Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan

Anne Rieder; Svein Halvor Knutsen; Ann-Sissel T Ulset; Bjørn E Christensen; Roger Andersson; Atte Mikkelson; Päivi Tuomainen; Ndegwa Maina; Simon Ballance

doi:10.1016/j.carbpol.2015.02.019

Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan

Carbohydr Polym. 2015 Jun 25:124:254-64. doi: 10.1016/j.carbpol.2015.02.019. Epub 2015 Feb 21.

Authors

Anne Rieder¹, Svein Halvor Knutsen², Ann-Sissel T Ulset³, Bjørn E Christensen⁴, Roger Andersson⁵, Atte Mikkelson⁶, Päivi Tuomainen⁷, Ndegwa Maina⁸, Simon Ballance⁹

Affiliations

¹ Nofima, Norwegian Institute for Food, Fisheries and Aquaculture Research, PB 210, N-1431 Ås, Norway. Electronic address: anne.rieder@nofima.no.
² Nofima, Norwegian Institute for Food, Fisheries and Aquaculture Research, PB 210, N-1431 Ås, Norway. Electronic address: svein.knutsen@nofima.no.
³ NOBIPOL, Department of Biotechnology, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway. Electronic address: ann-sissel.t.ulset@ntnu.no.
⁴ NOBIPOL, Department of Biotechnology, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway. Electronic address: bjorn.e.christensen@ntnu.no.
⁵ Department of Food Science, Swedish University of Agricultural Sciences, PO Box 7051, SE-750 07 Uppsala, Sweden. Electronic address: roger.andersson@slu.se.
⁶ VTT Technical Research Center of Finland, PO Box 1000, FI-02044 VTT Espoo, Finland. Electronic address: atte.mikkelson@vtt.fi.
⁷ Department of Food and Environmental Sciences, University of Helsinki, PO Box 66, A3039 Helsinki, Finland. Electronic address: paivi.tuomainen@helsinki.fi.
⁸ Department of Food and Environmental Sciences, University of Helsinki, PO Box 66, A3039 Helsinki, Finland. Electronic address: henry.maina@helsinki.fi.
⁹ Nofima, Norwegian Institute for Food, Fisheries and Aquaculture Research, PB 210, N-1431 Ås, Norway. Electronic address: simon.ballance@nofima.no.

PMID: 25839819
DOI: 10.1016/j.carbpol.2015.02.019

Abstract

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 10-500×10(3)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.

Keywords: Calcofluor; Cereal β-glucan; Chromatography; Light scattering; Molar mass.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography, Gel
Flour / analysis
Hordeum / chemistry
Hordeum / metabolism
Light
Refractometry
Scattering, Radiation
beta-Glucans / chemistry*
beta-Glucans / isolation & purification

Substances

beta-Glucans