Waters ACQUITY UPLC I-Class IVD - Page 24

2Optimizing Performance
24 July 7, 2014, 715003736IVD Rev. C
20 points per second yield higher baseline noise, requiring you to adjust filter
time constants accordingly.
The optimal ACQUITY UPLC flow rate differs from that of a typical HPLC
column. The table below offers operating guidelines for ACQUITY UPLC
columns under both isocratic and gradient conditions. Note that the values
provided are approximations and that optimum performance for your
molecule or separation can occur at a different flow rate.
Follow these general recommendations when performing a UPLC analysis:
•Use high quality solvents, buffers, and additives (MS-grade, prefiltered
with a 2 µm filter).
•Use high quality water (ultrapure, prefiltered with a 2 µm filter).
•Always use solvent filters on tubing lines in solvent bottles.
• Filter buffers with a 0.2-µm filter membrane.
•Keep concentrated stock solutions, to use when preparing working
solutions.
• Do not add fresh buffer to old, which can promote microbial growth.
•Make fresh buffer solutions daily.
•Do not block the degasser vent line. Trim the tubing, if necessary.
•Do not submerge the waste or degasser vent lines in liquid.
See also: For details about how to route the tubing, see the ACQUITY
UPLC I-Class IVD Binary Solvent Manager Overview and Maintenance
Guide (part number 715003739IVD.)
•Flush buffers from the system, using water, immediately after you are
done with them.
•Use 10-20% organic solvent in water as a storage solvent, if you keep the
system idle for extended periods (longer than 24 hours).
Optimal flow rates for molecular weight range:
Column size Molecular weight Flow rate
2.1 × 50 mm <500 600 µL/min
2.1 × 50 mm 1000 300 µL/min
2.1 × 50 mm 1500 150 µL/min
2.1 × 50 mm 2000 100 µL/min