Beginner's Guide to UPLC

Beginner's Guide to UPLC

Introduction

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.

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].

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.

In this Primer

Introduction

Bands, Peaks and Band Spreading

The Promise of Small Particles

The Consequence of Improved Performance

Improving Productivity with UPLC Technology

 

Related

ACQUITY Premier System
Experience the ultimate in sensitivity, reproducibility, and speed. The ACQUITY Premier LC-MS provides the best sensitivity and peak shape for analytical separations.

ACQUITY UPLC I-Class PLUS System
The ACQUITY UPLC I-Class PLUS System offers dramatically decreased dispersion for the highest resolution, rapid injection cycles and sample throughput with reduced carryover, making it a superior inlet to any mass spectrometer.

ACQUITY UPLC H-Class PLUS System
The ACQUITY UPLC H-Class PLUS System delivers the flexibility and precision of quaternary or binary solvent blending with the advanced performance expected of UPLC separations: high resolution, sensitivity and improved throughput.

UPLC Columns
Get faster separations with higher resolution and sensitivities using sub-2-µm particles at 1241 bar pressures with UPLC Columns.
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Ultra Performance Liquid Chromatography (UPLC) represents an advanced chromatographic method tailored for rapid and high-resolution separation of chemical compounds. It evolves from the principles of High-Performance Liquid Chromatography (HPLC) but distinguishes itself by utilizing columns with notably smaller particle sizes and higher pressures, resulting in enhanced efficiency and accelerated analyses.

Here is an introductory overview of UPLC:

Principle: UPLC adheres to the fundamental principles of traditional liquid chromatography, involving the separation of sample components based on their interactions with a stationary and mobile phase. The crucial distinction lies in the utilization of columns with significantly smaller particle sizes, enhancing separation efficiency.

Benefits

- Speed: UPLC offers faster analysis times than HPLC, thanks to reduced particle size, facilitating quicker and more efficient separations.

- Resolution: Smaller particle sizes contribute to higher resolution, enabling the separation of closely eluting peaks and providing sharper chromatographic peaks.

- Sensitivity: UPLC often enhances sensitivity, particularly advantageous for trace-level analyses.

- Sample Throughput: Faster run times and improved efficiency increase the number of processed samples within a given timeframe.

Instrumentation: UPLC systems include pumps, detectors, and columns designed for higher pressures associated with smaller particle sizes. These systems seamlessly integrate with other analytical instruments for comprehensive data acquisition.

Applications: UPLC finds applications across various fields, including pharmaceuticals, environmental analysis, and food and beverage testing, where rapid and precise separations are crucial.

Beginner's Tips

- Column Selection: Choose columns based on application needs, considering factors like particle size, stationary phase, and column dimensions.

- Optimized Conditions: Carefully optimize parameters like mobile phase composition, flow rate, and temperature for optimal separation results.

- Sample Preparation: Ensure proper sample preparation to avoid issues like column fouling and achieve accurate results.

UPLC stands as a potent analytical technique, offering speed, efficiency, and sensitivity in compound separations. This introductory guide provides insights into its principles, benefits, instrumentation, applications, and practical tips for effective utilization.