LC-MS
Eliminate Matrix Effects with HybridSPE
Oct 26 2016
HybridSPE®-Phospholipid technology is a sample prep platform designed to selectively remove endogenous protein and phospholipid interferences from biological plasma and serum prior to LC/MS or LC/MS/MS analysis. This technology eliminates ion-suppression and extends column life by eliminating on-column phospholipid buildup. HybridSPE-Phospholipid technology offers exceptional value and simplicity with minimal method development.
Common Issues
Excessive background from endogenous sample matrix components is always of great concern in bioanalysis, alongside the continuing need for decreased analytical run times. One of the major causes of ion-suppression in bioanalysis is the presence of phospholipids during LC/MS analysis. Phospholipids are prevalent in extremely high concentrations in blood-based biological fluids (~1 mg/mL), representing the second largest lipid component in biological matrices after triglycerides.
In addition to ion-suppression, phospholipids may build up on the analytical column. They may elute uncontrollably downstream in an injection run sequence, causing unpredictable ion-suppression effects, poor reproducibility, and decreased data accuracy. If gradient conditions are needed to elute phospholipids, analysis run times are dramatically increased.
Solution
HybridSPE-Phospholipid products are designed for selective removal of both phospholipids and proteins from biological matrices. The procedure involves a few simple steps, analogous to standard protein precipitation. These products are available in both cartridge and 96-well format for compatibility with high-throughput and automation. HybridSPE-Phospholipid technology bridges the gap between simplicity and selectivity, while providing excellent analyte recovery and reproducibility.
Unlike other phospholipid removal products that use a hydrophobic retention mechanism, HybridSPE-Phospholipid technology uses a patented Lewis acid-base retention mechanism, allowing it to separate phospholipids from even very hydrophobic analytes. Competing products (all using hydrophobic interactions for phospholipid removal) retain hydrophobic analytes, such as vitamins, disabling the ability for downstream analysis.
In summary, HybridSPE-Phospholipid products are valuable tools in LC/MS analysis. They help to eliminate both ion-suppression and the need for long gradient run times to elute on-column phospholipid buildup. For additional information, or to request a free sample, please visit the website.
Now Available -- Online HybridSPE for a Hands-Free Workflow
MilliporeSigma is now offering a line of online SPE cartridges, called Supel™ Genie, which include HybridSPE. Following offline protein precipitation, our Supel Genie online HybridSPE cartridges will remove >95% of phospholipids from biological samples, offering a seamless workflow from start to finish performed entirely on the LC instrument.
Online SPE offers all the benefits of offline SPE, along with dramatically increased throughput and reduced labor costs, as the LC instrument automates the entire process. Because the most labor intensive steps of the process are now automated, the risk of human error is eliminated. Visit our website to learn how to streamline your LC/MS workflow.
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