What is back pressure in HPLC?
High Back Pressure is defined as an unexpected increase in the pressure readings during normal HPLC operation that approach or exceed the maximum pressure capability of the system. Higher than expected pressure is the most often reported problem in HPLC.
How do you get air out of UPLC?
You shouldnt have to break any connections, just try priming the system for 10 minutes. If you feel you must break a connections, loosen that inlet line right where it goes into the degasser, wait the the bubbles to flush out, retighten, then prime for 1 or 2 minutes.
What causes high back pressure in HPLC?
Causes of Abnormally Elevated Pressure High back pressure in LC instruments is usually caused by foreign material blocking the flow of mobile phase. Although crimped PEEK or stainless steel tubing will occasionally be the culprit, particulates clogging the system are most often the cause.
How do I unclog a column in HPLC?
First, wash the column with varying solvents at a low flow rate. However, if your column is so clogged that it can’t flow at all, flip the column and run it in the opposite direction. If all else fails, then you might need a new column. Like I said earlier, columns do have a finite life.
How do I set up HPLC?
For setting up the HPLC machine:
- Make sure you have all your buffers set up.
- Open the purge valve and purge the system for 5 minutes.
- Add your samples into the autosampler tray.
- Stop the purge.
- Close the purge valve.
- Run the system at a normal flow rate (1 ml/min) with your buffer to equilibrate the column for 10 minutes.
How do I use new column in HPLC?
Connect only the inlet end of the new column to the LC system, and connect spare waste or connecting tubing to the column outlet and allow the column flush effluent to flow to waste. Flush the column with at least 10 column volumes of 100% methanol or 100% acetonitrile—whichever is more convenient.
How do you get air out of HPLC?
Treatment 2: Open the compression fitting at the top of the outlet check valve with a slow pump flow until solvent leaks around the fitting. Tap the side of the check valve with a small wrench until small air bubbles come out with the liquid.
What happens if HPLC runs dry?
If the reservoir runs dry in your HPLC pump you will draw air into the pump, but the pump will not pump air through the system. HPLC pumps are designed to pump liquid, not air. When they fill with air, they lose their prime and stop pumping.