What is HILIC column?
Our broad variety of hydrophilic interaction liquid chromatography (HILIC) columns provides strong retention of compounds that are unretained under conventional reversed phase conditions, without the requirement for ion-pair additives in the mobile phase. These columns are ideal for high sensitivity LC-MS applications.
What does HILIC stand for?
Hydrophilic interaction liquid chromatography (HILIC) is an alternative high-performance liquid chromatography (HPLC) mode for separating polar compounds.
How do you condition a HILIC column?
In order to condition your column prior to use, it must be flushed with the mobile phase that will be used during analysis. For isocratic HILIC methods, at least 50 column volumes should be used and, for gradient HILIC methods, at least 10 blank injections running the full-time program should be performed.
How do you clean HILIC columns?
A suitable wash solution is 60% organic solvent in water (unless otherwise specificed in product manuals). HILIC columns should be stored under HILIC conditions. A mixture of organic solvent and water in a 90:10 ratio is recommended for storage.
Is HILIC normal phase?
HILIC is a variation of normal phase chromatography. The major differences between HILIC and NP are the composite of the mobile phase and the mechanism of separation. Common NP chromatography uses 100 % organic mobile phases while HILIC uses organic mobile phases that are water miscible.
What is HILIC mode?
What is HILIC mode? Hydrophilic Interaction Chromatography. It is a separation mode utilizing the interaction difference of hydrophilic properties between the compounds. The elution order in the Hilic mode is in reverse order of reversed phase mode. That is to say from least polar to most polar.
Is HILIC reverse phase?
HILIC can be described as a variation of reversed phase chromatography performed using a polar stationary phase. The mobile phase employed is highly organic in nature (> 60-70% solvent, typically acetonitrile) containing a small percentage of aqueous solvent/buffer or other polar solvent.
How does HILIC chromatography work?
Hydrophilic interaction chromatography (HILIC) is a liquid chromatography (LC) technique that uses a polar stationary phase (for example, silica or a polar bonded phase) in conjunction with a mobile phase containing an appreciable quantity of water (usually at least 2.5% by volume) combined with a higher proportion of …
How do you increase peak shape in HILIC?
Insufficient buffer concentration can result in increased secondary interactions that cause peak tailing. Increasing the buffer concentration can enhance hydrogen bonding between analyte and stationary phase, masking other secondary interactions and improving peak shape.
Does acetonitrile dissolve silica?
You can use of of the following solvents to remove anything left in the column: Ethyl acetate or acetonitrile, those are really the most efficient solvents used in organic synthesis and silica gel chromatography. 30% methanol in DCM could dissolve the silica gel TLC. Akash M Sarkar please see the posts above.
How do you clean column chromatography?
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. Depending on the type of column you are using you can store the column in 100% organic depending on the manufacturer’s recommendations.
What is the difference between HILIC and normal phase?