Matrix effect explained by unexpected formation of peptide in acidified plasma

• Aim: Peak distortion and strong signal enhancement was observed when applying a bioanalytical method based on mixed-mode SPE, hydrophilic interaction chromatography and ESI-MS to acidified rabbit plasma samples. Results: High-resolution ESI-MS and N-terminal peptide sequencing revealed a peptide NFQNAL, which was confirmed by H/D exchange ESI-MS. Conclusion: The peptide causing the observed matrix effect was formed by enzymatic degradation of serum albumin at pH 3. Degradation required both acidification and presence of other plasma constituents in addition to albumin to take place. The degree of signal enhancement correlated to the level of NFQNAL in the ion source as measured by MS, with a maximal enhancement factor of 3 at intermediate levels of NFQNAL. The interference was eliminated by changing to another type of hydrophilic interaction chromatography column.

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

acidification; amino terminal sequence; analytic method; animal tissue; Article; blood analysis; blood sampling; chemical analysis; controlled study; electrospray mass spectrometry; enzymatic degradation; hydrophilic interaction chromatography; matrix effect; nonhuman; peptide synthesis; pH; plasma; rabbit; sequence analysis; signal processing; solid phase extraction

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