Analytical Biochemistry Notes
Subject Overview
Analytical Biochemistry covers the core instrumental techniques of the clinical biochemistry laboratory: spectrophotometry and colorimetry (Lambert's and Beer's laws), flame photometry and atomic absorption spectroscopy, the major chromatography types (paper, thin-layer, column, gas, ion-exchange, gel), and electrophoresis. A 4-credit core theory paper.
Subject at a glance
3
Units
19
Topics
4
Credits
3
Infographics
Unit-wise Syllabus
3 units — open any unit for detailed topic-wise notes, diagrams and important exam questions
- Theory of spectrophotometry and colorimetry
- Lambert's law and Beer's law
- applications
- general principles
- limitations
- instrumentation and applications of flame photometry
- atomic absorption spectroscopy principle and applications
Absorbance
A = ε c l
ε
Molar absorptivity
c
Concentration of the solution
l
Path length (usually 1 cm)
Unknown concentration
(A test / A standard) × concentration of standard
Study guide
This unit covers spectrophotometry and colorimetry, Lambert's and Beer's laws and applications, flame photometry principles, instrumentation and use and atomic absorption spectroscopy. Beer's law, absorbance proportional to concentration, is the idea behind most lab measurements.
- Introduction
- definition and types of chromatography — paper chromatography
- thin-layer chromatography (including HPTLC)
- column chromatography
- gas chromatography
- ion-exchange chromatography
- gel chromatography — principle
- instrumentation and applications of each
Paper / TLC
Partition and adsorption
Amino acids, sugars
Column
Adsorption or size
Purifying proteins
Gas
Volatility
Alcohol, drugs
Ion exchange
Charge
Haemoglobin variants, proteins
Gel filtration
Molecular size
Protein separation
Study guide
You study the principles, instruments and uses of paper, thin-layer (including HPTLC), column, gas, ion-exchange and gel chromatography. Learn what each technique separates and how.
- Introduction
- principle
- instrumentation
- types (paper and gel electrophoresis) and applications
- 1Apply serum to the gel or strip
- 2Apply voltage
Proteins move by charge
- 3Stain the bands
- 4Read the pattern
Albumin moves farthest
- 5Bands
Albumin, α1, α2, β, γ globulins
Study guide
You study electrophoresis: principle, instrumentation, types (paper and gel) and applications. Understand how charged molecules move in an electric field and how this separates proteins.
Why Analytical Biochemistry Matters
Photometry, chromatography and electrophoresis are the instruments of modern clinical and research laboratories, used for everything from blood glucose to protein analysis.
How to Prepare for Analytical Biochemistry
- Practise Beer-Lambert calculations.
- Prepare a comparison table of chromatography types.
- Learn the working and maintenance of each instrument.
Analytical Biochemistry: Frequently Asked Questions
What is Beer's law?
Beer's law states that the absorbance of light by a solution is directly proportional to the concentration of the absorbing substance.
What is chromatography?
Chromatography is a technique that separates the components of a mixture based on how they move between a stationary and a mobile phase.