Phenol Sulphuric Acid Carbohydrate Assay

Reference:

Anal. Chem. 28:(1956) 350-356.

Objective:

To detect the presence of sugars. (Widely used method to monitor column eluates for elution of sugar containing compounds such as LPS, 0-antigen, etc.)

Materials:

• Standards:glucose 1 mg/ml stock solution
• use 10, 20, 40, 60, 80 µl and make up each sample with dH₂O to a final volume of 100 µl.
• Blank 100 µl water
• Samples:50 µl (of 5 mg/ml LPS solution) and make up to 100 µl.

Method:

1. Add 50 ul of 80% Phenol solution (80% Phenol by weight)
2. Vortex. Add 2.0 ml concentrated Sulphuric Acid in a stream.
3. Stand 10 min. at room temperature.
4. Read absorbance an 490nm.