Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan
Publication details
Journal : Carbohydrate Polymers , vol. 124 , p. 254–264 , 2015
Publisher : Elsevier
International Standard Numbers
:
Printed
:
0144-8617
Electronic
:
1879-1344
Publication type : Academic article
Links
:
DOI
:
doi.org/10.1016/j.carbpol.2015...
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Kjetil Aune
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Summary
Even though size exclusion chromatography (SEC) with post column addition of calcofluor (SEC-calcofluor) has been used for the determination of cereal β-glucan molar mass in foods for many years, there is a lack of systematic evaluation of the method. To address this issue a set of suitable β-glucan standards were generated by preparative SEC and their molar mass characteristics were determined by analytical multi-detection SEC (refractive index (RI), light scattering). Each standard was then analysed by SEC-calcofluor at three different labs. As a direct comparison, the analyses were repeated with a RI detector. For SEC-calcofluor accurate measurements of weight average molar mass (Mw) can be made for β-glucan populations within 10–500 × 103 g/mol. Above this molar mass threshold there is an increasing tendency for underestimation of Mw. Precipitation of some β-glucan-calcofluor complexes may have delayed their transport into the detector.