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alot of colum use in hplc
Many different columns could be used in HPLC. It depends on the characteristic of molecules upon which you wish to separate. For example, if you wish to separate molecules based on their polarity, you should use a normal phase column (non-polar elutes first) or reverse phase column (polar elutes first).
There are many types of columns. Here are a few columns and their uses:
- size exclusion: depends on molecular size ( not weight!)
- ion exchange: separates positively and negatively charged molecules
- bioaffinity: depends on the strength of affinity towards a specific biological molecule
Different type of column available for HPLC.
First we know about sample nature like,
Matrix
Structure
Molecular weight
polarity
PH
solubility
Then only we go to column selection.
in HPLC packed columns or liquid bonded phased columns are used.
There are different types of HPLC coulms, mostley differing in stationary phase polarity. Most commonley used is C18. You always check the polarity of analyte of interest and then seasrch for the right coulmn with those preferencies.
for HPLC, the colum is made of stainless steel to support the heigh pressure, the colum is filled by stationnary phase
Silica based Luna C18 (250 mm ×4.6 mm ×5μm) column is one of the column used in HPLC.
