HPLC is now a powerful analytical technique used for separating compounds soluble in solvents. It works by pumping a pressurized mobile phase through a column containing a stationary phase, which separates the compounds in a sample based on their interactions. The basic components of an HPLC system are a solvent reservoir, pump, injector, analytical column, detector, and recorder. HPLC can be used for quality control in pharmaceuticals and environmental and forensic applications.

1. High Performance Liquid
Chromatography(HPLC)
HPLC is now one of the most powerful tools in
analytical chemistry.
High Performance Liquid Chromatography (HPLC) is
an analytical technique used for the separation of
compounds soluble in a particular solvent.

2. HPLC Machine

3. Principle
High performance liquid chromatography (HPLC) is basically a
highly improved form of column liquid chromatography. It has
the ability to separate and identify the compounds that are
present in any sample.
Its basic component on the basis of their molecular structure
and molecular composition.
1. A stationary phase (a solid, or a liquid supported on a
solid): Its not moveable .
2.A mobile phase (a liquid or a gas): Gas or liquid that
carries the components of the mixture through the stationary
phase .

4. Components and function...
The basic components of an HPLC system
include:
• A solvent reservoir,
• Pump,
• Injector,
• Analytical column,
• Detector,
• Recorder/Data.
Types of HPLC:
1. Normal Phase HPLC.
2. Reverse Phase HPLC.
3. Size-exclusion HPLC.
4. Ion-Exchange HPLC.

5. HPLC system
•A solvent reservoir :To carry sample into the
column (mobile phase).
•Pump: (solvent delivery system or solvent
manager) To produce an appropriate pressure to
push solvent into the sample. Pumping solvent up to
a pressure of 4000 psi.
•An injector :It’s a sample manager that is able to
continuously flowing mobile phase stream that
carries into the HPLC column.
•The column : Mainly it is an analyzer . The
separation of the compound mixture takes place in
this column.

6. A detector : A detector is needed to see the
separated compounds from the HPLC column.
Data Collection Devices : The computer collect data
from detector and shows on the graph.

7. Application
1. Pharmaceuticals industry
• To control the drug stability.
• Pharmaceutical quality control.
2. Environmental Applications
-Detection of phenol & Mercury from drinking water.
3. Forensic test
- Determination of steroid in blood, urine & sweat.
4. Clinical test
Urine analysis, antibiotics analysis in blood.
5. Food and Flavor's
1. Measurement of Quality of soft drinks and water.
2. Sugar analysis in fruit juices.
3.Preservative analysis.