Quantitative analysis of proteins via sulfur determination by HPLC coupled to isotope dilution ICPMS with a hexapole collision cell

Publisher:
ACS Publications
Publication Type:
Journal Article
Citation:
Analytical Chemistry, 2007, 79 (23), pp. 9128 - 9134
Issue Date:
2007-01
Full metadata record
Files in This Item:
Filename Description Size
Thumbnail2010003407OK.pdf125.32 kB
Adobe PDF
Quantitative analysis of proteins is an essential part and also constitutes a major challenge in modern proteomics. Quantification of proteins by inductively coupled plasma mass spectrometry (ICPMS) offers an alternative method for quantitative proteomics. In this study, we developed a method of absolute quantification of proteins via sulfur by size exclusion chromatography (SEC) coupled to ICPMS with a collision cell (ICP-CC-MS) and postcolumn isotope dilution. Bovine serum albumin (BSA), superoxide dismutase (SOD), and metallothionein-II (MT-II) served as model proteins. Enriched 34S, 65Cu, and 67Zn isotopic solutions were continuously mixed with the eluate from the SEC. Oxygen was added as a reactive gas into the collision cell where sulfur reacts with oxygen to form sulfur-oxygen ion, the ratio of 32S16O+/34S16O+ thus representing 32S+/34S+. The absolute quantity of proteins could be calculated by the isotopic dilution equation and the content of sulfur in the proteins. The detection limits for BSA, SOD, and MT-II are 8, 31, and 15 pmol, respectively. The relative standard deviations for the proteins are less than 3%. The ratios of S/Cu and S/Zn in the proteins were also determined. The quantitative method was validated by comparing with gravimetric results.
Please use this identifier to cite or link to this item: