium halide (RMgX) with compounds containing carbonyl
groups, forming alcohols.
General reaction:
RMgX + R′CHO → R′CH(OH)R
Uses:
 Synthesis of alcohols
 Formation of carbon-carbon bonds
Chemical Properties of Radioisotopes
 Same chemical behavior as stable isotopes
 Participate in same chemical reactions
 Can form identical compounds
 Differ only in nuclear stability
Deduction (Detection) of Radioactivity
Radioactivity is detected by measuring emitted radiation using:
 Geiger-Müller counter
 Scintillation counter
 Ionization chamber
 Film badge / TLD (dosimeter)
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CHAPTER 11: THERMODYNAMICS (PHYSICAL CHEMISTRY)

1. Elucidate entropy and enthalpy. Establish the relationship between them.
Enthalpy (H): Enthalpy is the total heat content of a system at Relationship between Enthalpy and Entropy
constant pressure. The relationship is given by Gibbs free energy equation:
H=U+PV ΔG = ΔH−TΔS
Where: Where:
 U = internal energy  ΔG = Gibbs free energy change
 P = pressure  ΔH = Enthalpy change
 V = volume  T = Absolute temperature
Change in enthalpy (ΔH) represents heat absorbed or released  ΔS = Entropy change
during a reaction.
Entropy (S): Entropy is a measure of randomness or disorder of a For a reaction to be spontaneous, ΔG <0
system.
 Higher disorder → higher entropy
 Spontaneous reactions increase entropy NOTE: Delta - Δ
CHAPTER 12: TUMOR MARKERS & ONCOLOGY
1. What do you mean by tumor marker? Enumerate the tumor markers with their indications.
Definition: A tumor marker is a substance produced by cancer cells or by the body in response to cancer, which can be measured in
blood, urine, or tissue.
Common Tumor Markers and Their Indications Uses of Tumor Markers
Tumor Marker Indication  Cancer diagnosis (supportive)
AFP Liver cancer, germ cell tumor  Monitoring treatment response
CEA Colorectal cancer  Detecting recurrence
PSA Prostate cancer  Prognosis
CA-125 Ovarian cancer
CA 19-9 Pancreatic cancer
CA 15-3 Breast cancer
β-hCG Testicular cancer, trophoblastic disease

CHAPTER 13: ANDROLOGY / SEMEN ANALYSIS

1. State the composition, characteristics and importance of semen.
Composition of Semen Importance of Semen
Component Source  Essential for fertilization
Spermatozoa Testes  Provides nutrition and transport for sperm
Seminal fluid Seminal vesicles  Used to assess male fertility
Enzymes Prostate gland  Important in medico-legal cases
Fructose Seminal vesicles
Alkaline fluid Prostate
Characteristics of Normal Semen
Parameter Normal Value
Volume 2–6 mL
Color Grayish white
pH 7.2–8.0
Sperm count ≥ 15 million/mL
Motility ≥ 40%
Morphology ≥ 4% normal forms

Extra Q: What do you mean by LASER? Enumerate its properties and latest medical applications
Definition of LASER: LASER stands for Light Amplification Latest Medical Applications of LASER
by Stimulated Emission of Radiation. A. Surgery
 It is a device that produces a highly focused,  Eye surgery: LASIK, cataract removal
monochromatic, and coherent beam of light.  Soft tissue surgery, tumor ablation
 LASER light is directional, intense, and of single B. Dermatology
wavelength.  Removal of tattoos, scars, and wrinkles
Properties of LASER  Hair removal
A. Monochromatic – Emits light of a single wavelength. C. Oncology
B. Coherent – Light waves are in phase in time and space.  Laser therapy for tumor coagulation and ablation
C. Directional – Travels in a narrow, collimated beam. D. Dentistry
D. High intensity – Very bright and powerful beam.  Teeth whitening, gum surgery, cavity treatment
E. Stimulated emission – Light is produced by stimulated E. Imaging & Diagnosis
emission of photons.  Confocal microscopy, optical coherence tomography
F. Photodynamic Therapy (PDT)
 Cancer treatment using photosensitizing drugs and laser light