High-performance liquid chromatography (HPLC) is an analytical technique used to separate, identify, and quantify components in a mixture by passing a pressurized liquid solvent through a column filled with adsorbent material. Each component interacts differently with the adsorbent, leading to their separation based on varying flow rates. Factors influencing the process include the composition and temperature of the mobile phase, internal diameter of the tubing, particle size, and pump pressure.

1. CLINICAL BIOCHEMISTRY
PRESENTATION
HPLC
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
MADEBY: “RISHABHSHARMA”
B.SC CLINICAL RESEARCH

2.  High-performance liquid
chromatography (HPLC) is a technique
in analytical chemistry used to separate,
identify, and quantify each component in a
mixture.
 It relies on pumps to pass a pressurized
liquid solvent containing the sample
mixture through a column filled with a
solid adsorbent material.

3. Each component in the sample interacts slightly differently with the
adsorbent material, causing different flow rates for the different
components and leading to the separation of the components

4.  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.
 The active component of the column, the
adsorbent, is typically a granular material made
of solid particles 2–50 μm in size.
 The components of the sample mixture
are separated from each other due to
their different degrees of interaction
with the adsorbent particles.

5. The pressurized liquid is typically a mixture of
solvents (e.g., water, acetonitrile and/or methanol)
and is referred to as a "mobile phase".
 Its composition and temperature play a major
role in the separation process by influencing
the interactions taking place between sample
components and adsorbent.

6. Types
Partition chromatography
Normal–phase chromatography
Displacement chromatography
Reversed-phase chromatography (RPC)
Size-exclusion chromatography
Ion-exchange chromatography
Bioaffinity chromatography

7.  FACTORS AFFECTING CHROMATOGRAPHY
 Internal diameter-Tubing on a system,
 Particle size
 Pore size
 Pump pressure

11. REFERENCES:-
HTTPS://EN.WIKIPEDIA.ORG/WIKI/HIGH-
PERFORMANCE_LIQUID_CHROMATOGRAPHY
HTTPS://WWW.GOOGLE.COM/SEARCH?TBM=ISCH&Q=HPLC+I
MAGES&CHIPS=Q:HPLC+IMAGES,G_1:PRINCIPLE:YNABBVPMP5
G%3D&USG=AI4_-
KSXKUBIWCHZRRI4RWPN305EJ63DJQ&SA=X&VED=0A
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