Combination of gel electrophoresis and ICP-mass spectrometry-novel strategies for phosphoprotein measurement.
The potential for developing improved procedures for phosphate measurement through combinations of gel electrophoresis and quadrupole-based ICP mass spectrometry utilising (47)PO(+) is investigated. Laser ablation of gels offers a rapid and direct quantitation route, but is subject to high blanks due to P impurities in gels and associated reagents; nevertheless optimisation of laser sampling afforded improved method sensitivity (limit of detection 0.09 microg g(-1)). Implementation of whole gel elution (WGE) with FI-ICP-MS (conventional solution nebulisation) following gel electrophoresis permitted quantitation at the sub microg l(-1) level, and microcolumn processing (activated alumina) was effective at rejecting phosphate contamination. The potential for S-induced molecular ion interference at mass 47 was demonstrated.
Centre for Analytical Sciences, Department of Chemistry, University of Sheffield, Dainton Building, Sheffield, S3 7HF, UK.
Victoria L
VL
Elliott
Cameron W
CW
McLeod
Peter S
PS
Marshall
D002138:Calibration; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007834:Lasers; D013058:Mass Spectrometry / Q000379:methods*; D008956:Models, Chemical; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification
D010750:Phosphoproteins