The Comparison of Hypersil GOLD with Other Commercially Available Fast LC Columns

The use of sub-2 µm particles as packing material has become increasingly popular for applications in High Throughput Screening assays and in Ultra High Pressure Liquid Chromatography (UHPLC). Sub-2 µm particle packed columns offer advantages over the more traditional columns packed with 3 and 5 µm particles through shorter analysis times, improvements in resolving power, sensitivity and efficiency. One of the major drawbacks of reducing the particle size is the increase in back pressure, which is a limiting factor of the HPLC instrument.



Where:
P is the pressure drop across the column (psi),
F is the mobile phase flow rate (ml/min),
u is the mobile phase viscosity (cP),
L is the column length (mm)
r is the column radius (mm)
dp is the particle diameter for the packing material of the column (?m)

Comparison Results

When choosing a sub-2 µm column it is therefore very important to consider whether the back pressure generated is going to exceed the 400 bar, making necessary the use of a UHPLC system.

Four columns were chosen for this study: column 1, 50 x 2.1mm packed with 2.7 um particles,
column 2, 50 x 2.1 mm packed with 1.7 um particles, column 3, 50 x 2.1 mm packed with 1.8 um particles and Hypersil GOLD, 50 x 2.1 mm packed with 1.9 um. The mobile phase used consisted of 35% MeCN and 65% water; a phenol standard was prepared in mobile phase and run on each column, at varying flow rates (from 0.02 to 1 ml/min).



Hypersil GOLD exhibited the highest theoretical plates for a given backpressure.