Date: 10-SEPTEMBER-2026 Last Updated: 10-SEPTEMBER-2026
The analysis of glucose and galactose can be challenging because these highly polar sugars lack strong UV-absorbing chromophores and differ only in the stereochemistry of a single carbon atom. This close structural similarity makes chromatographic separation difficult using conventional HPLC methods. HILIC chromatography coupled with evaporative light scattering detection (ELSD) provides an effective approach for the retention, separation, and detection of these closely related C-4 epimers.
Peaks: 1. Glucose 2. Galactose
Column: Cogent Diamond Hydride™, 4 μm, 100 Å
Catalog No.: 70000-15D
Dimensions: 4.6 x 150 mm
Mobile Phase:
92%
Acetonitrile / 8% DI water / 0.1%
Formic acid
(v/v)
Flow Rate:
1.0 mL/minute
Detection:
ELSD with a Gain 12, Temp 65C, Nitrogen: 3.5 lb
Injection Volume:
1 uL
Sample Preparation:
A Glucose and Galactose sugar mixture was prepared by combining 100 µL of glucose (4.12 mg/mL), 100 µL of galactose (4.08 mg/mL), and 800 µL of 50:50 diluent containing FA, yielding a final volume of 1.0 mL.
Additional Info: The galactose peak exhibits partial splitting due to the presence of α- and β-anomeric forms in solution. These anomers exist in equilibrium but can interact differently with the stationary phase under HILIC conditions, resulting in partial chromatographic resolution. This behavior is common in carbohydrate analysis and does not indicate sample impurity.
Note: Galactose is one of the two monosaccharides that make up lactose (milk sugar). During digestion, lactose is broken down into glucose and galactose, which are then absorbed and metabolized separately.
