Waters ACQUITY UPLC M-Class Columns - Manual (Page 2)

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[ CARE AND USE MANUAL ]
ACQUITY UPLC M-Class Column Care & Use
I. P REPARING ELUENTS
 Do not filter organic solvents. Use MS grade solvents directly
from the bottle.
 A solvent: 100% water with 0.1% formic acid.
 B solvent: 100% acetonitrile or methanol with 0.1% formic acid.
 Seal wash solvent: water, may contain small amount of
organic (no acid).
 Weak needle wash for peptides: 3% acetonitrile with 0.1%
formic acid.
Note:
 Good lab practices: no solvent bottles to go through the
dishwasher!
 Wear gloves when handling solvent lines and hardware.
 If system is contaminated with poor quality solvents,
flush with appropriate solvent.
Many report successful use of solution containing 25% water,
25% acetonitrile, 25% methanol, 25% IPA and 0.1% formic acid.
For PEG contamination, flush system with IPA.
 DO NOT USE strong bases, as they can strip fused silica.
 Where to source solvents:
 In North America: Fisher Optima or Burdick & Jackson.
 In Europe: Biosolve (Netherlands) provide very good solvents.
They offer smaller bottles (0.5 L) as well as “UPLC”
grade solvents.
II. SAMPLE P REPARATION
 Samples for expression analysis must contain tryptic peptides
derived from proteins of interest at suitable concentrations.
 Samples may contain buffers and residual reagents from
approved digestion procedure.
 The sample must not contain other reagents, denaturants,
detergents, lipids, and must be free of particulates.
III. P REPARING AND CONNECTING 75, 100 OR
150 µM I.D. ACQUIT Y UPLC M-CLASS COLUMNS
a) Preparation
Carefully remove your column from bag.
 The gold ferrule on the inlet of the column is placed snugly on the
tubing at the factory. The ferrule is lightly secured to reduce the
risk of it being lost as the column is removed from the packaging.
The ferrule is purposely set high on the tubing but will properly
seat itself in the correct position upon tightening.
 A small piece of Teflon ® tubing is present on the outlet end of the
column for those who want to connect the device to a UV or PDA
detector rather than a mass spectrometer.
Note: The column is attached to a transfer tube with the use of a
zero dead volume union. This is hidden beneath the heat shrink
tubing. Care must be taken NOT to apply excessive force on the
ends of the nano column (e.g., when removing teflon tubing at
column outlet) that could cause this joint to separate.
Nano-Tee with magnetic mount
Analytical column
Tubing to MS source Trapping column
outlet tubing
Tubing to
HTM valve
Inner compartment (swiveled open)