HPLC with UV detector

Description

High-Performance Liquid Chromatography (HPLC) with UV Detector
High-Performance Liquid Chromatography (HPLC) with a UV Detector is a sophisticated analytical technique widely used in chemistry, biochemistry, pharmaceuticals, environmental analysis, and food science to separate, identify, and quantify components in complex mixtures.

HPLC works by passing a liquid sample mixture through a column packed with a stationary phase under high pressure. Different components in the mixture interact variably with the stationary phase and are thus separated based on their chemical properties. The separated compounds then pass through a UV (Ultraviolet) Detector, which measures their absorbance of ultraviolet light, enabling detection and quantification.

Working Principle
Separation Process
The sample mixture is injected into a pressurized liquid mobile phase, which carries it through a chromatographic column filled with a stationary phase (usually silica-based). Components in the mixture separate due to differences in polarity, size, or affinity to the stationary phase.

Detection by UV Detector
As components elute from the column at different times (retention times), they pass through the UV detector. The UV detector shines ultraviolet light (commonly between 190–400 nm) through the eluent. Compounds absorbing UV light reduce its intensity, and this absorbance is converted into an electrical signal. The detector records these signals, producing chromatograms with peaks corresponding to each compound.

Key Components
Solvent Reservoirs
Contain the mobile phase solvents.

Pump
Delivers the mobile phase at high pressure (up to 6000 psi or more).

Injector
Introduces the liquid sample into the mobile phase stream.

Chromatographic Column
Packed with stationary phase material that separates sample components.

UV Detector
Measures absorbance of UV light by eluted compounds for identification and quantification.

Data System
Processes the electrical signals into chromatograms for analysis.