Antioxidant properties of chitosan from crab shells
Introduction
Chitin, found in the shell of crustaceans, the cuticles of insects, and the cell walls of fungi, is the second abundant biopolymer in the nature (Knorr, 1984). Structurally, chitin is a straight-chain polymer composed of β-1,4-N-acetylglucosamine and classified into α-, β- and γ-chitin (Cabib, 1981, Cabib et al., 1988). Chitosan derived by partial N-deacetylation of chitin is also a straight-chain polymer of glucosamine and N-acetylglucosamine (Muzzarelli, Rochetti, Stanic, & Weckx, 1997). α-Chitin, the most abundant in nature, has a structure of antiparallel chains and is found in the crab, shrimp and lobster whereas β-chitin found in squid has intrasheet hydrogen bonding by parallel chains (Jang et al., 2004, Minke and Blackwell, 1978). However, γ-chitin found in the cell walls of fungi has a mixture of parallel and antiparallel chains, which is a combination of α-chitin and β-chitin (Jang et al., 2004).
Because chitin and chitosan possesses many beneficially biological properties such as antimicrobial activity (Kobayashi et al., 1990, Tokoro et al., 1989), biocompatibility, biodegradability, hemostatic activity and woundhealing property, much attention has been paid to its biomedical applications (Farkas, 1990, Fleet and Phaff, 1981). Due to these unique properties, chitosan and its derivatives have been proposed for applications in biomedical, food, agriculture, biotechnology and pharmaceutical fields (Felse and Panda, 1999, Kumar, 2000, Shahidi et al., 1999).
Antioxidant properties of chitosan derivatives have been studied (Lin and Chou, 2004, Xie et al., 2001, Xing et al., 2005). Furthermore, antioxidant properties of fungal chitosan from shiitake stipes have also been studied (Yen, Tseng, Li, & Mau, 2007). However, antioxidant properties of chitosan derived from crab shells are not available. Accordingly, the objective of this study was to assess the antioxidant properties of chitosan prepared from crab chitin by N-deacetylation using a concentrated sodium hydroxide solution. Antioxidant properties assayed were antioxidant activity by the conjugated diene method, reducing power, scavenging abilities on radicals and chelating ability on ferrous ions.
Section snippets
Chemicals
Methanol was purchased from Mallinckrodt Baker, Inc. (Phillipsburg, NJ). Ascorbic acid, butylated hydroxyanisole (BHA), citric acid, crude crab chitin, crab chitosan, 5,5-dimethyl pyrroline-N-oxide (DMPO), 1,1-diphenyl-2-picrylhydrazyl (DPPH), ethylenediaminetetraacetic acid (EDTA), ferrozine, linoleic acid, potassium ferricyanide, potassium permanganate and α-tocopherol were purchased from Sigma Chemical Co. (St. Louis, MO). Ferrous chloride and hydrogen peroxide were obtained from Merck Co.
Antioxidant activity
Using the conjugated diene method, all crab chitosans exhibited showed moderate to high antioxidant activities of 58.3–70.2% at 1 mg/mL and high antioxidant activities of 79.9–85.2% at 10 mg/mL (Fig. 1). All chitosans showed consistent antioxidant activity with increased concentration. In addition, the antioxidant activities of chitosans C60, C90 and C120 correlated with their N-deacetylation times. However, at 0.1 mg/mL, the antioxidant activities were 89.2%, 88.9% and 32.0% for BHA, α-tocopherol
Conclusion
The results showed all crab chitosans were good in antioxidant properties, especially antioxidant activity, scavenging ability on hydroxyl radicals and chelating ability on ferrous ions. In addition, the prolonged N-deacetylation resulted in chitosan with more effective antioxidant properties. All crab chitosans exhibited comparable antioxidant properties. On the basis of the results obtained, crab chitosan with presumed antioxidant properties may be used as a source of antioxidants, as a
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