Beginners Guide to UPLC

Introduction

In 2004, separation science was revolutionized with the introduction of Ultra-Performance Liquid Chromatography [UPLC® Technology]. Significant advances in instrumentation and column technology were made to achieve dramatic increases in resolution, speed and sensitivity in liquid chromatography. For the first time, a holistic approach involving simultaneous innovations in particle technology and instrument design was endeavored to meet and overcome the challenges of the analytical laboratory. This was done in order to make analytical scientists more successful and businesses more profitable and productive.

For more than four decades, reducing stationary-phase particle size has been exploited to improve chromatographic separation efficiency. Until recently, LC technology had reached a plateau in which the benefits of reducing particle size could not be fully realized due to the negative influence of instrument band spreading and limited pressure range.

ACQUITY UPLC System

Figure 1: The ACQUITY UPLC® System.

The ACQUITY UPLC System [Figure 1] has removed those barriers, enabling columns packed with smaller particles [1.7 – 1.8 µm] to reach their theoretical performance, while precisely delivering mobile phase at pressures up to 1030 bar [15,000 psi], thus providing a new level of chromatographic performance.

UPLC Technology facilitates improvements of resolution, sensitivity and speed to be achieved, without compromise. Whether the separation goal is to achieve ultra-fast analysis, increase throughput while maintaining resolution, improving resolution while decreasing analysis time or achieve ultra-high resolution, the flexibility of the ACQUITY UPLC System enables method requirements to be met [Figure 2].

UPLC Technology versatality

Figure 2: Versatility of UPLC Technology – achieving resolution, sensitivity and speed.

This technology primer is designed to provide new, existing and potential UPLC users the ability to understand how UPLC Technology works, how to be successful with it, and how it can provide impactful results within their organization.

 

 

Bands, Peaks and Band Spreading A sample mixture is transferred from a sample vial into a moving fluidic stream [mobile phase]. The sample is then carried by the mobile phase to the head of a chromatographic column by a high pressure pump...
The Promise of Small Particles If we think about chromatographic resolution in the most basic sense, it is simply the width of two peaks relative to the distance between those peaks. If we can make those peaks narrower or further apart, we can improve resolution.
The Consequence of Improved Performance UPLC Technology facilitates improved chromatographic performance by minimizing system band spreading to produce more efficient separations in less time, thus achieving better data quality.
Improving Productivity with UPLC Technology By understanding the chromatographic principles outlined within this primer and how they are applied, it becomes apparent that more than just small particles and higher pressure must be considered to maximize separation performance.