UltiMate 3000 RSLCnano – Standard Applications
31
2.8.3 Installation Tips
• Follow the General Recommendations for Applications (→ page 6).
• The design of the nano trap column provides the easiest connections, but must only be
used in forward flush operation. Please check the indicated flow direction when
installing a nano trap column.
• If the loss of hydrophilic peptides is observed, the concentration of acetonitrile in the
loading solvent can be decreased down to 99/1 water/ACN + 0.05% TFA
2.8.4 Testing the Application
The pre-concentration setup can be tested using the following conditions:
Property Setting
Mobile phase A 100% water + 0.05% TFA
Mobile phase B 20%/80% (v/v)water/ACN + 0.04% TFA
Loading solvent 98%/2% (v/v) water/ACN + 0.05% TFA
Sample Cytochrome C digest 8 pmol, prepared according to instruction sheet
Injection volume 1 μL (partial loop fill of a 20 µL loop)
UV detection 214 nm
Loading time 3 min (may vary with different injection volume/routine)
Gradient 4% to 55% B in 30 min
90% B for 5 min
25min equilibration
WPS temperature
5°C (WPS-3000(B)T only)
Loading flow 20 μL/min
Flow rate 4 μL/min (capillary flow selector)
To Next Page
Loading...
