Waters nanoACQUITY UPLC, Quick Start Manual

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4-3
6. Submerge the B1 line in the solvent B reservoir bottle with 0.1% formic
acid/acetonitrile.
7. Place the solvent B reservoir bottle in the solvent tray.
Preparing the sample
The verification test uses the MassPREP Peptide Standard to verify that your
system is operating correctly. The mixture includes nine peptides with a broad
range of polarities and isoelectric points. In addition, there is a void volume
marker. The following table lists the components of the peptide mixture, in
elution order, and their molecular weights.
Tip:
Because Enolase T35, Enolase T37, and Melitin are weaker ionizers in
ESI, they might not be detected.
The peptide mixture contains approximately 1.5
µ
g (~1 nmole) of each peptide.
When reconstituted in 1 mL of solvent (95:5 water/acetonitrile with 0.1%
formic acid), the final sample concentration is ~1 pmole/
µ
L.
MassPREP peptides mixture components:
Peptide
MW,
monoisotopic
(g/mol)
M + 2H M + 3H
Allantoin (V
0
marker) 158.0440
RASG-1 (not trapped) 1000.4938
Angiotensin frag. 1-7 898.4661 450.2401 300.4957
Bradykinin 1059.5613 530.7877 354.1941
Angiotensin II 1045.5345 523.7743 349.5185
Angiotensin I 1295.6775 648.8458 432.8995
Renin substrate 1757.9253 879.9697 586.9821
Enolase T35 1871.9604 936.9872 624.9938
Enolase T37 2827.2806 1414.6473 943.4339
Melitin 2845.7381 1423.8761 949.5864