TY - JOUR
T1 - Isolation and characteristics of bovine skin gelatin and analysis of glycine, proline, and hydroxyproline by high-performance liquid chromatography-fluorescence
AU - Khoirunnisa, Dini
AU - Harmita,
AU - Rukmana, Taufiq Indra
N1 - Publisher Copyright:
© 2018 The Authors.
PY - 2018/12/1
Y1 - 2018/12/1
N2 - Objective: This study aimed to isolate gelatin, determine its characteristics, and obtain analytical methods optimum for the determination of glycine, proline, and hydroxyproline levels in bovine gelatin. Methods: Bovine hide was hydrolyzed using 2% sodium hydroxide at an extraction temperature of 70°C for 3 h and a drying temperature of 60°C. Then, the gelatin extracts were evaluated using an organoleptic test; Fourier transform infrared analysis; pH measurement; and ash content, moisture content, and viscosity tests. Optimum analysis conditions for the determination of glycine, proline, and hydroxyproline levels in bovine gelatin using high-performance liquid chromatography with a fluorescence detector were as follows: Excitation wavelength, 265 nm; emission wavelength, 320 nm; mobile phase composition of acetic buffer: acetonitrile, 55:45; flow rate, 0.8 mL/min; C18 column with length 250 mm and inner diameter 4.6 mm; and particle size, 5 µm. Derivatization of amino acids was performed using the reagent 9-fluorenylmethoxycarbonyl chloride. Results: The results showed average levels of glycine, proline, and hydroxyproline in bovine gelatin, i.e., 25.10±0.09%, 14.28±0.11%, and 13.0±0.05%, respectively. Conclusion: The optimum conditions for bovine gelatin hydrolysis included HCl with heating for 22 h at 110°C. The hydroxyproline, glycine, and proline levels obtained for analysis in bovine gelatin samples were 13.50±0.05%, 25.11±0.09%, and 14.28±0.11%, respectively.
AB - Objective: This study aimed to isolate gelatin, determine its characteristics, and obtain analytical methods optimum for the determination of glycine, proline, and hydroxyproline levels in bovine gelatin. Methods: Bovine hide was hydrolyzed using 2% sodium hydroxide at an extraction temperature of 70°C for 3 h and a drying temperature of 60°C. Then, the gelatin extracts were evaluated using an organoleptic test; Fourier transform infrared analysis; pH measurement; and ash content, moisture content, and viscosity tests. Optimum analysis conditions for the determination of glycine, proline, and hydroxyproline levels in bovine gelatin using high-performance liquid chromatography with a fluorescence detector were as follows: Excitation wavelength, 265 nm; emission wavelength, 320 nm; mobile phase composition of acetic buffer: acetonitrile, 55:45; flow rate, 0.8 mL/min; C18 column with length 250 mm and inner diameter 4.6 mm; and particle size, 5 µm. Derivatization of amino acids was performed using the reagent 9-fluorenylmethoxycarbonyl chloride. Results: The results showed average levels of glycine, proline, and hydroxyproline in bovine gelatin, i.e., 25.10±0.09%, 14.28±0.11%, and 13.0±0.05%, respectively. Conclusion: The optimum conditions for bovine gelatin hydrolysis included HCl with heating for 22 h at 110°C. The hydroxyproline, glycine, and proline levels obtained for analysis in bovine gelatin samples were 13.50±0.05%, 25.11±0.09%, and 14.28±0.11%, respectively.
KW - 9-fluorenyl-methoxycarbonyl chloride
KW - Bovine gelatin
KW - Derivatization
KW - Glycine
KW - High-performance liquid chromatography
KW - Hydroxyproline
KW - Optimization
KW - Proline
UR - http://www.scopus.com/inward/record.url?scp=85071872603&partnerID=8YFLogxK
U2 - 10.22159/ijap.2018.v10s1.60
DO - 10.22159/ijap.2018.v10s1.60
M3 - Article
AN - SCOPUS:85071872603
SN - 0975-7058
VL - 10
SP - 269
EP - 275
JO - International Journal of Applied Pharmaceutics
JF - International Journal of Applied Pharmaceutics
IS - Special Issue 1
ER -