Abstract: This study aimed to establish the sequential static and static–dynamic supercritical carbon dioxide (SDCO2) fractionation conditions to obtain a higher yield and desired chitosan with lower polydispersity index (PDI) and higher degree of deacetylation (DD). The yield increased with increasing DD of used chitosan and amount of cosolvent. The yield of acetic acid cosolvent was higher than those of malic and citric acid cosolvents. SDCO2, compared to static supercritical carbon dioxide, has higher yield. The yield of extracted chitosan was 5.82–14.70% by SDCO2/acetic acid, which increases with increasing pressure. The DD of fractionated chitosan increased from 66.1% to 70.81–85.33%, while the PDI decreased from 3.97 to 1.69–3.16. The molecular weight changed from 622 kDa to 412–649 kDa, which increased as density of supercritical carbon dioxide increases. Hence, higher DD and lower PDI extracted chitosan can be obtained through controlling the temperature and pressure of SDCO2.