Which column is best for HPLC?
A Quick Look at Reversed Phase HPLC
- C18 HPLC columns are the most preferred as they offer an excellent range of hydrophobic separation power along with high surface area coverage.
- C8 HPLC columns are used when less retention compared to a C18 is needed.
What does C18 mean in HPLC column?
C18 simply means that the molecules contain 18 carbon atoms, so the other atoms in the molecule can vary, leading to significantly different substances. Knowing the characteristics of the compounds that will be run through the C18 columns can help you choose which one will work best for you.
What is hypersil column?
Hypersil APS columns are a versatile aminopropyl group stationary phase which can be used in 3 modes: ion chromatography (weak anion exchanger), reversed phase and normal phase chromatography. They also offer an alternative selectivity to silica when used in normal phase.
How do I restore a column in HPLC?
How do I restore my HPLC column?
- Routinely monitor the column’s performance.
- Switch only between miscible mobile phases.
- Avoid precipitating salts in the column.
- Use filtered and degassed mobile phases.
- Do not allow the column to dry out.
- For prolonged storage, use a mobile phase that inhibits bacterial and mold growth.
How many types of HPLC columns are there?
High performance liquid chromatography (HPLC) is a type of liquid chromatography that uses a liquid moblie phase. The same basic principals from gas chromatography are applied to liquid chromatography. There are three basic types of liquid chromatographic columns: liquid-liquid, liquid-solid, and ion-exchange.
What is a reverse column?
A reverse phase column, or reversed-phase HPLC columns, are chromatography columns that contain a non-polar stationary phase. A sample is placed into a reverse phase column and then solvent is added to flush the sample through the stationary phase.
Is C18 column polar or nonpolar?
A C18 column is an example of a “reverse phase” column. Reverse phase columns are often used with more polar solvents such as water, methanol or acetonitrile. The stationary phase is a nonpolar hydrocarbon, whereas the mobile phase is a polar liquid.
How do you regenerate a column?
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.
What is a column volume?
Column volume is the volume capacity of your column and bed volume is the volume of resine inside. Mostly they are the same.
How long do HPLC columns last?
Normal phase column have slightly shorter life spans, but should accept about 1000 injections. When subjected to extreme conditions, most columns will be stable for only a few hundred injections or less.
