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An update for human blood plasma pretreatment for optimized recovery of low‐molecular‐mass peptides prior to CE – MS and SPE – CE – MS
Author(s) -
Pont Laura,
Benavente Fernando,
Barbosa José,
SanzNebot Victoria
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300838
Subject(s) - chromatography , chemistry , protein precipitation , cellulose , size exclusion chromatography , mass spectrometry , peptide , precipitation , filtration (mathematics) , biochemistry , physics , statistics , mathematics , meteorology , enzyme
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 ( A micon® U ltra‐0.5) and 10 K M r cut‐off polyethersulfone filters ( V ivaspin® 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.