There was an attempt to quantify each components of raffinose family oligosacchrides in comparative study of different legume varieties to identify the variety with less and high quantity of flatus-producing constituents. However, we found most of the sample preparation methods and sequential extraction steps to be very tedious and not inline with the facility in hand.
What is the simplest sample preparation method?
What is the appropriate HPLC column for the separation of raffinose family oligosaccharides(raffinose, stachyose and verbascose) in the extract??
Kindly,
Dear Prof Greg Cote,
Thank you so much for your excellent articles.
Kind regards,
Dear Sir. Concerning your issue about the appropriate HPLC column for the separation of raffinose family oligosaccharides(raffinose, stachyose and verbascose) in the extract. The carbohydrates were separated on a Phenomenex Bondclone 10 μ CHO column (column size 300×3.9 mm). The mobile phase consisted of acetonitrile: water (70:30 v/v) for separation, and the flow rate was fixed at 1 ml/min. The chromatographic data were collected and plotted using Class-VP 6.1 software, and the peak identification of the chromatographs was performed by comparing the retention times with those of standards.The fructose, sucrose, raffinose, and stachyose were all purchased from Sigma Chemicals. I think the following below links and the attached file may help you in your analysis:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC159466/pdf/1051335.pdf
http://www.arccjournals.com/uploads/articles/ArticleFile-LR-3379-7%20(528-532)%20LR-3379.pdf Thanks
Dear Sir
Supelco NH2 analytical column from sigma aldrich-APHERATM NH2 POLYMER
Dear Prof Isam Eldin, and Prof Shekappa, Thank you so much for your contribution. Prof Isam Eldin, so many thanks for detail explanation with instrument control and data handling.
kindly,
Tilahun
Dear Prof Isam Eldin, and Prof Shekappa, Thank you so much for your contribution. Prof Isam Eldin, so many thanks for detail explanation with instrument control and data handling. kindly, Tilahun
Any HILIC column would be suitable. Galacto-oligosaccharides are relatively easy to separate.
The best separations are obtained by anion-exchange chromatography in combination with pulsed electrochemical detection (Dionex system). The detection is many orders of maginitude more sensitive than refractive index, which frequently enables a simple sample treatment. Moreover, a gradient can be used to adjust the separation of lower and higher oligosacharides.
Carbohydrate column is suitable for this purpose. You can refer our publication on raffinose family oligosaccharides of blackgram.
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