Chromatography Forum

Will this method work?
A: acetonitrile
B: 20 mm ammonium acetate pH10, 5% acetonitrile
Column: 50x2 3um Luna NH2 amino
1 ml/min, 0-1min, 90%A; 2-5min, 90-10%A; 5-8min, 10%A; 8-10 min, 90%A

saying if it will work for something we not know about is quite a request

the pH is quite high but still within the limits set by phenomenex
columns do not work as a HILIC mode over 50% of the buffer phase
but if you get a good chromatography result, separation and reproducibility of your compounds should it matter?
simply know that you are not doing only HILIC there

After using this method and obtaining good peaks, I start to realise that it is not a Hilic method. It is an anion exchange method. Have anyone had any experience with apHera High pH amino polymeric Column and Shodex amino polymetric column? I want to avoid the unstability of silica column under basic condition. These are the polymetric version of amino columns.

I found both of those columns to be really good for HILIC of sugars using an ELSD. Baseline noise was about 10 lower than
when using a silica amino column. I actually preferred the Tosoh amide column and had good luck with the new Waters
acquity amide phase the handful of times that I used it before it died.

Thanks. I will look into those amide columns.

Another possibility, use a polymer-silica hybrid amino column so you can get the best of both, high efficiency and reproducibility.

Bonna-Agela Durashell NH2 or Chromenta EP-NH2 both have 3um and 5um particles. Unison UK-Amino columns are also aqueous durable in 3um particle size.

depending on what you do
SAX WAX should be the way to go since they have an amine group
recently we use Sepax,
they have good columns and they have a 5 or 3u columns
ph range is of 2-12
we found that for such applications to us a non porous column was better

but there are other alternatives and comparable columns as well out there

I found both of those columns to be really good for HILIC of sugars using an ELSD. Baseline noise was about 10 lower than
when using a silica amino column. I actually preferred the Tosoh amide column and had good luck with the new Waters
acquity amide phase the handful of times that I used it before it died.

Sepscientologist - just curious what you meant by your statement that you had good luck with the new Waters amide before it died. What do you mean it died? How many samples did you inject? What were the samples? What were the analytical conditions? My colleague and I have been trying to validate and robustness test a BEH amide 2.1 x 100 mm method (from a Waters application note) for the analysis of sugars and short chain oligomers. We haven't injected any real world fermentation samples, yet, as the column overpressurizes after ~100 standard injections. We cannot figure it out. There are several papers in the literature that describe the use of the BEH amide, under similar conditions, for the analysis of long chain oligosaccharides. I just cannot understand what is going on. Can a column life only be 100 injections? What happens to the phase of the column to cause it to "plug" and refuse to be rescued?

Any thoughts you have on the matter are greatly appreciated. We are pretty frustrated.