Demo 1 (N-Glycan Standards)

To test the Peak Detection Wizards validity, an equimolar mixture of N-glycan standards (Figure 1) has been analysed. In order to simulate high-throughput conditions low mass concentration and a short time of mass spectrum acquisition has been chosen.
These glycan standards have been dissolved each in MeOH/H2O 1/1(v/v) at the concentration of 0.5 pmol/µl and submitted to nanoESI Q-Tof MS analysis in the positive (Demo 1(+) mode MS), and in the negative ion mode (Demo 1(-) mode MS) simulating a high-throughput procedure.

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The positive ion mode of the mixture of neutral N-glycan standards.

At three scans, which correspond to 3 seconds time acquisition, oligosaccharide standards have been detected as a group of singly and doubly charged ions at different signal abundances.

Peak Detection Wizard parameters: Peak Detection: Demo 1 (+) mode MS.

To demonstrate MS/MS interpretation approach the precursor ions at m/z 1485 (NGA2F) and 1663 (NA2) have been fragmented and submitted for de novo N-glycan automated MS/MS interpretation.

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The same mixture was analysed at the negative ion mode and the spectrum corresponding to 3 seconds time acquisition was processed by SysBioWare

At three scans, which correspond to 3 seconds time acquisition, oligosaccharide standards have been detected as a group of singly and doubly charged ions predominantly showing chloride and phosphoric acid negative adducts.

To demonstrate MS/MS interpretation approach the precursor ions at m/z 1497 (NGA2F) and 1675 (NA2) have been fragmented and submitted for de novo N-glycan automated MS/MS interpretation.

Figure 1