UltiMate 3000 RSLCnano – Standard Applications
48
Table 10: UltiMate 3000 RSLCnano Tandem nano LC kit (P/N 6720.0335) contents - Continued
#
Item Replacement P/N
Cleaving stone 160483
Upgrade kit nano/cap WPS-3000TFC 6824.0030
1/32” 2 pos 6 port nano switching valve 6825.0020
Fittings for nano valve 6720.0080
Polypropylene vials for WPS with glass insert, 250 µL, 25 pcs. 6820.0027
Polypropylene caps for WPS vials, 25 pcs. 6820.0028
Cytochrome C digest, 1.6 nmol, Lyophilized 161089
1/16" Universal Fingertight Fitting, one-piece design, extra long thread, 4 pcs. 6720.0015
2.12.3 Installation Tips
• Follow the General Recommendations for Applications (→ page 6).
• The standard column outlets are 30 cm fused silica capillaries. 20 µm I.D. fused silica
tubing is provided in the kit to extend the column outlets if necessary. Replacing the
attached fused silica by the appropriate length using the nano connector on the column
will give the best result.
• 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
• The WPS-3000FC is normally used for fraction collection. By replacing the divert valve
by a nano valve, the autosampler is fitted for tandem nano LC. Controlling the divert
valve position is performed with the commands Collect and Drain.
2.12.4 Testing the Application
The tandem nano LC setup consists of two pre-concentration nano setups that can be operated
individually; therefore, the system can be tested and evaluated using the conditions in the
table below, as also described in the part of Pre-concentration onto a Nano Column on page
21.
Property Setting
Mobile phase A 100% water + 0.05% TFA
Mobile phase B 20%/80% (v/v) water/ACN + 0.04% TFA
Loading pump A 98%/2% (v/v) water/ACN + 0.05% TFA
Sample Cytochrome C 1 pmol/µL, prepared according to instruction sheet
Injection volume Sample: 1 μL
UV detection 214 nm
Loading time 3 min (may vary with different injection volume/routine)
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