Hollow-fiber flow field-flow fractionation is here applied to untreated, whole human blood serum. Matrix-assisted, laser desorption/ionization time-of-flight-mass spectrometry (MALDI-TOF-MS) of serum fractions shows mass signals in the <30 000 Mr range where low-abundance, serum protein components are known to be present, though a membrane of nominal 30 000 Da cutoff was employed for the fractionation device. Using diluted sera spiked with low amounts (0.06-0.1%, w/w) of an artificial mixture constituted the human adrenocorticotropic hormone fragments 18-39 (Mr = 2465.7) and 7-38 (Mr = 3659.2), and of bovine insulin (Mr = 5734), horse cytochrome c (Mr = 12 384) and chicken lysozyme (Mr = 14 388), a hybrid fractionation/microfiltration mechanism shows to govern the separation of the low-Mr components.
Bibliographical noteFunding Information:
The work was partially supported by the University of Bologna (funds for selected research topics), and by the Italian Ministry of the University and Research (MiUR, PRIN 2006, 2006033944). P.R. and M.-H.M. duly acknowledge the Significant Bilateral Project “Novel Methods for Top-down Analysis of Macromolecular and Nanosized Samples of Biotechnological and Environmental Interest” within the VIII Executive Programme of Scientific and Technological Co-operation between Italy and Korea for the years 2007–2009.
All Science Journal Classification (ASJC) codes
- Analytical Chemistry
- Organic Chemistry