Application of nonporous polyurethane (PU) membranes and porous PU thin films as sample supports for MALDI-MS of wheat proteins

Abstract

Non-porous polyurethane (PU) membranes and porous PU thin films are used as sample supports for MALDI-TOFMS. Mass spectra obtained are compared with those acquired using metal targets and the crushed matrix method. The compounds characterized are wheat proteins which consist of moderately water-soluble gliadins, and of water-insoluble low molecular weight (LMW) and high molecular weight (HMW) glutenins. Mass spectra obtained using the PU supports are in general of good quality, and this method of sample preparation is the most convenient for sample handling. In the case of gliadins and LMW glutenins, the spectra obtained on PU are comparable with those obtained using metal supports. Isolation of the LMW and HMW wheat proteins characterized in this study requires the use of buffers incompatible with MALDI. Spectra of samples containing buffer components on PU supports are of better quality than those obtained using the crushed matrix method. This effect is attributed to stronger protein binding onto the PU supports, which allows for extensive washing and removal of water soluble buffer components. The PU film, when cast onto a MALDI probe, is porous and flat in topology. The differences in surface characteristics between the PU film and the PU membrane result in slight variations in the mass spectra. The extent of surface charging, observed significantly using 50 µm thick PU membranes, decreases with 25 µm membranes and becomes insignificant with PU thin films. An important advantage of using the PU supports is the possibility of preparing samples on the film or membrane in the field and of analysing them at a later time. This is especially important when samples are susceptible to chemical degradation in solution. These proteins are known to degrade while stored in solution. We have thus incorporated the use of PU membrane–film supports into our routine analysis of these proteins.Key words: gliadins, glutenins, MALDI, membrane supports, polymeric supports, time-of-flight analysis.