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.
authors.affiliation: Nofima, Norwegian Institute for Food, Fisheries and Aquaculture Research, PB 210, N-1431 Ås, Norway. Electronic address: anne.rieder@nofima.no.
authors.forename: Anne
authors.initials: A
authors.lastname: Rieder
authors.affiliation: Nofima, Norwegian Institute for Food, Fisheries and Aquaculture Research, PB 210, N-1431 Ås, Norway. Electronic address: svein.knutsen@nofima.no.
authors.forename: Svein Halvor
authors.initials: SH
authors.lastname: Knutsen
authors.affiliation: NOBIPOL, Department of Biotechnology, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway. Electronic address: ann-sissel.t.ulset@ntnu.no.
authors.forename: Ann-Sissel T
authors.initials: AS
authors.lastname: Ulset
authors.affiliation: NOBIPOL, Department of Biotechnology, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway. Electronic address: bjorn.e.christensen@ntnu.no.
authors.forename: Bjørn E
authors.initials: BE
authors.lastname: Christensen
authors.affiliation: Department of Food Science, Swedish University of Agricultural Sciences, PO Box 7051, SE-750 07 Uppsala, Sweden. Electronic address: roger.andersson@slu.se.
authors.forename: Roger
authors.initials: R
authors.lastname: Andersson
authors.affiliation: VTT Technical Research Center of Finland, PO Box 1000, FI-02044 VTT Espoo, Finland. Electronic address: atte.mikkelson@vtt.fi.
authors.forename: Atte
authors.initials: A
authors.lastname: Mikkelson
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki, PO Box 66, A3039 Helsinki, Finland. Electronic address: paivi.tuomainen@helsinki.fi.
authors.forename: Päivi
authors.initials: P
authors.lastname: Tuomainen
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki, PO Box 66, A3039 Helsinki, Finland. Electronic address: henry.maina@helsinki.fi.
authors.forename: Ndegwa
authors.initials: N
authors.lastname: Maina
authors.affiliation: Nofima, Norwegian Institute for Food, Fisheries and Aquaculture Research, PB 210, N-1431 Ås, Norway. Electronic address: simon.ballance@nofima.no.
authors.forename: Simon
authors.initials: S
authors.lastname: Ballance
mesh_terms: D002850:Chromatography, Gel; D005433:Flour / Q000032:analysis; D001467:Hordeum / Q000737:chemistry / Q000378:metabolism; D008027:Light; D012031:Refractometry; D012542:Scattering, Radiation; D047071:beta-Glucans / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D047071:beta-Glucans
keywords: Calcofluor; Cereal β-glucan; Chromatography; Light scattering; Molar mass