Is it possible to eliminate only the bulky fructose and glucose (F/G) content from the matrix of interest simultaneously and selectively?
I should get rid of the F/G composition but not the other various mono and oligosaccharides along with similar structure polar metabolites. To find out a convenient polar marker at HILIC condition, removal of F/G is crucial since bulk concentration is suppressing ionization at mass spec. and contaminating the ionization chamber/sampling capillary. Totally, this issue is hampering the marker determination capability of the metabolomic approach.
I look for any advice other than molecularly imprinted polymer (MIP) usage and immunoaffinity-capturing methodology. I have tested lectin affinity resin which unfortunately not selective fructose and MIP showed non-specific binding causing the loss of information.
Any suggestion will be appreciated, thanks in advance...
Check https://www.mdpi.com/2077-0375/10/10/298/pdf
https://www.britannica.com/science/sugar-alcohol
Also check on this paper
https://www.hindawi.com/journals/jchem/2016/6271808/
Maybe an idea is to add Glucose oxidase and fructose oxidase. You will transform selectively F and G into acid form. Then at the end perform a spe separation using polar NH2 column.
Second option: try to separate F/G directly using a polar NH2 column and acetonitrile/water, 80/20 as an eluant.
Regards.
Dr.Elias Bou-Maroun thanks for your suggestion.
Actually, the need for F/G removal is different. Even I convert the glucose ingredient to gluconic acid enzymatically, this ends as a bulky organic acid composition which is also a bit problematic for the HILIC-shotgun metabolomics approach. Since we should apply minimal sample prep to hold metabolite information in the matrix as intact. I do not prefer to perform complicated experimental sample prep.
The purpose is like depletion of the serum abundant proteins to able to discover the whole proteome including rare ones without any matrix suppression. There are lots of options for albumin and/or IgG removal (Cybercon blue resins, albumin removal resins, hexapeptide ligand libraries, proteograph nanoparticle libraries, protein A&G or L ligand attached sorbents, etc..). These can adjust the dynamic range of concentration by removal the abundant proteins by employing different molecular interactions (immunoaffinity, Vroman effect, etc.).
We can implement this elimination strategy in RP mode but not for HILIC due to the bulky F/G are interfering with the assay and masking the polar metabolite detection.
Regarding the NH2 column application. The fractionation could be reasonable. In this case, I may prefer a 2D-LC configuration option which is a seamless way to perform online fractionation.
Thanks for the advice...
Second option: elimination of F/G using SPE normal phase chromatography. Regards.
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