ISOLATION, PURIFICATION, AND CHARACTERIZATION OF PORCINE SKIN COLLAGEN: ANALYSIS OF THE GLYCINE, PROLINE, AND HYDROXYPROLINE COMPONENTS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
Keywords:Amino acid, Derivatization, Fluorescence, Glycine, High-performance liquid chromatography, Hydroxyproline, Porcine skin collagen, Praline
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 Masson'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.
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