3 Installation Completion and System Verification
Step 1. Condition the Agilent LC instrument
50 Agilent 6400 Series Triple Quad LC/MS System Installation Guide
Step 1. Condition the Agilent LC instrument
Before you begin, check that you have:
• 18 M, HPLC grade deionized water (p/n 8500-2236)
• high-purity HPLC grade isopropanol
• premixed LC flushing solvent (50:25:15:10
isopropanol/acetonitrile/cyclohexane/dichloromethane)
(p/n G1969-85026)
• high-purity HPLC grade acetonitrile (p/n G2453-85050)
• high-purity HPLC grade methanol (p/n 8500-1867)
• high purity formic acid (p/n G2453-85060)
• for 6470/6495: RRHD Zorbax Eclipse Plus C18 trapping column
(p/n 959757-302)
1 Flush the LC system.
This flushing procedure is very effective at reducing background
contaminants from the LC systems when they are interfaced with a mass
spectrometer. The flushing procedure must be done for each solvent
channel to ensure contaminants are removed from separate degasser and
pumping channels.
2 Remove any column(s), install a restriction capillary, and bypass the flow
cell for the UV detector, unless it is rated for high-pressure (400 bar)
operation. Route the outlet capillary to a waste container.
3 If required by your degasser model, connect A1/A2 degasser channels in
series and B1/B2 degasser channels in series. Use the c-tubes included with
the degasser.
Don't try to re-use solvents used for the flushing procedure.
4 Use a hand-held controller or the Data Acquisition program to set up the
pump module to deliver deionized water for 15 minutes at 3 mL/minute to
remove any salt buffers. Do 5 injections with deionized water to clean the
injector path and sample loop.
5 Set up the pump module to deliver HPLC grade isopropanol for 5 minutes at
3 mL/minute. Do 5 injections with isopropanol to clean the injector path
and sample loop.
6 Set up the pump module to deliver the premixed LC flushing solvent
(50:25:15:10 isopropanol/acetonitrile/cyclohexane/dichloromethane) for 15