Question

Discuss the principles and application of analytical centrifugation.

Answer 1

Analytical centrifugation: It is a separation method in which centrifugal force is applied so that different components of the sample are separated by density differences of components i.e., sedimentation of heterogenous components by using centrifugal force in a centrifuge.

Principles of analytical centrifugation:

Analytical centrifugation abides by the following principles.

1. The denser a component is, the faster it sediments in the centrifugal field.
2. The more massive a component is, the faster it moves in a centrifugal field.
3. The denser the biological buffer system is, the slower the particle moves in a centrifugalfield.
4. The greater the frictional coefficient is, the slower a particle moves.
5. The greater the centrifugal force is, the faster the particle sediments.
6. The sedimentation rate of a given particle will be zero when the density of the particle and the surrounding medium is equal.

Applications of analytical centrifugation:

This technique is employed to characterize the mass, size, shape, and association properties of macromolecules in solution. Some applications are listed below

1. It can be used to determine the sample purity.
2. Determine the relative molecular mass.
3. Estimation of purity of a macromolecule.
4. Determination of conformational changes in molecules like DNA and proteins on denaturation.