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Extraction and biochemical characterization of sulphated glycosaminoglycans from chicken keel cartilage
 
Humaira Majeed Khan1, Muhammad Ashraf2, Abu Saeed Hashmi3, Mansur-ud-Din Ahmad4 and Aftab Ahmad Anjum5
1Lahore College for Women University, Jail Road, Lahore; 2Department of Pharmacology & Toxicology; 3Institute of Biochemistry & Biotechnology; 4Department of Epidemiology & Public Health and 5Department of Microbiology, University of Veterinary and Animal Sciences, Lahore-54000, Pakistan
*Corresponding author: mansuruddin@uvas.edu.pk

Abstract   

The present study was conducted to explore the potential and cheaper source of major and abundantly found sulphated Glycosaminoglycans in chicken keel cartilage. Chicken is comparatively readily accessible to all the communities of Pakistan and its cartilages are the rich source of sulphated GAGs. The GAGs were extracted from prewashed and ground keel cartilages (n=3) of chicken using 3 M MgCl2, dialyzed, digested with papain, precipitated with three volumes of ethanol, and finally lyophilized to dry powder. The dry products were used for proximate analysis (carbohydrates 65.49±0.10, crude protein 12.82±0.26, ash 11.12±.56, moisture 9.88±0.32 and fat 0.69±0.14%). Dimethylmethylene blue binding (DMMB) assay was performed to determine the quantity of total GAGs in each group of product and protein contents were estimated by Bradford method. Identification of extracted samples of GAGs was performed with FTIR spectrometer using KBr disc and purity of the samples was determined by SDS-PAGE. Quantity of total GAGs in extracted samples was 70.77±2.27% and estimated amount of protein was 4.64±0.29%. FTIR spectra of standard and samples of CS showed identical and characteristic peaks in finger print region. Finger print region revealed the presence of C-O-S, S=O, -COO, -C-C, R-SO2–R, -CONH2 and R-SO2-NH2 molecules. SDS-PAGE analysis revealed the presence of 77.8 and 50.5 kDa proteins in all extracted samples of GAGs. It can be concluded that chicken keel cartilage is the potential and cheap source of GAGs. Analysis by SDS-PAGE revealed that most of the non-collagen protein can be removed by three volumes of solvent extraction and FTIR is an advance technique for identification of GAGs in mid infrared region (400-4000 cm-1).

Key words: Bradford’s method, Dimethylemethylene Blue Binding (DMMB) Assay, Fourier Transform Infrared Spectroscopy (FTIR), Glycosaminoglycans (GAGs), SDS-PAGE

 
   

ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)



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