Shodex HILIC Column Replacement for ZORBAX Carbohydrate Column (Reproducibility Issues)

Record: 595

Category: Column Selection
Product: NH2P-50 4E 
Sample: Common sugars

Question

A customer is experiencing poor reproducibility with a ZORBAX Carbohydrate column (4.6 × 150 mm, 5 µm) using ACN/water (80:20) with RI detection at 40 °C. The method targets common sugars including fructose, glucose, sucrose, maltose, and lactose (10–1000 µg/mL). Different column lots resulted in shifts in retention time and loss of sensitivity for low-concentration sugars. They would like to maintain the same method conditions and replace only the column. What Shodex column can be recommended?

Answer
For routine HILIC analysis of common saccharides such as fructose, glucose, sucrose, maltose, and lactose, Shodex NH2P-50 4E is typically the recommended starting point. This column provides reliable separation under acetonitrile/water conditions with refractive index detection and is suitable for standard sugar analysis. The application “Yogurt: Sugar added” demonstrates separation of these sugars using NH2P-50 4E under HILIC conditions.

Shodex NH2P-40 3E may be considered when higher resolution or improved sensitivity is required. This column provides enhanced separation performance and improved signal-to-noise, which can be beneficial for low-concentration sugars. The application “Comparison between NH2P-40 3E and NH2P-50 4E” shows improved peak shape and resolution for saccharide analysis using NH2P-40 3E.

Both columns are polymer-based aminopropyl phases designed for HILIC separations and provide more consistent performance compared to silica-based NH₂ columns, which may show variability between lots.

When replacing the column, it is recommended to use the corresponding Shodex NH2P guard column rather than a silica-based NH₂ guard to ensure compatibility and stable performance.

Recommended Applications:

• Yogurt: Sugar Added (NH2P-50 4E)
  Demonstrates separation of fructose, glucose, sucrose, maltose, and lactose under HILIC conditions with RI detection.
• Comparison between NH2P-40 3E and NH2P-50 4E
  Shows improved resolution and peak performance of NH2P-40 3E compared to conventional NH₂-type columns.

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