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 Ulset; Bjorn E Christensen; Roger  Andersson; Atte Mikkelson; Päivi  Tuomainen; Ndegwa Henry 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

Anne Rieder, Svein Halvor Knutsen, Ann-Sissel Ulset, Bjorn E Christensen, Roger Andersson, Atte Mikkelson, Päivi Tuomainen, Ndegwa Henry Maina, Simon Ballance (Corresponding Author)

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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.
Copyright © 2015 Elsevier Ltd. All rights reserved.

Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan | Request PDF. Available from: https://www.researchgate.net/publication/273507433_Inter-laboratory_evaluation_of_SEC-post-column_calcofluor_for_determination_of_the_weight-average_molar_mass_of_cereal_b-glucan [accessed Sep 18 2018].

Original language	English
Pages (from-to)	254-264
Journal	Carbohydrate Polymers
Volume	124
DOIs	https://doi.org/10.1016/j.carbpol.2015.02.019
Publication status	Published - Feb 2015
MoE publication type	Not Eligible

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@article{b92f5849e7ce497683621cc686b779c5,

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

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.Copyright {\textcopyright} 2015 Elsevier Ltd. All rights reserved. Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan | Request PDF. Available from: https://www.researchgate.net/publication/273507433_Inter-laboratory_evaluation_of_SEC-post-column_calcofluor_for_determination_of_the_weight-average_molar_mass_of_cereal_b-glucan [accessed Sep 18 2018].",

author = "Anne Rieder and Knutsen, {Svein Halvor} and Ann-Sissel Ulset and Christensen, {Bjorn E} and Roger Andersson and Atte Mikkelson and P{\"a}ivi Tuomainen and Maina, {Ndegwa Henry} and Simon Ballance",

year = "2015",

month = feb,

doi = "10.1016/j.carbpol.2015.02.019",

language = "English",

volume = "124",

pages = "254--264",

journal = "Carbohydrate Polymers",

issn = "0144-8617",

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T1 - Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan

AU - Rieder, Anne

AU - Knutsen, Svein Halvor

AU - Ulset, Ann-Sissel

AU - Christensen, Bjorn E

AU - Andersson, Roger

AU - Mikkelson, Atte

AU - Tuomainen, Päivi

AU - Maina, Ndegwa Henry

AU - Ballance, Simon

PY - 2015/2

Y1 - 2015/2

N2 - 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.Copyright © 2015 Elsevier Ltd. All rights reserved. Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan | Request PDF. Available from: https://www.researchgate.net/publication/273507433_Inter-laboratory_evaluation_of_SEC-post-column_calcofluor_for_determination_of_the_weight-average_molar_mass_of_cereal_b-glucan [accessed Sep 18 2018].

AB - 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.Copyright © 2015 Elsevier Ltd. All rights reserved. Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal β-glucan | Request PDF. Available from: https://www.researchgate.net/publication/273507433_Inter-laboratory_evaluation_of_SEC-post-column_calcofluor_for_determination_of_the_weight-average_molar_mass_of_cereal_b-glucan [accessed Sep 18 2018].

U2 - 10.1016/j.carbpol.2015.02.019

DO - 10.1016/j.carbpol.2015.02.019

M3 - Article

VL - 124

SP - 254

EP - 264

JO - Carbohydrate Polymers

JF - Carbohydrate Polymers

SN - 0144-8617

ER -

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

Abstract

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