How do you regenerate a column in HPLC?
Regeneration Procedure for Reversed-Phase Silica Columns
- Disconnect the column and reconnect it to the system, with the flow through the column in the reversed direction.
- Flush out any salts/buffers with HPLC grade water.
- Flush column with 25 mL of isopropanol.
- Flush column with 25 mL of methylene chloride.
How do I protect a column in HPLC?
To protect HPLC columns (analytical and preparative) from contamination, like particulates from the injection, unfiltered mobile phase components or fragments from worn pump seals/ rotor seals from the injector, it is recommended to use a guard column.
How do I clean my IC column?
Inject 1 column volume of pepsin (1 mg/mL in 0.5 M NaCl, 0.1 M acetic acid). Leave overnight at room temperature or for one hour at 37 °C. Rinse with at least 2 column volumes of distilled water (Table 21 for recommended flow) until the UV-baseline and the eluent pH are stable.
How do you clean cyano columns?
- Remove the guard cartridge.
- Flush the column with 100% water for 10 column volumes.
- Inject 200 µL of 50:50 dimethylsulfoxide/water every 5 minutes until 10 column volumes of water has passed through the column.
- Stop injections.
- Flush column with 10 column volumes of 100% water.
How do I clean my HPLC c18 column?
In your case wash the column with 70% water; 15% methanol; 15% acetonitrile. Divert the column eluent to waste not to contaminate your detector(s). Wash the column slowly over to 100% methanol and wash for at least 15 minutes. Wash the column over to 100% acetonitrile and wash for at least 15 minutes.
How should columns be stored?
Columns should be stored at room temperature in their original box, with a copy of the certificate of analysis for future need and to evaluate the column prior to future use.
What is inside a guard column?
It serves to remove the impurities and suspended solids from reaching the analytical column. Typically it has a length of about 2 cm and internal diameter of 4.6 mm. Guard columns are packed with pelicullar particles of around 40 μm size to offer negligible pressure drop.