Troubleshooting and How to Take Care of Your Column and HPLC
Contents
1. Troubleshooting and H ow to Take Care of Your Column and HPLC System HPLC Troubleshooting Pressure too much or too little Leaks pump autosampler detector Reproducibility pump autosampler Column Care Flushing and equilibration D SHIMADZU Solutions for Science Since 1875 Pump Troubleshooting No pressure or fluctuating pressure n Pump may not be completely full of liquid check solvent inlet line n Air in check valve always degas mobile phase n Stuck check valve the pump may have been idle for too long and solvent has dried inside the check valve Poor quality solvent may contain resins that coat the ball __ inside the check valve and that film wont let the ball seat properly D SHIMADZU Solutions for Science Since 1875 Pump Troubleshooting High Pressure n Outlet frit may be blocked with particles from mobile phase or seal material Leaks n Damage to seal and or plunger due to several factors n Misaligned _ plunger n Solvent incompatibility with seal material n Salt crystal buildup from buffers use a rinse kit D SHIMADZU Solutions for Science Since 1875 Pump Troubleshooting Retention Time Reproducibility n For a dual piston pump only one side may be filled with liquid check solvent inlet lines n Temperature change may not be the pump s fault n A 1 shift in temperature can result in a 1 2 shift in retention time n Avoid dra2. fty locations in the lab n Use a column oven when possible SHIMADZU Solutions for Science Since 1875 Autosampler Troubleshooting High Pressure n Particulates from mobile phase sample pump may be trapped in the inlet tubing or valve n Filter mobile phase AND sample when possible Leaks n Fittings may be loose on the valve n Tighten owt te fittings properly and don t exceed the pressure limit of the autosampler SHIMADZU Solutions for Science Since 1875 Autosampler Troubleshooting Area Reproducibility n Always degas rinse phase and use some volume of liquid for rinsing to keep all flow paths in the valves full of liquid n Make sure the needle stroke S deep enough to draw sample from the vial n Check for leaks on the valve fittings and the connection to the column inlet D SHIMADZU Solutions for Science Since 1875 Detector Troubleshooting spiky Baseline n Air bubble in flow cell degas mobile phase n Put some restriction on the cell outlet but not too much Tubing with 0 005 i d is fine Leaks n Cracked flow cell n Don t exceed the pressure limit of the cell n Poor tubing connections n Use the proper fittings and tighten appropriately SHIMADZU Solutions for Science Since 1875 Column Care Follow MER s recommendations for solvent compatibility flow rate and pressure limits Filter samples when possible n Particulates wil
3. l build up on the inlet frit over time Use care when reversing column flow n Connect the outlet to waste NOT inline with the detector to prevent further contamination a Store columns in recommended solvents SHIMADZU Solutions for Science Since 1875 Troubleshooting Summary Throw away bad parts and columns Leaks do not fix themselves If it doesn t pass you must degas D SHIMADZU Solutions for Science Since 1875
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