This document describes a phenol-sulfuric acid assay that is used to detect the presence of sugars such as those found in lipopolysaccharides (LPS) and O-antigens. The assay involves adding phenol and concentrated sulfuric acid to standards of glucose and samples containing LPS, allowing the mixtures to stand for 10 minutes, and then measuring absorbance at 490nm to quantify the amount of sugar present.
This document describes a phenol-sulfuric acid assay that is used to detect the presence of sugars such as those found in lipopolysaccharides (LPS) and O-antigens. The assay involves adding phenol and concentrated sulfuric acid to standards of glucose and samples containing LPS, allowing the mixtures to stand for 10 minutes, and then measuring absorbance at 490nm to quantify the amount of sugar present.
This document describes a phenol-sulfuric acid assay that is used to detect the presence of sugars such as those found in lipopolysaccharides (LPS) and O-antigens. The assay involves adding phenol and concentrated sulfuric acid to standards of glucose and samples containing LPS, allowing the mixtures to stand for 10 minutes, and then measuring absorbance at 490nm to quantify the amount of sugar present.
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.