Phenomenex Application Note
Philip J. Koerner, J.T. Presley, Tom Cleveland and Jason A. Anspach, Phenomenex Inc., Torrance, California, USA.
For many years, the use of guard columns has been advocated by column manufacturers and other experts to protect and extend column lifetime and performance of analytical columns from potential damage caused by the presence of chemical contaminants and microparticulates in the sample and mobile phase. The introduction and adoption of smaller particle size (< 3 µm) and narrow i.d. columns (≤ 2 mm) for ultra-high performance presents additional challenges. Specifically, these ultra-high performance liquid chromatography (UHPLC) columns are very sensitive even to small increases in dead volume, and the use of traditional guard columns or guard cartridge systems will show a marked decrease in column performance (efficiency and resolution). However, small particle columns are even more susceptible to contamination and blockage from microparticulates that may be present in both the sample and mobile phases as they can occlude the smaller porosity column frits and dramatically shorten column lifetime.
KrudKatcher Ultra in-line filters are specifically designed for use with ultra-high performance columns from 1.0 to 4.6 mm i.d. and are pressure rated to 20000 psi (1375 bar). The filter body houses an integrated 0.5 µm 316 stainless steel depth filter that efficiently removes microparticulates from the flow stream without contributing to system backpressure or dead volume (<0.2 µL).
Figure 1(a)
To illustrate the benefit of using the KrudKatcher Ultra in-line filter for extending column lifetime, a chromatographic separation of a ketone mixture on a Kinetex 2.6 µm PFP 50 × 2.1 mm column was set up on a UHPLC system (Figure 1). The initial injection shows excellent peak shape and was maintained for over 1100 injections [Figure 1(a)]. However, after 1175 injections the chromatographic performance has noticeably degraded resulting in split peaks with some peak broadening and tailing evident [Figure 1(b)]. Simply replacing the KrudKatcher Ultra and re-injecting the sample returns the original chromatographic performance [Figure 1(c)]. The KrudKatcher Ultra in-line filter successfully protected the ultra-high performance Kinetex column from damaging microparticulates in the mobile phase flow path and sample that would have dramatically shortened the column lifetime.
Figure 1(b)
The regular use of KrudKatcher Ultra in-line filters with ultra-high performance UHPLC columns protects and extends the lifetime by protecting them from damaging microparticulates that may be present in the sample and mobile phase. Additionally, the KrudKatcher Ultra does not have a detrimental effect on chromatographic performance (retention time, peak width, resolution, or efficiency).
Figure 1(c)
If you would like more information on KrudKatcher Ultra, please contact your local Phenomenex representative and request technical note TN-1063.
KrudKatcher is a trademark of Phenomenex Inc.
Phenomenex Inc.
411 Madrid Avenue, Torrance, California, 90501, USA
tel. +1 310 212 0555 fax +1 310 328 7768
E-mail: info@phenomenex.com
Website: www.phenomenex.com
AI and GenAI Applications to Help Optimize Purification and Yield of Antibodies From Plasma
October 31st 2024Deriving antibodies from plasma products involves several steps, typically starting from the collection of plasma and ending with the purification of the desired antibodies. These are: plasma collection; plasma pooling; fractionation; antibody purification; concentration and formulation; quality control; and packaging and storage. This process results in a purified antibody product that can be used for therapeutic purposes, diagnostic tests, or research. Each step is critical to ensure the safety, efficacy, and quality of the final product. Applications of AI/GenAI in many of these steps can significantly help in the optimization of purification and yield of the desired antibodies. Some specific use-cases are: selecting and optimizing plasma units for optimized plasma pooling; GenAI solution for enterprise search on internal knowledge portal; analysing and optimizing production batch profitability, inventory, yields; monitoring production batch key performance indicators for outlier identification; monitoring production equipment to predict maintenance events; and reducing quality control laboratory testing turnaround time.
Investigating Synthetic Cathinone Positional Isomers using LC–EAD-MS
November 7th 2024Peng Che fom Vrije Universiteit Amsterdam in the Netherlands discusses the benefits of hyphenating liquid chromatography (LC) with electron activated dissociation mass spectrometry (EAD-MS) to analyze cathinone positional isomers.