ISOLATION AND CHARACTERISTICS OF BOVINE SKIN GELATIN AND ANALYSIS OF GLYCINE, PROLINE, AND HYDROXYPROLINE BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHYFLUORESCENCE

  • Dini Khoirunnisa Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.
  • Harmita . Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.
  • Taufiq Indra Rukmana Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.

Abstract

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.

Keywords: Bovine gelatin, Glycine, Proline, Hydroxyproline, Derivatization, 9-fluorenyl-methoxycarbonyl chloride, High-performance liquid chromatography, Optimization.

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Khoirunnisa, D., ., H., & Rukmana, T. I. (2018). ISOLATION AND CHARACTERISTICS OF BOVINE SKIN GELATIN AND ANALYSIS OF GLYCINE, PROLINE, AND HYDROXYPROLINE BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHYFLUORESCENCE. International Journal of Applied Pharmaceutics, 10(1), 269-275. https://doi.org/10.22159/ijap.2018.v10s1.60
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