UNDERSTAND THE PRINCIPAL AND

WORKING OF DIFFERENT EQUIPMENT:

1.HPLC : High performance liquid

chromatography

PRINCIPAL : High-performance liquid

chromatography (HPLC) is a separation technique
that can be used for the analysis of organic
molecules and ions. HPLC is based on mechanisms
of adsorption, partition and ion exchange,
depending on the type of stationary phase used.
HPLC involves a solid stationary phase, normally
packed inside a stainless-steel column, and a liquid
mobile phase. Separation of the components of a
solution results from the difference in the relative
distribution ratios of the solutes between the two
phases.
WORKING :
1. HPLC relies on pumps to pass a pressurized
liquid and a sample mixture through a column filled
with adsorbent, leading to the separation of the
sample components.
2.The active component of the column, the
adsorbent, is typically a granular material made of
solid particles (e.g., silica, polymers, etc.), 2–50
μm in size.
3.The components of the sample mixture are
separated from each other due to their different
degrees of interaction with the adsorbent particles.
The pressurized liquid is typically a mixture of
solvents (e.g., water, acetonitrile and/or methanol)
and is referred to as a "mobile phase".
4.Its composition and temperature play a major
role in the separation process by influencing the
interactions taking place between sample
components and adsorbent.
5.These interactions are physical in nature, such
as hydrophobic (dispersive), dipole–dipole and
ionic, most often a combination.