1-1 Spectrophotometry And Electrophoresis of Haemoglobin

Question 1

Define spectrophotometry.

Answer 1

Method of analysing molecules on basis of their spectral properties.

Question 2

Define electrophoresis.

Answer 2

Method of analysing molecules on basis of charge (measure migration in electric field)

Question 3

Beer lambert law.

Answer 3

Absorbance of solution is proportional to concentration of the absorbing solute and distance (path length) travelled by light through sample.

Absorbance = E (constant) x concentration of absorbing substance (mol/L) x path length (cm)

Question 4

Spectrum of Hb

Single peak
Double

Answer 4

Deoxy

Oxy

Question 5

What is the difference between HbS and HbA?

Answer 5

HbS has 6th aa as valine (hydrophobic uncharged)

From glutamic acid (philic) negatively charged

HbA since more negatively charged moved further down to anode end in electrophoresis

Question 6

Haemoglobin types.

Oxy
Deoxy
Met

Answer 6

Hb and oxygen
Hb without oxygen
Hb with water or hydroxyl where Fe goes from 2+ to 3+

Attached to Fe