8 Run the System
Run the Experiment
2D-LC User Guide 216
Table 26 Recommended conditions in 2D (HPLC) for comprehensive 2D-LC
Parameter Value
2D-LC Valve
MHC with 40 µl sample, Transfer Capillary, ASM Factor No
2
D Column Compartment (MCT)
Column
RRHD Eclipse Plus C18, 3.0x 50 mm, 1.8 µm (959757-302)
Column temperature 40 °C
Stop time As pump/No limit
2
D Pump
Comprehensive
Mobile Phase A Water + 0.2 % formic acid
Mobile Phase B Acetonitrile
Flow Rate 2.5 mL/min
Idle flow not used
Stop time ca. 43 min (will not automatically prolonged, if peaks in 2D are not work off)
Post time 6 min
Sampling Table Start 5 min, Stop at 40 min
2D Gradient: Analysis 0.2 min, Equilibration 0.15 min
Analytical gradient - Shifted Gradient Shift 1D:
25 % B 0.00 min 25 % B 5 min
50 % B 40 min
50 % B 0.2 min 50 % B 5 min
75 % B40 min
2
D Detector (DAD)
Diode-array 254 nm, Bandwidth 4 nm
Reference Wavelength 360 nm
Reference Bandwidth 100 nm
Peak width 80 Hz
Stop time As pump/No limit