How can I reduce interfering peaks in RapiFluor-MS blanks when using an ACQUITY Premier Glycan BEH C18 AX column? - WKB231393
ENVIRONMENT
- Mixed-mode chromatography
- Bridged ethylene hybrid (BEH)
- Anion exchange (AX)
- GlycoWorks
- RapiFluor-MS
- ACQUITY Premier Glycan BEH C18 AX
ANSWER
ADDITIONAL INFORMATION
From the article linked above:
“Given its selectivity, mixed mode chromatography was seen to be slightly more prone than HILIC to interferences from sample preparation. Several fluorescently active reagent peaks from RFMS derivatization were identified within the glycan elution window. Most of these peaks were found to result from byproducts of the glycan derivatization reaction and some impurities incorporated from the SPE clean-up step. Each of these interferences could be significantly reduced by adjusting the original RFMS sample preparation protocol [18]. An RFMS byproduct peak exhibited an m/z value of 450.2 Da was successfully removed by applying a repeat SPE clean-up step (Supplemental Figure S5), and another broad interference peak induced by phosphate ions was cleaned away by desalting samples prior to enzymatic deglycosylation (Supplemental Figure S6). These interferences did not interfere with MS detection given than RFMS labeled glycans can be selectively detected with an acquisition window ranging from 700 to 2000 m/z. It is also worth mentioning that the sample loss caused by an extra SPE clean-up step was insignificant. For samples studied in this work, it was found that glycans were obtained with over 90% recovery from 2-passes as compared to 1-pass of SPE. More information about the optimization of SPE procedures can be found in the Supplementary Materials.”
Also see:
MaxPeak Premier | Non-Specific Adsorption – Infographic
ACQUITY Premier C18 AX Charge-Based Glycan Solution Infographic