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Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering SEC-MALS
Published on: June 20, 2019
Strategy to improve the characterization of chitosan by size exclusion chromatography coupled with multi angle laser
Yu Kang1, Xiaoxue Wu2, Xiangling Ji1
1State Key Laboratory of Polymer Physics and Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China; University of Science and Technology of China, Hefei 230026, China.
Abstract:
Here we present SEC-MALLS study for chitosan samples with weight average molar mass Mw between 33 and 427 kg/mol on columns packed with 8 μm porous particles. A low injection concentration on the order of 0.1-0.2 mg/mL must be used to avoid overloading of the SEC columns, due to the extended coil conformation of cationic chitosan in the dilute acid buffer as compared to neutral polymers. Additionally, SEC must be performed at an eluent flow rate no more than 0.5 mL/min for high molar mass chitosan samples. At flow rates of 1.0 and 1.5 mL/min, the elution of the largest chitosan species is delayed, which leads to a distortion of the molar mass distribution towards lower molar mass region. Such an abnormal behavior is due to a chromatographic mode transition from SEC to slalom chromatography, originated from the coil-stretch transition of chitosan chains in elongational flow through the packed columns.
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