Open AccessISOLATION, PURIFICATION, AND CHARACTERIZATION OF PORCINE SKIN COLLAGEN: ANALYSIS OF THE GLYCINE, PROLINE, AND HYDROXYPROLINE COMPONENTS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
Author(s) -
Ayu Aditya Andayani,
Harmita Harmita,
Baitha Palanggatan Maggadani
Publication year - 2018
Publication title -
international journal of applied pharmaceutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.238
H-Index - 15
ISSN - 0975-7058
DOI - 10.22159/ijap.2018.v10s1.65
Subject(s) - hydroxyproline , proline , chromatography , chemistry , glycine , extraction (chemistry) , high performance liquid chromatography , acetic acid , imino acid , staining , amino acid , biochemistry , biology , genetics
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 testsincluded those to determine the organoleptic content, pH, Fourier-transform infrared analysis, moisture content, ash content, viscosity, and Masson’strichrome staining on collagen tissue. The collagen was further analyzed using high-performance liquid chromatography using C-18® column anda fluorescence detector at 265 nm and 320 nm, acetic buffer (pH 4.2)–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, andhydroxyproline in collagen which were 33.663±0.215%, 12.333±0.128%, and 11.303±0.354%, respectively.Conclusion: Porcine collagen has been successfully obtained with this method.