Can I Use a UPLC Column on an HPLC System?

When it comes to Liquid Chromatography (LC) techniques, both High-Performance Liquid Chromatography (HPLC) and Ultra-Performance Liquid Chromatography (UPLC) have become fundamental tools in analytical chemistry. HPLC has been a standard in many laboratories for years, while UPLC is a more recent technology offering faster analyses, higher resolution, and enhanced sensitivity. Given these advantages, many users ask: Can I use a UPLC column on an HPLC system? In this blog post, we’ll explore this question in detail, discussing compatibility, potential benefits, and challenges of using UPLC columns on HPLC systems.

The Difference Between HPLC and UPLC

Before diving into whether a UPLC column can be used on an HPLC system, it’s essential to understand the core differences between High-Performance Liquid Chromatography (HPLC) and Ultra-Performance Liquid Chromatography (UPLC).

HPLC, often referred to as High-Performance LC, typically operates at pressures of up to 6,000 psi and uses columns with particles in the range of 3 to 5 microns. In contrast, UPLC operates at much higher pressures, up to 15,000 psi, and utilizes much smaller particles, typically around 1.7 microns. This enables UPLC, or Ultra-Performance LC, to achieve higher resolution and faster run times compared to HPLC.

Why UPLC Columns Are Not Recommended for HPLC Systems

Now that we understand the fundamental differences between HPLC and UPLC, let’s address why using a UPLC column on a High-Performance Liquid Chromatography system is generally not recommended.

The primary reason for this is pressure. Ultra-Performance Liquid Chromatography is designed to operate at pressures far exceeding what a typical HPLC system can handle. UPLC columns are optimized for higher pressure, enabling faster flow rates and better separation. On the other hand, HPLC systems are limited in their pressure handling capacity, meaning they won’t be able to push the mobile phase through the smaller particles of a UPLC column effectively. As a result, the use of UPLC columns on HPLC systems often leads to underperformance, inefficient separation, or even equipment damage.

Technical Challenges of Using UPLC Columns on HPLC Systems

Using Ultra-Performance LC columns on a High-Performance LC system presents multiple technical challenges, some of which are critical to system performance and durability:

• High Backpressure: UPLC columns generate significantly more backpressure due to their smaller particle sizes. This excess pressure can exceed the capacity of many HPLC systems, potentially causing equipment failure or long-term wear on sensitive components such as seals and pumps.

• Reduced Efficiency: One of the main reasons for using UPLC technology is to achieve faster run times and higher resolution. However, when a UPLC column is used in an HPLC system, these benefits are compromised due to the lower operating pressures. In most cases, the run time increases significantly, and resolution is reduced.

• Longer Analysis Times: Since High-Performance LC systems cannot operate at the pressure required by UPLC columns, the result is often longer analysis times. This negates one of the primary benefits of using Ultra-Performance Liquid Chromatography—speed.

When UPLC Columns Might Be Usable on HPLC Systems

While not ideal, there are some cases where users might still attempt to use Ultra-Performance LC columns on High-Performance LC systems. However, this comes with necessary adjustments and compromises.

• Adjusting Flow Rates: Reducing the flow rate in the HPLC system can sometimes allow for the use of UPLC columns, but this comes with a cost: longer run times. Lowering the flow rate reduces the pressure generated by the system, allowing it to handle the backpressure from the UPLC column. However, this is rarely a sustainable solution for laboratories focused on efficiency.

• Longer Columns: In some cases, using a longer UPLC column can help distribute pressure more evenly when operating on an HPLC system. Yet, this also results in extended analysis times, which contradicts the primary advantage of using Ultra-Performance Liquid Chromatography.

• Columns Calculator: Tools like the Waters Columns Calculator can help users determine whether a particular UPLC column could theoretically work on their HPLC system by adjusting flow rates, column lengths, and particle sizes. However, even with these adjustments, performance is still likely to be compromised.

Why Some Laboratories Consider Using UPLC Columns on HPLC Systems

Given the limitations, why would anyone attempt to use a UPLC column on an HPLC system? In some instances, laboratories may lack access to Ultra-Performance LC equipment but wish to take advantage of the smaller particle sizes of UPLC columns for improved separation. Even if the HPLC system can't fully utilize the potential of a UPLC column, users might still achieve some marginal improvements in separation efficiency and peak resolution for difficult compounds.

However, these gains are often not worth the drawbacks of using mismatched systems.

Risks to Your Equipment

Attempting to use a UPLC column on an HPLC system carries inherent risks. High-Performance Liquid Chromatography systems are not built to handle the high pressures associated with Ultra-Performance Liquid Chromatography columns, and pushing them to do so can lead to:

• System Failure: Overloading the system with excess pressure can cause breakdowns, particularly in components like pumps and seals.

• Reduced Lifespan: Continuous use of high-pressure columns in a low-pressure system can shorten the lifespan of your equipment, requiring costly repairs or replacements.

UPLC Columns on HPLC: A Risky Proposition

In most cases, using Ultra-Performance LC columns on a High-Performance Liquid Chromatography system is not advisable due to the considerable risks and performance compromises. UPLC columns are designed to function optimally in systems that can handle the associated higher pressures and faster flow rates. Forcing them into an HPLC setup diminishes their advantages and may damage your system.

Monad’s Affordable Refurbished Instruments for UPLC Upgrades

For laboratories looking to upgrade from High-Performance LC to Ultra-Performance LC technology without breaking the bank, Monad offers a range of high-quality refurbished instruments. These units provide the speed, resolution, and sensitivity of UPLC at a fraction of the cost of new systems.

Each refurbished instrument from Monad undergoes rigorous testing to ensure it meets the highest standards of quality and reliability. With Monad’s refurbished Ultra-Performance LC instruments, you can enhance your laboratory's capabilities without stretching your budget.

While using a UPLC column on an HPLC system is technically possible, the performance compromises and risks to your equipment make it an unwise practice in most cases. For laboratories looking to enhance their separation efficiency, upgrading to Ultra-Performance Liquid Chromatography is a more practical solution than attempting to use UPLC columns with HPLC systems.

With Monad's refurbished UPLC instruments, you can transition to Ultra-Performance LC technology without the high cost, providing an affordable solution to meet your laboratory’s evolving needs.