General Guidelines For A Microscale System; Required Solvents - Waters ACQUITY UPLC M-Class System Manual

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Note:
If the changes in lab temperature are too large, consider relocating the system to a more
temperature-stable area of the lab, or contact the lab facilities manager to improve lab
temperature stability.

2.2 General guidelines for a microscale system

Separations performed on the ACQUITY UPLC M-Class system use smaller ID columns when
compared to an analytical UPLC system. Solvent and sample consumption are significantly
reduced.
The microscale chromatograph requires optimal performance from the µSM-FL because sample
dispersion is more evident when using smaller columns.
Recommendation:
0.5 seconds wide. For good quantitation, a sampling rate of 10 to 15 points across a peak is
desirable.

2.2.1 Required solvents

Important:
Use only high-quality solvents.
See also:
Appendix D: Solvent considerations
Requirements:
• Use high-quality solvents, buffers, and additives (LC/MS-grade, pre-filtered).
• Use high-quality water (LC/MS-grade).
Follow these general recommendations when performing a microscale analysis:
• Always use solvent filters on tubing lines in solvent bottles.
• Filter salt-containing buffers with a 0.2-µm filter membrane.
• Keep concentrated stock solutions to use when you prepare working solutions.
• Make fresh buffer solutions daily.
• To avoid promoting microbial growth, avoid adding fresh buffer to old buffer.
• Keep the seal wash line primed.
• Prime solvent lines during system start-up.
• Keep all solvent lines primed.
• Do not block the degasser vent line. Trim the tubing, if necessary.
Note:
Eluent lines A1 and B1 do not pass through the degassers; however the washes are
degassed. When working at flow rates less than 1µL/min, passing the eluent through the
degasser may cause depletion of any volatile mobile phase modifier. For most applications, a
Regarding fast, microscale analyses, a peak of interest can be less than
November 26, 2019, 715003588 Revision C
Page 25

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