The weight-average molecular weight (Mw), number-average molecular weight (Mn = ∑NM/∑N, N: molecular weight of each component; M: the number of moles of each component), polydispersity (Mw/Mn) and chain conformation of LBPs were estimated by high-performance size exclusion chromatography using an instrument equipped with a multi-angle laser light-scattering system with a refractive index detector (HPSEC-MALLS-RI, Wyatt Dawn Heleos-II, Wyatt Technology, Santa Barbara, CA, USA) according to the method described by Arakawa [50 ], with some modifications. Isocratic elution with 0.20M NaCl solution at a flow rate of 0.5 mL/min was performed on a Shodex SB-804 HQ and SB-806 HQ (Showa Denko KK, Tokyo, Japan) at 25 °C. Sodium chloride solution (0.20M NaCl) was used as the mobile phase at a flow rate of 0.5 mL/min. The injection volume was 50 µL and running time was 100 min. ASTRA software (version 7.1.2, Wyatt Technology) was used to obtain the data.
Hpsec malls ri
Manufactured by Wyatt Technology
Sourced in United States
The HPSEC-MALLS-RI is a laboratory instrument that combines high-performance size exclusion chromatography (HPSEC), multi-angle laser light scattering (MALLS), and refractive index (RI) detection. This instrument is used for the characterization of the size, molecular weight, and concentration of macromolecules in solution.
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2 protocols using hpsec malls ri
1
Molecular Weight and Conformation Analysis of LBPs
2
Molecular Weight Analysis of Starch Pastes
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The molecular weight of LS and LS-GMS pastes were measured using the method reported by Zeng et al. (2015) . An aliquot of the filtered sample (300 μL) was subjected to HPSEC-MALLS-RI (Wyatt Technology, Santa Barbara, CA, USA) after passing through a 0.22-μm nylon filter. The HPSEC system consisted of P8514-806 columns (Showa Denko, Tokyo, Japan), MALLS (632.8 nm; DAWN DSP; Wyatt Technology), and an RI detector (Optilab DSP; Wyatt Technology). The mobile phase consisted of dimethyl sulfoxide with 50 mmol/L LiBr; the flow velocity was 0.35 mL/min, and the dn/dc value was set at 0.074. Further, the molecular weight (M w ), mean square radius (R z ), and polydispersity (Mw/Mn) of the starch pastes were determined by the designated software.
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