HPLC Analysis of Pullulan on an SRT® SEC-300 Gel Filtration Column
Materials
analytical column
SRT® SEC-300 Gel Filtration Column, 5 μm
L × I.D. 30 cm × 7.8 mm, 5 μm particle size, stainless steelguard cartridge
SRT® SEC-300 Gel Filtration Guard Column, 5 μm
L × I.D. 5 cm × 7.8 mm, 5 μm particle size, stainless steelCONDITIONS
column
SRT SEC-300, 30 cm x 7.8 mm I.D., 5 μm, 300 Å (Z777051)
mobile phase
150 mM phosphate Buffer, pH 7.0
flow rate
1.0 mL/min
pressure
600 psi (41.4 bar)
column temp.
25 °C
detector
RI (30° C)
injection
20 μL
sample
10 mg/mL in mobile phase 0.342 kD, 1.32 kD, 6 kD, 10 kD, 21.7 kD, 48.8 kD 113 kD, 210 kD, 366 kD and 805 kD
Description
Analysis Note
Pullulan is a water-soluble polysaccharide polymer consisting of glucan units. Pullulan is used in the manufacture of edible films that are used in breath fresheners and mints and is also used as a food additive. Another use of pullulan is as calibration standards in gel filtration chromatography.
Gel filtration chromatography (GFC) is used to determine the molecular mass composition of pullulan and many other water-soluble polymers using silica- or polymer-based particles that have been modified with a chemically bonded hydrophilic coating.
In this particular example, a SRT SEC-300 column from Sepax Technologies was used to analyze several pullulan standards. In the accompanying table shows retention volumes for the pullulan standards and the molecular mass of each; this data was then used to construct the calibration curve which allows the calculation of the molecular mass and distribution of an unknown pullulan polymer or other polymers of similar geometrical shape.
Legal Information
SRT is a registered trademark of Sepax Technologies