-
- Posts: 74
- Joined: Thu May 09, 2019 7:36 pm
I am analyzing sugars using a HILIC column and notice significant tailing as I increase column temperature from 25C,30C, 35C, 40C.
Discussions about HPLC, CE, TLC, SFC, and other "liquid phase" separation techniques.
Andy Alpert wrote:
I defer to LMH regarding the instrumental issues. As to the chromatography:
1) What column are you using for HILIC?
2) What's your mobile phase(s)?
3) Exactly which sugars are tailing? Are any sugars not tailing? If so, then which ones?
lmh wrote:
Andy Alpert will probably give you the right answer when he notices that this is a Hilic-related thread. But I'd ask how good your column oven and solvent pre-heater are? Hilic is very dependent on things like pH, and pH is very dependent on temperature, and temperature on the outside of a column isn't necessarily the same as that on the inside of the column, if the solvent isn't perfectly pre-heated. If the solvent bottles are sitting at room temperature, and the column oven is set to 25, then everything's more or less the same temperature and there will be only very small temperature gradients, no matter how bad the column oven and preheater. As you raise the temperature, the oven and preheater have more work to do to make sure everything stays coordinated, and temperature differentials will get bigger, as will any problem they cause.
Separation Science offers free learning from the experts covering methods, applications, webinars, eSeminars, videos, tutorials for users of liquid chromatography, gas chromatography, mass spectrometry, sample preparation and related analytical techniques.
Subscribe to our eNewsletter with daily, weekly or monthly updates: Food & Beverage, Environmental, (Bio)Pharmaceutical, Bioclinical, Liquid Chromatography, Gas Chromatography and Mass Spectrometry.