Hello everyone! Throughout my master's thesis and Ph.D. studies, I have encountered difficulties in achieving optimal clean-up of the UHPLC-ESI-MS/MS system.
My research focus lies in the analysis of peptides derived from plasma samples, and despite implementing an extensive pretreatment procedure aimed at eliminating proteins, particularly albumin, the chromatographic system consistently exhibits issues such as increased noise in the chromatogram and elevated column backpressure. Even the use of a pre-column has not provided a complete resolution to these challenges.
Notably, these problems seem to intensify when my colleagues, particularly those working in the metabolomic field, use the instrument following my experiments. Despite employing a thorough cleaning protocol involving a prolonged gradient elution (lasting approximately 1.5 hours) to ensure the removal of all potential samples residues from the system, the results remain the same.
I would like to know if you have come up with the same problems and if you ahve any insights on potential strategies to overcome these persistent issues. Whether it involves refining the pretreatment procedure, exploring alternative stationary phases, optimizing the cleaning protocol of the system, or considering additional precautions for shared instrument use.
Looking forward to hearing your experiences!
Hello Mariano,
It is pretty common to have carry-over analyzing plasma/serum samples. Someone recommended me the use of Trifluoroethanol to wash out all retained peptides. Basically you just use it as a sample and inject 1 or 2ul direct onto the column with a relative short gradient. It does not damage the column in my experience, so maybe you can give it a try in the future.
Good luck!
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