Sample Preparation for carbohydrates on the Aminex HPX-87H

Discussions about sample preparation: extraction, cleanup, derivatization, etc.

2 posts Page 1 of 1
My research group is involved in fermentation studies, specifically optimizing and producing bioethanol from feedstocks.

In addition to our Shodex Sugar Column we have recently acquired an Aminex HPX-87H in order to view organic acids, mannitol in addition to glucose, maltose and other standard fermentation sugar products.

From the literature available the conditions for running the column have been set:
The eluant is 5mM sulphuric acid
The flow rate is 0.60 ml/min
The temperature is 50 degrees celsius
The detection is UV 210nm.

Using the above conditions, what would be the best sample preparation for running samples and standards:

1)Firstly we are going to adjust the acidic hydrolysate(as a result of the acidic pretreatment phase) to pH 7, using NaOH
2) We are then going to centrifuge
2) We are then going to filter the sample using a 0.45 µm syringe.


Are there any other recommended internal standards/steps in the sample preparation for optimum HPLC separation and viewing.
Run the sample through an SPE cartridge filled with a cation exchanger in the hydrogen form to get rid of the sodium which otherwise will accumulate on the analytical column. Alternatively, you could use a guard column and change it frequently. I believe BioRad sells a guard column specifically for that purpose.
-- Tom Jupille
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374
2 posts Page 1 of 1

Who is online

In total there is 1 user online :: 0 registered, 0 hidden and 1 guest (based on users active over the past 5 minutes)
Most users ever online was 1117 on Mon Jan 31, 2022 2:50 pm

Users browsing this forum: No registered users and 1 guest

Latest Blog Posts from Separation Science

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.

Liquid Chromatography

Gas Chromatography

Mass Spectrometry