Samples were injected onto a Propac-PA1 column (4 × 150 mm, Thermo) affixed with a guard column (4 × 50 mm, Thermo) on an Ultimate 3000 UHPLC system (1.0 mL/min, gradient from 0 to 1.0 mM NaCl in 50 mM NaOH) with fluorescence detection (λex = 348 nm, λem = 440 nm)45 (link). Retention times for the disaccharides were confirmed using anthranilamide labelled disaccharide standard mixes (Galen). Absolute quantification of disaccharides was based on calibration curves constructed with known quantities of disaccharide.
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Disaccharide Quantification by UHPLC
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Corresponding Organization : Scripps Institution of Oceanography
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Variable analysis
independent variables
- Concentration of NaCl (from 0 to 1.0 mM)
- Retention times of disaccharides
- Absolute quantification of disaccharides
- Column type (Propac-PA1 column, 4 × 150 mm, with a guard column, 4 × 50 mm)
- Flow rate (1.0 mL/min)
- Buffer composition (50 mM NaOH)
- Fluorescence detection (λ_ex = 348 nm, λ_em = 440 nm)
- Anthranilamide labelled disaccharide standard mixes (Galen)
- Not explicitly mentioned
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