Fig 2. Purification of CS/DS from shortfin mako shark and blue shark fin without skin and cartilage.
(A) Determination of CS/DS in shark fins. Briefly, 5 μg of crude GAGs derived from samples were digested by 50 mU of ChaseACII and 50 mU of ChaseABC, and then resulting unsaturated disaccharides were subjected to reversed-phase ion-pair chromatography with sensitive and specific post-column detection. Peaks: 1, ΔDi-0S; 2, ΔDi-4S; 3, ΔDi-6S; 4, ΔDiUA-2S; 5, ΔDi-diSE; 6, ΔDi-diSB; 7, ΔDi-diSD. (B) Determination hyaluronic acid contents in crude GAGs. Unsaturated disaccharides obtained by digestion with ChaseABC and ChaseACII were analyzed by a graphitized carbon column and post column fluorometric detection. Peaks: 1, ΔDi-0S; 2, ΔDi-HA. (C) Purification of CS/DS from crude GAGs. The crude GAGs (10∼20 mg of dry powder) were purified on DEAE-cellulose column as described under “Materials and Methods”. The dried powders obtained, after desalting and lyophilizing the 6 fraction samples, were weighed. (D) Chromatograms of unsaturated disaccharides of CS/DS derived from shortfin mako shark (Fr. 5) and blue shark (Fr. 3). Peaks: 1, ΔDi-0S; 2, ΔDi-4S; 3, ΔDi-6S; 4, ΔDiUA-2S; 5, ΔDi-diSE; 6, ΔDi-diSB; 7, ΔDi-diSD. Experiments were repeated twice with reproducible results.