Isolation, Purification and Structural Identification of Heparinoids from Fish Swim Bladder

doi:10.7506/spkx1002-6630-20180728-351

Abstract

Abstract: In the present study, heparin-like compounds were isolated from fish swim bladder by enzymatic hydrolysis, and they were purified and fractionated into four fractions (F1–F4) by anion exchange resin adsorption followed by sodium chloride gradient elution and alcohol precipitation. Each fraction was identified by cellulose acetate electrophoresis. Furthermore, the structural characteristics of F4 were determined, and the homogeneity and molecular mass were analyzed by UV spectroscopy and high performance gel chromatography. Its monosaccharide composition was analyzed by high performance liquid chromatography (HPLC) after pre-column derivatization, functional group structure by Fourier transform infrared (FTIR) spectroscopy, disaccharide composition by enzymatic cleavage and tandem mass spectrometry (MS/MS), and primary structure by nuclear magnetic resonance. The results showed that the yield of F4, with a purity of up to (85.79 ± 0.63)% was (2.21 ± 0.03) mg/g of dried fish swim bladder. Its molecular mass was about 84 000 u. Monosaccharide analysis revealed that F4 was composed of glucuronic acid (GlcA) and N-acetylgalactosamine (GalNAc) with a small amount of aldoluronic acid and galactose. Its electrophoretic shift was similar to that of chondroitin sulfate and it had the characteristic absorption peaks of carboxyl, acetyl amino and sulfate groups (axial stretching). Finally, F4 was identified as chondroitin sulfate-A composed of repeated disaccharide units of [→4GlcUA β1→3GalNAc(4S)β1→].

Key words: fish swim bladder, heparinoid, isolation and purification, structural identification

CLC Number:

TS254.1

ZHOU Siyi, ZHONG Saiyi, SU Weiming, DU Zhenxing, CHEN Jianping, HONG Pengzhi, ZHANG Chaohua. Isolation, Purification and Structural Identification of Heparinoids from Fish Swim Bladder[J]. FOOD SCIENCE, 2019, 40(15): 84-91.

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Isolation, Purification and Structural Identification of Heparinoids from Fish Swim Bladder

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