COLORIMETRY

Devesh Kumar Yadav

Hrishik Mukherjee
Sarthak Jain
INTRODUCTION:
 Photometry is the most common analytical technique used in the biochemical laboratory.
 It is designed to measure the intensity of a beam of light.
 Photometric principles are applied to the several kinds of analytical techniques:

(a) where absorbed or transmitted light is measured:

 Colorimetry, Spectrophotometry , Atomic absorption, and Turbidometry

(b) where emitted light is measured:

 Flame emission photometry
PRINCIPLE:
 Colored solutions have the property of absorbing certain wavelength of light when a
monochromatic light is passed through them.
 The amount of light absorbed or transmitted by a colored solution is in accordance with two
laws: – Beer’s law – Lambert’s law
MATERIALS REQUIRED:
METHOD:
BEER’S LAW:
LAMBERT’S LAW:
 Similar to Beer’s law, Lambert’s law states that:

  Equal parts of the same absorbing medium absorb equal fractions of the light that enters them. If in
traversing a path of length dx the intensity is reduced from I to I – dI then Lambert's law states that dI/I is
the same for all elementary paths of length dx. The relation is represented by the formula-

dI/I = -Kdx
BEER-LAMBERT’S LAW:
Beer-Lambert Law states that the amount of energy absorbed or transmitted by a solution is proportional
to the solution's molar absorptivity and the concentration of solute
PROCEDURE
 Let’s see
OBSERVATIONS:
1. PREPARATION OF SOLUTION OF DIFFERENT CONCENTRATIONS
OBSERVATIONS:
2. DETERMINATION OF ƛ MAX
OBSERVATIONS:
3. DETERMINATION OF UNKNOWN CONCENTRATION
GRAPH:
APPLICATIONS: