Quantitative analysis of proteins via sulfur determination by HPLC coupled to isotope dilution ICPMS with a hexapole collision cell.
- Publication Type:
- Journal Article
- Citation:
- Anal Chem, 2007, 79 (23), pp. 9128 - 9134
- Issue Date:
- 2007-12-01
Closed Access
| Filename | Description | Size | |||
|---|---|---|---|---|---|
 | 2010003407OK.pdf | 125.32 kB |
Copyright Clearance Process
- Recently Added
- In Progress
- Closed Access
This item is closed access and not available.
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: