The on-line 2D Nano LC/MS solution consists of a unique instrument configuration:
- The high performance micro autosampler is connected directly to the quaternary pump.
- From the pump the hydraulic flow passes through the micro 6-port/2-position valve of the micro well-plate autosampler and then the sample loop and needle.
- The SCX column is positioned after the injector seat capillary and the SCX column outlet flow is directed to the 6-port valve of the autosampler.
- This valve is connected with a capillary to the micro 10-port/2-position valve in the separate valve box.
- Sample eluting from the SCX column is concentrated on a C18 enrichment column, which is mounted between two ports of this second valve and the eluting flow goes to waste.
- The nanoflow pump is also connected to this second valve and the flow from this pump passes through the analytical column into the Agilent 6500 Series Accurate Mass Q-TOF LC/MS.
- Peptides eluted at the current salt step concentration are trapped on the enrichment column, then the enrichment column is switched into the nanoflow path.
- The nanoflow pump then delivers the elution gradient and the peptides are separated on the reversed-phase nano column.
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