Has anybody got any experience using the Hi-Plex Na column with a ELSD for oligosaccharide detection? The mobile phase is water for the prescribed column if using RI detector according to Agilent's brochure, but I would like to know what are the operating conditions used if I were to use the ELSD HPLC?
ELSD could be ideal for analyzing many different Compounds. Its response is based on the concentration of the analyte, whether or not it absorbs UV light. It determines sample composition even for unknowns where standards and structural data are not available.
The ELSD’s baseline is quiet and free from drift during gradients. Hence high sensitivity and accuracy. Soi think its possible you can give it a try.
ELSD have good sensitivity than RI. But reproducibility is not so good. Unstable room temperature and gas pressure makes unstable nebulization and makes unstable results. If you want interday reproducibility, you should keep room temperature and use a gas buffer tank.
Hi, thanks for your answers. I am interested to know if anybody has used the ELSD for analyzing oligosaccharides by a ligand exchange Na column and what are the operating conditions (e.g. nebulization temperature, mobile phase flow rate etc.) that I can use as a starting point?
Hi
Some information about separation and detection condition can you find in the following publication: High Performance Liquid Chromatography in Phytochemical Analysis, page 412
http://books.google.de/books?id=8F4LCU-AIhMC&pg=PA410&lpg=PA410&dq=Hiplex+Elsd+varian&source=bl&ots=zGDNDF9ihY&sig=6NYFAPuPqIROdIJaZoeDZ9mz8_g&hl=de&sa=X&ei=Cm29T9CMNcSj8gOthoFJ&ved=0CE0Q6AEwAg#v=onepage&q=Hiplex%20Elsd%20varian&f=false
Moreover, there is an interesting and comprehensive publication of Agilent regarding the combined use of Hi-Plex columns and ELSD detectors, where you can find the columns, conditions used for analysis and detection.
http://www.chem.agilent.com/Library/applications/5990-8801EN%20Hi-Plex%20Compendium.pdf
Check out page 41or at least do a search with your readers (the publication is quite large).
I'm currently using this column for dextrins analysis in wort and beer and also ELSD detection. Column works with water (only water) at 80-85°C and ELSD can manage this mobile phase easily. I set my detector to 2.3psi for nebulizer (nitrogen) and 70°C. be carefoul to not let the column at high temperature without flow and avoid repentine changes in pressure/flow (recomended flow is 0.3 ml/min but I'm working at 0.21 and it works just fine) if you have a precolumn use it
I hope it was helpful to you
regards Mauro
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