ISOLATION, PURIFICATION, AND CHARACTERIZATION OF PORCINE SKIN COLLAGEN: ANALYSIS OF THE GLYCINE, PROLINE, AND HYDROXYPROLINE COMPONENTS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
Objective: The aim of this study is to produce collagen through the extraction and isolation of porcine skin.
Methods: Collagen from porcine skin (Sus scrofa domesticus) was isolated, purified, and characterized. Major amino acid content of collagen (glycine,
proline, and hydroxyproline) was determined. Samples were extracted with 0.5 N acetic acid and precipitated with 0.9 M NaCl. Characterization tests
included those to determine the organoleptic content, pH, Fourier-transform infrared analysis, moisture content, ash content, viscosity, and Massonâ€™s
trichrome staining on collagen tissue. The collagen was further analyzed using high-performance liquid chromatography using C-18Â® column and
a fluorescence detector at 265 nm and 320 nm, acetic buffer (pH 4.2)â€“acetonitrile (55:45) as mobile phase, and optimum flow rate of 0.8 mL/min.
Results: Our findings indicated that the best method for isolating collagen was with 0.1 M NaOH expressed by average contents of glycine, proline, and
hydroxyproline in collagen which were 33.663Â±0.215%, 12.333Â±0.128%, and 11.303Â±0.354%, respectively.
Conclusion: Porcine collagen has been successfully obtained with this method.
Istanbul, Turkey: Technical Freedom and Scientific Challenges when
Transforming Collagen into Final Materials; 2013.
2. Yang H, Shu Z. The extraction of collagen protein from pig skin.
J Chem Pharm Res 2014;6:683-7.
3. Gomez-Guillen MC, Gimenez B, Lopez-Caballero ME, Montero MP.
Functional and bioactive properties of collagen and gelatin from alternative
sources: A review. Food Hydrocolloids 2011;25:1813-27.
4. Liu SH, Yang RS, al-Shaikh R, Lane JM. Collagen in tendon,
ligament, and bone healing. A current review. Clin Orthop Relat Res
5. Nithya V, Brinda P, Anand KV. Wound healing activity of Leonotis
nepetaefolia R.br, in wistar albino rats. Asian J Pharm Clin Res
6. Hossain CM, Maji HS, Chakrabority P. Hepatoprotective activity of
Lawsonia inermis linn, warm aqueous extract in carbon tetrachlorideinduced
hepatic injury in wister rats. Asian J Pharm Clin Res
7. Surya GV, Vishal GN, Gowda DV. Research and development in
wound management products: A brief review. Asian J Pharm Clin Res
8. Nagai T, Suzuki N. Isolation of collagen from fish waste material - Skin,
bone and fins. Food Chem 2000;68:277-81.
9. Lestari T. Isolation and Collagen Characterization from Thumnus
Albacares as Raw Material Pharmacy Industry. Thesis. Depok: Faculty
of Mathematics and Natural Sciences, Universitas Indonesia; 2007.
10. Rahadini IR. Isolation and Collagen Characterization of Cyprinus
carplo as Pharmaceutical Industry Raw Materials. Thesis. Depok:
Faculty of Pharmacy, Universitas Indonesia; 2007.
11. National Standardization Agency. Crude collagen from fish
scales- Quality and preparation requirement: SNI 8076-2014; 2014.
12. Kiernan JA. Histological and Histochemical Methods: Theory and
Practice. Great Britain: Pergamon PR; 1990.