HPLC systems consist of the following components:
1) Solvent Reservoir
Holds mobile phase solvents, multiple reservoirs for gradient systems.
2) Pump or Solvent Delivery System
Ensures consistent flow of mobile phase at high pressures.
3) Sample Injection System
Introduces the sample into the mobile phase stream; can be manual or automated.
4) Column
The core component where separation occurs.
Guard columns may be used to protect the main column from contamination.
5) Detector
Measures eluting compounds and generates signals. Common detectors include:
UV/Vis Absorbance Detector: Measures UV/visible light absorbance.
Fluorescence Detector: For fluorescent compounds or derivatized analytes.
Refractive Index Detector: Detects refractive index changes.
Mass Spectrometric Detector (LC-MS): Mass-based detection.
Electrochemical Detectors: Measures oxidation/reduction currents.
Data System (Recorder)
Captures detector outputs to create chromatograms. Modern systems interface with software for analysis.
Column Oven or Temperature Control
Maintains consistent temperature for reproducibility and optimal separation.
Additional Components:
Degasser: Removes dissolved gases to avoid bubbles.
Gradient Mixer: Mixes solvents in gradient systems.