Agilent Technologies has increased its focus on developing fast HPLC methods as the demand for rapid steroid detection screening methods continues to increase.
HPLC is a highly regarded analytical technique for pharmaceutical applications due to its unrivalled analytical validation characteristics; including accuracy, precision, limit of detection, specificity, linearity and range, and ruggedness.
Recent developments in HPLC column technology, such as sub-2µm totally porous and sub-3 µm superficially porous columns, have offered improvements in separation speed and performance through reduced analysis times and increased separation efficiency.
It is these improvements in column technology which mean that fast HPLC screening for the detection of steroids such as hydrocortisone, testosterone and progesterone can now be a focus.
Selectivity is a powerful tool to optimise separations in HPLC and can be manipulated to improve separations. As a result, the analysis of steroids can be dramatically improved by choosing the optimal HPLC column for the separation.
Selectivity can be increased by changing the mobile phase composition, column temperature and the composition of stationary phase.
The short column length and high efficiency of Agilent’s Poroshell 120 HPLC columns provide fast HPLC analysis times and rapid equilibration leading to fast investigations of selectivity.
A key feature of Poroshell 120 columns is their superficially porous microparticulate column packing. Poroshell 120 particles have a 1.7 µm solid silica core with a 0.5 µm porous outer layer.
This configuration delivers the performance advantages of sub-2 µm particles with the backpressure of a sub-3 µm particle.
With Poroshell 120 columns, it is possible to achieve 80-90% or more of the efficiency expected from a sub-2 µm Fast LC/UHPLC column – but at HPLC pressures (below 400 bar).
This ability to perform fast separations at low pressures can dramatically enhance productivity by allowing more samples to be run in less time – even using existing HPLC systems.
The Poroshell 120 family of columns offer the benefits of efficiency – more speed and more resolution – even on conventional HPLC instruments.
The improved quality of sub-2 µm particle columns in fast HPLC have less efficiency loss at high flow rates.
Fast HPLC screening using short columns packed with small particles and high flow rates is an effective solution to reduce analysis time.
Using alternative selectivity columns such as Poroshell 120 allows laboratories to achieve quicker turnaround and reporting times.
This allows laboratories to generate better chromatographic results quicker and so increase their throughput of samples. Use of HPLC selectivity columns resolves the compounds at better than baseline resolution.