UHPLC

UHPLC (Shimadzu, Corp., Japan)

The fundamental principle of High-Performance Liquid Chromatography (HPLC) is the separation of compounds in a mixture based on their different interactions with a stationary phase and a mobile phase. The mobile phase (a liquid solvent) carries the sample through a column packed with a stationary phase. Components that interact more strongly with the stationary phase move slower, while those with weaker interactions move faster, leading to their separation and subsequent detection as they exit the column. The fundamental principle of UHPLC is that it improves upon traditional HPLC by using highly efficient columns packed with sub-2-µm particles, which requires high-pressure systems to achieve higher linear velocities. This approach, guided by the Van Deemter equation, optimizes the correlation between flow rate and plate height (column efficiency), leading to enhanced separation speed, resolution, and sensitivity compared to HPLC.

Key components

Mobile Phase: A liquid solvent or a mixture of solvents that continuously flows through the column, carrying the sample.

Stationary Phase: A material (often solid particles) packed inside the column that interacts with the sample components.

Column: A cylindrical tube containing the stationary phase, where the separation process takes place.

Pump: Delivers the mobile phase at high pressure to force the sample through the column.

Injector: Introduces the sample mixture into the mobile phase stream.

Detector: Identifies and measures the separated components as they elute from the column, generating a signal that is plotted as a chromatogram.

Application

Amino acid analysis, vitamin (A and E) analysis.

Sample type: Meat, milk, feed (fresh/dry)

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