RP-HPLC columns can be calibrated in terms of glucose units for day-to-day
comparisons; the order of elution is the same as for NP-HPLC with the lowest
molecular weight gluco-oligomers coming first as verified by MALDI-TOF MS of
three of the standard peaks. Some modifications have also a major effect on
elution times on RP-HPLC, as exemplified by the elution of
Manα1,6(Manα1,3)Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAc-PA
before and after bovine α-fucosidase digestion, which results in a
reduction in elution time from 9.5 to 7.2 glucose units on an RP-amide column
(11.5 to 7.2 g.u. in the case of a standard C18 column); the example glycan
shown is from Trichuris suis.(17)