The EASY-nLC combines simplicity and high-throughput capacity in a nano-flow UHPLC with a pressure rating of 1200 bar, here is a link to Thermo's website if you wish to learn more: EASY-nLC™ 1200 System

• EASY-nLC 1 (LC-030755 / UW tag 2033411), Installed 10/16/2018
  MasterKey: d01ZhUfrREe9XOaH
• EASY-nLC 2 (LC-030757 / UW tag 2033412), Installed 10/19/2018
  MasterKey: M0yI2sgti5yDw6vI
• EASY-nLC 3 (LC-030758 / UW tag 2033413), Installed 10/19/2018
  MasterKey: zBh7kbe82Wl8IQEU
• EASY-nLC 4 (LC-031111 / UW tag 2033416), Installed 12/15/2020
  MasterKey: KZSF7nwD2Q30KCl+
• EASY-nLC 5 (LC-031112 / UW tag 2033417), Installed 12/15/2020
  MasterKey: REEkEDaHVr6P2WiP
• EASY-nLC 6 (LC-031113 / UW tag 2033418), Installed 12/15/2020
  MasterKey: Eh0p+iwjW5reU6Iz
• EASY-nLC 7 (LC-031114 / UW tag 2033419), Installed 12/15/2020
  MasterKey: Z8CaplMxt0ImZkfm

Virtual Display

• Open "Instrument Configuration", select the EASYnLC and Configure
• Test Connection and select the plate installed and click ok and done.
  
  
  
  
  
• Open Putty by double clicking on "putty.exe" located on the desktop
• Under Sessions/Saved Sessions select "EASYnLC" and click "Load" and then "Open"
• login as: hplc
• Password: hplchplc
• use the up arrow to reload the last command or type: x11vnc -q -display :0
  after hitting enter you should see this:
  
  
  
  
  
• If you don't see that type this alternate command (EASY-nLC 7 (LC-031114)): x11vnc -env FD_XDM=1 -auth guess
  
  
• Open "TightVNC Viewer" located on the desktop
• In the "TightVNC Server" field type: localhost::5901
• then hit "connect" to open the virtual display of the EASY-nLC touchscreen
  
  
  
  
  Initial setup of Putty and the virtual display (e.g. on a new PC):
  Guide how to initially setup the virtual display (pdf)
  
  

Switch between Trapping and No-trapping mode

• Looking at the EASY-nLC home page you can see if the system is in trapping or no-trapping mode. It either displays the trap before the analytical column or not.
• To switch the trapping mode, click on the Maintenance/Devices/EAY-nLC [HPLC]/Properties
• To select no-trap mode check the box next to “One column setup”
• To select trapping mode, uncheck the next to “One column setup”
• Hit “Apply” and accept the popup confirmation window
  
  
  
  
  
• Now make sure the plumbing is setup correctly
• In trap mode put a trap column in front of the analytical column
• In no-trap mode replace the trap column with an empty fused silica



• Now make sure the sample loading in the method is setup correctly
• For no-trap mode we recommend loading at lower flowrates e.g. 0.3 µl/min, or set the max pressure instead e.g. 150-200 bar
• the loading volume should be: 2 x sample volume + 2µl
• But always use a minimum of 6 µl (1-2 µl of sample volume)
• e.g. if you load 4 µl of sample: 2 x 4 µl + 2 µl = 10 µl
  
  
• For trap mode you can load at higher flowrates e.g. 2.5 µl, or set the max pressure instead e.g. 150-200 bar
• the loading volume should be: 2 x sample volume + 2µl
• But always use a minimum of 6 µl (1-2 µl of sample volume)
• e.g. if you load 4 µl of sample: 2 x 4 µl + 2 µl = 10 µl

Changing the plate configuration

• Close Xcalibur, methods, qual browser etc.
• You can leave the virtual display and Tune window open
• Open instrument configuration (should be pinned to the start menu)
• Select the EASYnLC and hit configure
• Click on “Test configuration” wait for the “connection OK” turn green
• Under Autosampler Configuration, go to the plate installed and select the plate format you want to use
• Note our systems are only calibrated for 6x8 vials and 96 well plate
• You can click on Edit to see the layout or change the row/number layout if you wish
• Hit “OK” and then “Done” to close the instrument configuration window
  
  
  
  
  
• Now you have to change the plate on the HPLC itself:
• On the EASYnLC home screen click on the plate
• Click on Goto Calibration
• Alternatively, you can go to Maintenance/Devices/Cooled Autosampler/Tools
• Select the plate you want to use from the pull-down menu
• Hit “Use” and acknowledge the pop up
  
  
  
  
  
• Now re-open Xcalibur and you should be able to select the sample positions that correspond to your plate
• 6x8: A1-A8, B1-B8, … F1-F8
• 8x12: A1-A12, B1-B12, …. H1-H12
• Note the vial position for the 6 vials on the right side is 1-V1 through 1-V6

LogViewer

• Log Viewer usage(pdf)

• EASY_nLC Log Viewer 2.18 (zip)

HPLC setup

• LC-MS setup procedure EASYnLC(pdf)
• Simple peptide standards for LC-MS (xlsx)

Instrument Method setup guidelines

• Note: the injection volume in the Xcalibur sequence overwrites the Sample pickup volume in the method
• You need to make sure the at the Sample loading Volume in the method is sufficient to load the entire sample onto the column
• Always set the Sample loading Volume = 2 x injection volume + 2 µl
• But always use a minimum of 6 µl (1-2 µl of sample volume)
• e.g. if you load 4 µl of sample: 2 x 4 µl + 2 µl = 10 µl
• The basic method provided by UWPR will have a loading volume of 20 µl to allow for 1-8 µl sample injection volumes (set in Xcalibur sequence list)

• Note the buffer composition used on the EASY-nLC:
• Buffer A: 0.1 % Formic Acid in Water (Fisher Water with 0.1% Formic Acid (v/v), Optima™ LC/MS Grade, LS118)
• Buffer B: 0.1 % Formic Acid, 80% Acetonitrile in Water (Fisher Water with 0.1% Formic Acid (v/v), Optima™ LC/MS Grade, LS122500)
• Formula 100% ACN to 80% conversion: % B [80%ACN] = 1.25 * % B [100%ACN]

Solvent filters

Upgrading Software/Firmware on the EASYnLC 1200

Troubleshooting

Convert bar to psi