Abstract
Objective: This study aimed to validate an high-performance liquid chromatography method for analyzing undenatured Type II collagen preparationsusing a fluorescence detector.Methods: Based on the optimum analysis conditions, the compound was detected at an excitation wavelength of 255 nm and an emission wavelengthof 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. Hydroxyprolineis a compound that does not have chromophore moiety; thus, it has to be derivatized first using 9-fluorenylmetoxycarbonyl-chloride.Results: 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 detectedrange of hydroxyproline was 4–15 ppm. The limit of detection was determined to be 0.49, with an limit of quantitation of 1.64.Conclusion: Our results indicated that the average level of hydroxyproline was 98.66%, 99.12%, and 99.85%.
Highlights
Collagen is currently in high demand for various industries, including those related to food, pharmacy, and cosmetics
UC-II contains a number of amino acids, including hydroxyproline, which is a secondary amino acid derived from the proline in collagen and some plant proteins [2]
UC-II can reduce the destruction of collagen in the body, inducing anti-inflammatory activity and improving joint flexibility [5]
Summary
Collagen is currently in high demand for various industries, including those related to food, pharmacy, and cosmetics. Two types of collagen exist, namely, denatured and undenatured collagen. Undenatured collagen is termed undenatured Type II collagen (UC-II) and is derived from chicken sternum cartilage. UC-II contains a number of amino acids, including hydroxyproline, which is a secondary amino acid derived from the proline in collagen and some plant proteins [2]. Collagen has been used to address several health problems, those affecting the joints. Osteoarthritis is the most common arthritis and can affect all joints in the body [4]. UC-II can reduce the destruction of collagen in the body, inducing anti-inflammatory activity and improving joint flexibility [5]. UC-II can significantly reduce levels of circulating cytokines and associated inflammation, serving to decrease the severity of arthritis [6]
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