Quantitation of binding, recovery and desalting efficiency of peptides and proteins in solid phase extraction micropipette tips.
Micropipette-tip solid phase extraction (SPE) systems are common in proteomic analyses for desalting and concentrating samples for mass spectrometry, removing interferences, and increasing sensitivity. These systems are inexpensive, disposable, and highly efficient. Here, we show micropipette-tip solid phase extraction is a direct sample preparation method for (14)C-accelerator mass spectrometry (AMS), removing salts or reagent from labeled macromolecules. We compared loading, recovery and desalting efficiency in commercially available SPE micro-tips using (14)C-labeled peptides and proteins, AMS, and alpha spectrometry ion energy loss quantitation. The polypropylene in the tips was nearly (14)C-free and simultaneously provided low-background carrier for AMS. The silica material did not interfere with the analysis. Alpha spectrometry provided an absolute measurement of desalting efficiency.
Palmblad
Magnus
M
Center for Accelerator Mass Spectrometry, Lawrence Livermore National Laboratory, Livermore, CA 94551-0808, USA. palmblad1@llnl.gov
Vogel
John S
JS
D004864:Equipment and Supplies; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
D010455:Peptides; D011506:Proteins