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.

Updated 01 December 2000.