TY - JOUR AU - Maggadani, Baitha Palanggatan AU - ., Harmita AU - Lubnan, Milza PY - 2018/12/20 Y2 - 2024/03/28 TI - VALIDATION METHODS FOR THE ANALYSIS OF HYDROXYPROLINE FROM COLLAGEN UNDENATURED TYPE II COLLAGEN USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY FLUORESCENCE JF - International Journal of Applied Pharmaceutics JA - Int J App Pharm VL - 10 IS - 1 SE - Original Article(s) DO - 10.22159/ijap.2018.v10s1.89 UR - https://journals.innovareacademics.in/index.php/ijap/article/view/31575 SP - 403-406 AB - <p><strong>Objective</strong>: This study aimed to validate an high-performance liquid chromatography method for analyzing undenatured Type II collagen preparations<br />using a fluorescence detector.<br /><strong>Methods</strong>: Based on the optimum analysis conditions, the compound was detected at an excitation wavelength of 255 nm and an emission wavelength<br />of 320 nm. The optimum mobile phase was determined to be acetate (pH 4.2) and acetonitrile (60:40) with a flow rate of 1.0 ml/min. Hydroxyproline<br />is a compound that does not have chromophore moiety; thus, it has to be derivatized first using 9-fluorenylmetoxycarbonyl-chloride.<br /><strong>Results</strong>: The developed method was validated with linearity and an equation of y=3,249,704 x+141,945,072, with a value of r=0.9994. The detected<br />range of hydroxyproline was 4–15 ppm. The limit of detection was determined to be 0.49, with an limit of quantitation of 1.64.<br /><strong>Conclusion</strong>: Our results indicated that the average level of hydroxyproline was 98.66%, 99.12%, and 99.85%.</p> ER -