Chitosan (low molecular weight, deacetylation degree 96.1%, Sigma-Aldrich) was purified according to a previously established method.19 (link) Chitosan (2 g) was dissolved in 200 mL 1% (v/v) acetic acid then filtered (Whatman 541 filter paper). The filtrate was titrated with 1 N NaOH until the pH was approximately 8.5 to precipitate the Chitosan. The precipitate was removed via filtration and resuspended in 500 mL buffer (0.1 M sodium bicarbonate, pH 8.3). Next, 2.5 g sodium dodecyl sulfate (SDS) and 3.72 g ethylenediaminetetraacetic acid (EDTA) were added to the solution and stirred using a magnetic stir bar for 30 minutes. The insoluble Chitosan was filtered, rinsed and dialyzed (Snakeskin) in nanopure water for 24 hours. During dialysis, the water was changed after 10 hours and every hour afterwards. The Chitosan was collected from the dialysis tubing, frozen at −80°C overnight and lyophilized.
Protocol detail
Purification of Chitosan for Research
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Acetic acid
Buffer
Chitosan
Dialysis
Ethylenediaminetetraacetic acid
Filtration
Frozen
Sodium bicarbonate
Sodium dodecyl sulfate
Corresponding Organization :
Other organizations : University of Iowa, Mahidol University, Johns Hopkins Medicine, Johns Hopkins University, Iowa City VA Health Care System
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Variable analysis
independent variables
- Chitosan (low molecular weight, deacetylation degree 96.1%, Sigma-Aldrich)
- Not explicitly mentioned
- Acetic acid concentration (1% v/v)
- NaOH concentration (1 N)
- Sodium bicarbonate buffer (0.1 M, pH 8.3)
- Sodium dodecyl sulfate (SDS, 2.5 g)
- Ethylenediaminetetraacetic acid (EDTA, 3.72 g)
- Dialysis in nanopure water for 24 hours
- Positive control: Not mentioned
- Negative control: Not mentioned
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