Care and use manual – Waters Glyco-Pak DEAE Columns User Manual
Page 3
[ Care and Use ManUal ]
Glyco-Pak DEAE Columns
3
a. Mode of Action
The Glyco-Pak DEAE column operates in an ion-exchange mode. Retention
times and resolution are very sensitive to small changes in the ionic strength
and or pH of the mobile phase.
b. Typical Chromatograms
Conditions
Sample:
Hydrazine released oligosaccharides from fibrinogen
Mobile Phase:
A: 1 mM sodium phosphate pH 6.0
B: 10 mM sodium phosphate pH 6.0
Gradient:
Isocratic A for 10 min then Linear gradient 100% A
to 100% B over 20 min then hold 100%B for l0 min
Flow Rate:
0.8 ml/min
Detectors:
490 detector 214 nm with baseline subtraction
Figure 2: Hydrazine Released Fibrinogen Oligosacchrides
Conditions
Sample:
Hydrazine released fetuin oligosaccharides
Mobile Phase:
A: water
B: 100 mM NaC1
Gradient:
0-50 % B, Curve 6, 30 min
Flow Rate:
0.6 ml/min
Detection:
490 detector 205 nm with baseline subtraction
Figure 3: Initial Fractionation of Fetuin Oligosaccharides
Conditions
Sample:
Trisialyl-triantennary fetuin oligosaccharides
Eluent:
15 mM Mono-basic sodium phosphate adjusted to
pH 5.4 with sodium hydroxide
Gradient:
Isocratic
Flow Rate:
0.8 ml/min
Detection:
490 at 200 nm
Figure 4: Sub-fractionation of the Trisialylated-triantennary
Fraction of Fetuin