UHPLC Column Protection Dramatically Extends Performance and Lifetime - - Chromatography Online
UHPLC Column Protection Dramatically Extends Performance and Lifetime

The Application Notebook


UHPLC columns can significantly improve chromatographic separations, but they also present unique challenges. Once the UHPLC system components are optimized, perhaps the greatest concern is protecting the column from the damaging effects of microparticulates and sample contaminants.

An ultra-high performance column protection system, specifically designed for UHPLC systems using sub-2 m and core-shell particle columns, can be used to extend column lifetime (saving both money and time through less frequent column replacement), while minimizing system troubleshooting and downtime.

Results and Discussion

It is well known that UHPLC systems and columns require higher levels of care and attention than traditional HPLC in order to reap their full ultra-high chromatographic performance benefits. Once system components are optimized, chromatographic cleanliness is vital to maintain UHPLC performance, and column protection is compulsory. Unprotected columns may suffer from reduced performance and lifetime, and may lead to an increased need for system troubleshooting and/or downtime.

UHPLC columns packed with sub-2 m particles tend to clog much more rapidly than traditional HPLC columns packed with larger 3 m and 5 m particles. This may be due to the fact that, not only is the interstitial space between the particles much smaller, but columns packed with sub-2 m particles also use frits with a much smaller porosity compared to conventional HPLC columns. With a "tighter", more restricted flow path, any undissolved matter or particulates from the sample, the mobile phase or the system (such as micro-particulates shedding from piston seals and injection valve rotors), will quickly and irreversibly foul the UHPLC column.

Figure 1: Scanning electron microscopy (SEM) of contaminated and non-contaminated column inlet frit.
An easy way to extend the performance and lifetime of UHPLC columns (either sub-2 m fully-porous or core-shell media) is to prevent any contaminants from reaching the column by using the SecurityGuard ULTRA guard cartridge system (Figure 1).

Figure 2: Accelerated column lifetime test.
Presented in Figure 2 is an accelerated lifetime test using an endogenous biological matrix injected onto a coreshell column (Kinetex 2.6 m C18 50 4.6 mm column). With the unprotected column (grey dots), sequential injections of the matrix lead to a steady and irreversible increase in back pressure. Without SecurityGuard ULTRA column protection, the increase in back pressure becomes exponential. This increase in back pressure will eventually lead to degraded chromatography, including band broadening and possibly peak splitting. As a result, method sensitivity, quantitation and peak identification may also be adversely affected.

However, column lifetime is greatly extended by using the SecurityGuard ULTRA (red boxes). In this case, sequential injections of the matrix will still lead to an increase in pressure, but this is due to the particulates being captured in the SecurityGuard ULTRA itself, rather than in the UHPLC/HPLC column. Thus, by simply replacing the SecurityGuard ULTRA cartridge at regular intervals, back pressure returns to starting levels and effective column lifetime can be greatly extended.

Phenomenex Inc.
411 Madrid Ave., Torrance, California, USA
tel: +1 310 212 055 fax: +1 310 212 7768
Website: http://www.phenomenex.com/


blog comments powered by Disqus
LCGC E-mail Newsletters
Global E-newsletters subscribe here:



Column Watch: Ron Majors, established authority on new column technologies, keeps readers up-to-date with new sample preparation trends in all branches of chromatography and reviews developments. LATEST: When Bad Things Happen to Good Food: Applications of HPLC to Detect Food Adulteration

Perspectives in Modern HPLC: Michael W. Dong is a senior scientist in Small Molecule Drug Discovery at Genentech in South San Francisco, California. He is responsible for new technologies, automation, and supporting late-stage research projects in small molecule analytical chemistry and QC of small molecule pharmaceutical sciences. LATEST: HPLC for Characterization and Quality Control of Therapeutic Monoclonal Antibodies

MS — The Practical Art: Kate Yu brings her expertise in the field of mass spectrometry and hyphenated techniques to the pages of LCGC. In this column she examines the mass spectrometric side of coupled liquid and gas-phase systems. Troubleshooting-style articles provide readers with invaluable advice for getting the most from their mass spectrometers. LATEST: Radical Mass Spectrometry as a New Frontier for Bioanalysis

LC Troubleshooting: LC Troubleshooting sets about making HPLC methods easier to master. By covering the basics of liquid chromatography separations and instrumentation, John Dolan is able to highlight common problems and provide remedies for them. LATEST: How Much Can I Inject? Part I: Injecting in Mobile Phase

More LCGC Columnists>>

LCGC North America Editorial Advisory Board>>

LCGC Europe Editorial Advisory Board>>

LCGC Editorial Team Contacts>>

Source: The Application Notebook,
Click here