Preparing for Operation 47
3
Priming and equilibrating procedures are described in Section 3.2.1 and Section 3.2.2.
Eluent Guidelines
When preparing a 1525µ or 1525EF Pump for operation, follow these guidelines:
• Use methanol to prime a new pump, or one that has been stored dry for an extended
period.
• Use an intermediate solvent when changing between eluents that are not miscible.
• Use only HPLC-grade eluents to obtain accurate, reproducible chromatography.
• To prevent air from being drawn into the pump, keep the eluent level in the reservoir at
least 4 inches (10 cm) higher than the inlet manifold. Do not place eluent reservoirs
on top of the pump.
• Use only filtered, degassed eluents.
• After use with a buffer, purge the pump with HPLC-grade water.
• When possible, use eluents of similar viscosity when you run a gradient.
• Know the properties of your eluents; for example, cyclohexane solidifies at 3.5 K psi.
• Consider column chemistry. If the column can be damaged by an intermediate or new
eluent, remove the column from the system and substitute a union. Refer to the care
and use guide for your column.
• Dedicate an eluent supply tube and filter assembly for each eluent. If this is not
possible, purge the tube and filter of any remaining eluent before you use a new
eluent.
• To prevent cross contamination of eluents, dedicate a set of glassware for preparing,
storing and supplying each eluent. After a container is used for an eluent, do not use
it for any other eluent unless it has been thoroughly cleaned.
Table 3-1 Pump Preparation Recommendations
Status Prime Equilibrate
Initiating operation
Changing eluent
Adding fresh eluent
System has been idle for some time
Caution:
To avoid chemical hazards, observe safe laboratory practices when handling
eluents. Refer to the Material Safety Data Sheets for the eluents in use. For more
information on eluent properties, see Appendix C,
Solvent Considerations.
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