Instrumentation
General instrumentation for HPLC has following components.
i) One or more solvent reservoirs for the mobile phase.
ii) A pump to deliver the mobile phase with varying range of pressures up to several hundred atmospheres to achieve reasonable flow rates.
iii) Sampling valves or loops where the sample may be injected into the flowing mobile phase. Sample may be dissolved in mobile phase.
iv) Guard column or an on-line filter to prevent contamination of the main column.
v) A pressure gauge, inserted in front of the separation column, to measure column inlet pressure.
vi) Separation column containing packing to accomplish desired separation. These may be modified ion-exchange resin, silica gel, gel or a few other unique packing.
vii) A detector capable enough of measuring the solute concentrations.
viii) Recording and display device for plotting time vs peak intensity.
Besides, other electronic accessories for data manipulations are also required. These are schematically shown in Figure.
Figure: Schematic illustration of various components of HPLC instrument