ISOLATION AND CHARACTERISTICS OF BOVINE SKIN GELATIN AND ANALYSIS OF GLYCINE, PROLINE, AND HYDROXYPROLINE BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHYFLUORESCENCE
DOI:
https://doi.org/10.22159/ijap.2018.v10s1.60Keywords:
Bovine gelatin, Glycine, Proline, Hydroxyproline, Derivatization, 9-fluorenyl-methoxycarbonyl chloride, High-performance liquid chromatography, OptimizationAbstract
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.
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References
Gelatin Manufacturer’s Institute of America. Standard Methods for the
Testing of Edible Gelatin. America: Gelatin Manufacturers Institute of
America, Inc; 2012.
Setiawati IH. Quality Characteristics of Chemical Physics Gelatin
Skin Red Snapper (Lutjanus sp) as Process of Acid Treatment. Bogor:
Institut Pertanian Bogor; 2009.
Mazumder R, Allamneni Y, Firdous SM, Parya H, Chowdhury AD.
Formulation, development and in-vitro release effects of ethyl cellulose
coated pectin microspheres for colon targeting. Asian J Pharm Clin Res
;6:138-44.
Farshid A, Lakshmi CS. Formulation of nanoparticles of anti-migraine drugs
triptans by coacervation method. Asian J Pharm Clin Res 2017;10:122-8.
Princely S, Saleem BN, Nandhakumar S, Dhanaraju MD. Controlled
delivery of antiretroviral drug-loaded cross-linked microspheres by
ionic gelation method. Asian J Pharm Clin Res 2016;9:264-71.
Hafidz RM, Yaakob CM, Amin I, Noorfaizan A. Chemical and functional
properties of bovine and porcine skin gelatin. Int Food Res J 2011;18:813-7.
Puspawati NM, Simpen IN, Miwada IN. Isolation of Gelatin from
Broiler Chicken Skin and Functional Group Characteristics with FTIR
Spectrophotometry. Denpasar: Department of Chemistry, Faculty of
Animal Sciences, Universitas Udayana; 2012.
Fabiani A, Versari A, Parpinello GP, Castellari M, Galassi S. Highperformance
liquid chromatographic analysis of free amino acids in
fruit juices using derivatization with 9-Fluorenylmethyl-Chloroformate.
J Chromatogr Sci 2002;40:14-8.
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