An update for human blood plasma pretreatment for optimized recovery of low-molecular-mass peptides prior to CE-MS and SPE-CE-MS

作者：Pont Laura; Benavente Fernando; Barbosa Jose; Sanz Nebot Victoria^*

来源：Journal of Separation Science, 2013, 36(24): 3896-3902.

DOI：10.1002/jssc.201300838

• CZE
• Biological fluids
• Clean-up
• Preconcentration
• Pretreatment

Protein precipitation and centrifugal filtration are well-established methods for concentrating and purifying peptides with a low relative molecular mass (M-r) from human blood plasma before proteomic and peptidomic studies using high-performance separation techniques, but there is little information on peptide recoveries. Here, we evaluate acetonitrile precipitation followed by a range of centrifugal filtration conditions for the analysis of low M-r peptides in human blood plasma before CE-MS and SPE coupled online to CE-MS. Three opioid peptides were used as model compounds, that is, dynorphin A 1-7, endomorphin 1, and methionine enkephalin and 3, 10, and 30 K M-r cut-off cellulose acetate filters (Amicon (R) Ultra-0.5) and 10 K M-r cut-off polyethersulfone filters (Vivaspin (R) 500) were studied. Unexpectedly, recoveries and repeatability were only optimum after passivating the 10 K M-r cut-off cellulose acetate filters with PEG to avoid peptide adsorption on the inner walls of the plastic sample reservoir.

• 出版日期2013-12