The Protein Id Chip, Part No. G4240-62001; Modes Of Operation - Agilent Technologies 1260 Infinity User Manual

1 Introduction to HPLC-Chip Technology
Chip Types

The Protein ID Chip, Part No. G4240-62001

This HPLC-Chip is the standard chip for one-dimensional separations. It is
comprised of a 40 nL enrichment column and a 75 µm x 43mm separation
column which is packed with Zorbax C18-SB 5 µm material. The functionality
of this chip is equivalent to conventional nanospray LC/MS.

Modes of Operation

There are two ways to load the enrichment column. Each has advantages and
disadvantages. It depends on the application which one is preferable. Agilent
recommends to use Forward Flush mode since this mode of operation is less
susceptible to clogging of the enrichment column.
Forward Flush mode
from µ-WPS
from nano pump
Figure 4
Port 6 of the HPLC-Chip is connected to the µ-well-plate sampler. When the
valve is in the loading position, the sample is transferred at a flow of 4 µL/min
onto the enrichment column. Since the loading solvent composition is highly
aqueous, the peptides are retained at the front end of the column. The loading
solvent is directed to waste through port 5. A filter at the back of the
enrichment column retains the packing material.
14
Loading the sample in forward flush mode
to waste
enrichment column
analytical column
HPLC-Chip Cube Interface User's Guide