@article{Yulianti_._Rukmana_2018, title={ISOLATION, PURIFICATION, AND CHARACTERIZATION OF BOVINE TENDON COLLAGEN AND ANALYSIS OF GLYCINE, PROLINE, AND HYDROXYPROLINE BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY-FLUORESCENCE}, volume={10}, url={https://journals.innovareacademics.in/index.php/ijap/article/view/31530}, DOI={10.22159/ijap.2018.v10s1.69}, abstractNote={<p><strong>Objective</strong>: In this study, collagen isolated from bovine tendon was purified and characterized, and the optimum conditions for analysis of glycine,<br />proline, and hydroxyproline were determined.<br /><strong>Methods</strong>: The collagen isolation process used 0.1 N NaOH as a pretreatment, 0.5 M acetic acid in the extraction, 0.9 M NaCl in the salting-out<br />step, centrifugation and dialysis for purification, and freeze-drying as the final step. The characterization of the collagen included analysis of the<br />organoleptic properties, pH, moisture content, viscosity, and ash content. A Fourier-transform infrared (FTIR) spectroscopy analysis and Casson’s<br />trichrome staining were also performed. The collagen was hydrolyzed in 6 N HCl for 24 h and derivatized using 9-fluorenylmethoxycarbonyl chloride.<br />The optimum condition was conducted from the optimal wavelength, selection of mobile phase composition, and flow rate.<br /><strong>Results</strong>: The average content was 11.867±0.20% for glycine, 33.247±0.20% for proline, and 10.51±0.23% for hydroxyproline. The optimum condition<br />analysis for collagen was achieved by high-performance liquid chromatography (HPLC) with a C18® column and a fluorescence detector (excitation:<br />265 nm and emission: 320 nm) with mobile phase acetate buffer (pH 4.2):acetonitrile (55:45), and the flow rate was 0.8 mL/min.<br /><strong>Conclusion</strong>: The collagen isolated from bovine tendon was obtained at a yield of 0.690%, and the identity was confirmed by FTIR functional group<br />analysis and Casson’s trichrome staining. The HPLC conditions using a fluorescence detector for analysis of glycine, proline, and hydroxyproline<br />concentrations in the bovine tendon collagen were optimized. The analysis of amino acids gave the average levels of 33.247±0.20% for glycine,<br />11.867±0.20% for proline, and 10.51±0.23% for hydroxyproline.</p>}, number={1}, journal={International Journal of Applied Pharmaceutics}, author={Yulianti, Dwi and ., Harmita and Rukmana, Taufiq Indra}, year={2018}, month={Dec.}, pages={311–315} }