Established method of chondroitin sulphate extraction from buffalo (Bubalus bubalis) cartilages and its identification by FTIR

Abstract

A study was conducted for extraction of chondroitin sulphate (CS) from buffalo tracheal, nasal and joint cartilages. CS was extracted from cartilages using 0.25% papain digestion, dialyzed, precipitated with 10% TCA and finally lyophilized to dry powder. Dimethylmethylene blue assay was performed to estimate the quantity of CS extracted. Identification of extracted CS was performed with SDS-PAGE and Fourier transforms infrared spectroscopy (FTIR). SDS-PAGE analysis of extracted CS revealed similar electrophoretic pattern to that of standard and the molecular weight ranged from 5 to 20 kDa. FTIR spectra of extracted CS revealed presence of characteristic peaks of –CONH vibration of amide group, coupling of C–O stretching vibration, S=O stretching vibrations and –C–O–S molecules confirms the CS moiety. It can be concluded that extraction method adopted could efficiently be utilized for the extraction of CS from buffalo by-products like tracheal, nasal and joint cartilages.

Fish gills of Thunnus albacares: A novel source of chondroitin sulphate glycosaminoglycans

“…The extracted and purified GAGs produced spectra indicated the presence of methyl groups, carboxyl groups and sulphate groups. Comparable characteristic peaks of -CONH was observed at 1646 cm -1 for standard CS and same peaks of isolated GAG was reported in samples of buffalo tracheal, nasal, and joint cartilages at 1647, 1650, 1650 cm -1 respectively [30] . Characteristic peaks of -CONH vibration of amide group coupling of C-O stretching vibration at 1641 cm -1 were obtained for CS in samples extracted from chicken keel cartilage [31] .…”

Heterologous production of chondroitin

“…The structural composition, molecular weight and yields of CS obtained from extraction vary not only with the extraction method but also with the animal and tissue used. Some of the reported CS yields include: extraction from shark fin resulted in 150.5 mg/g dry cartilage [68] ; from blue shark head wastes 120.8 mg/g dry cartilage was obtained; crocodile hyoid, rib, sternum, trachea cartilages generated 91 – 274 mg/g of dry cartilage [68] ; 75 mg/g of dry weight has been achieved by extraction from ray [68] ; buffalo nasal, joint and tracheal cartilages generated 60 to 62 mg/g of dry cartilage [69] ; using chicken keel, 24.8 mg/g of wet cartilage were obtained [70] ; from different sea cucumbers, fucosylated CS (fCS) yields varied from 63 to 110 mg/g weight [54] ; and from fish by-products, yields from 19 to 137 mg/g dry cartilage have been achieved [71] . In addition to the relatively low yields, the shortage of such materials and the concerns on intraspecies contamination led to studies on artificial synthesis of the chondroitin backbone, either using chemical synthesis, enzymatic, chemo-enzymatic and microbial fermentation strategies.…”

Chondroitin Sulfate and Its Derivatives: A Review of Microbial and Other Production Methods

“…Despite the remarkable strides made towards chondroitin and CS biosynthesis, the yield is still not competitive with the yield from animal-derived CS. On average, when CS is extracted from a buffalo, about 183.28 mg/g of dried cartilage is harvested [74]. This adds up when you consider the weight of the different cartilage types, such as the tracheal, nasal, and joint cartilages of the buffalo.…”