Chromatography Forum

I have read a number of the postings on the forum and while there are some successes from HILIC applications there appears to be a degree of frustration related to successful operation.

Are there any views on whether HILIC really has a significant future?

I am not sure that this is a valid question.

HILIC is a technique that can solve some problems that are impossible to solve with RP. Example: analysis of sugars. In cases where the problem can besolved by RP and by HILIC, it can have some advantages. Example: sensitivity for cationic analytes with MS detection. Of course, there are also many things that you can't do with HILIC. Example: PAH analysis.

hi interseting that u mentioned HILIC can be used for sugar analysis can u plz elaborate on this....as i am planning flor some sugar analysis method development

I have struggled a bit with the ZIC-HILIC columns, but have not really achieved any useable methods for our hydrophilic drugs. The capacity of the column is low (compared to RP), so the injected amount of substance also have to be low. Not good if you want to detect low level impurities. I have experienced also that the gradients must be slower on HILIC columns (comp to RP).

I have used the RP-CN columns in HILIC mode for sugar analysis, and for that it is very useful since there is no other way to get retention.

Maybe your question should be different:
Will HILIC become more popular than RP?

In this case, I think the answer is no. The mechanism of RP is well known and retention is relative good to predict. HILIC is more difficult to understand.

Nevertheless, like Uwe mentioned, there are defiantly application fields in which HILIC will be much more suited than RP

Rick -

What sugars are you going to be working with?

The standard approach towards sugar analysis in HILIC is the use of an amino column designed for this analysis with a mobile phase of 70% acetonitrile and 30% water. An example is the Carbohydrate Analysis Column from Waters. Do not buy a random amino column, since you will only run into trouble with method stability.

Bryan Evans-

i am working on two method one to resolve and quantification of monosaccrides, and the other on sialic acid estimation...
so got curious with the claim that HILIC can be used for sugar analysis...

Yes HILIC has a future, it will never be as big as RP but as more people are becoming aware of its existance it will continue to grow.
Many of the problems conected to HILIC comes from either trying to separate reactive species or from mixed mode effects due to ionic interactions.
It is also much slower to stabilize after changing eluent composition than RP.

Mattias, I have worked quite a bit with the ZIC-HILIC and never had any problems with capacity can you elaborate?

Bintang> I must admit that I have not given the ZIC-HILIC column so many chances over the years.

I tried to get retention with 5-aminosalicylic acid, and I did. I remember I could not inject close to the usual amount without getting broad and ugly peaks. I am afraid that I cannot remember the details of the separation now. It is quite possible I didn't use the optimal parameter settings.

Are HILIC columns and RP columns generally considered to be equal in tems of mass loadability??

Mass loading per se is not the dominant problem with HILIC. The primary issue is the solubility of the polar compounds that one can separate with the technique in a high acetonitrile content. There may be solutions to this, such as injection in DMSO, but I have not tried them.

Hey Rick -

The monosaccharides can definetly be resolved on a good aminopropyl column. We have a lot of monasaccharide data - if you can tell us which ones you are interested in - we might be able to post some data on here for you.

Sialic acid may also work well on an aminopropyl phase.
As you know, it has functional groups that can be ionized (e.g. COOH group) - so you will need some type of buffer in the mobile phase (as opposed to monosaccharides which do not require a buffer).

Rick -

I forgot, Imtakt does have a sialic acid application using Unison UK-Amino.
This is a low-bleed aminopropyl phase (low noise on ELSD):

http://www.imtakt.com/TecInfo/TI333E.pdf

Sugars can also be analyzed by anion exchange at high pH. Dionex has several CarboPac columns designed for sugar analysis with pulsed amperometric detection.

Bryan Evans-
hi are working on glyco analysis..any data regarding monosaccride analysis will be of great help...
if you have some can u plz mail it abhilashsmenon@gmail.com

hi Mark Tracy
thanks for the suggestion we have just procured dionex sysmtem and its relative columns, although we have not strated any work on them...i was planning to develop a parallel method along with dionex based methos..so was interested in Bryans information...

thank you all