Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
J Pharm Biomed Anal. 2021 Feb 20;195:113826. doi: 10.1016/j.jpba.2020.113826. Epub 2020 Dec 7.
We report results on the structure, physicochemical characteristics and purity of chondroitin sulfate (CS) samples derived from three largely available and common biological sources such as bovine and porcine trachea and chicken keel bones with the aim to define their structural signatures. Many lots of CS produced by a manufacturer at industrial scale were characterized with a view to assess the reproducibility of the process as not controlled extractive procedures may produce final products with variable structure and biological contaminants as well as not constant clinical efficacy and safety. By using standardized source animal tissues and manufacturing procedure, highly pure CS (∼92 %) products with constant structure and characteristics were obtained. Bovine CS showed a lower molecular weight (MWw of ∼21,500 Da) than porcine (MWw of ∼26,000 Da) and chicken (MWw of ∼35,900 Da) products with a CV% of ∼2.0-7.5 and a polydispersity variability of 0.7-2.7 %. The ratio between the sulfate groups main located in position 4 and 6 of N-acetyl-galactosamine (4/6 ratio) was ∼1.70 for bovine CS versus a value of 3.60 for porcine and ∼2.70 for chicken samples with a overall charge density of 0.92-0.93 and a CV% of 2.1-2.5. The final products also showed the presence of a very low and constant content of other co-purified bio(macro)molecules (hyaluronic acid, keratan sulfate, dermatan sulfate, heparan sulfate, nucleic acids and proteins), calcium and sodium, and the absence of versican. Finally, a high reproducibility of molecular weight values, disaccharide composition, specific optical rotation and particle dimension was observed. The observed parameters are structural signatures useful to specifically identify the origin of CS and obtained by a standardized and highly reproducible manufacturing process. The compositional profile determined from this study provides a measure of the norm and range of variation in CS samples of terrestrial origin produced under standardized production protocol to which future pharmaceutical/nutraceutical final products can be compared. Moreover, the physicochemical properties including molecular weight, disaccharide composition, presence of natural contaminants and particle dimension were characterized to provide the basis of CS of high quality for application as pharmaceutical/nutraceutical active agents.
我们报告了从三种广泛存在且常见的生物来源（牛和猪气管以及鸡龙骨）中提取的硫酸软骨素（CS）样品的结构、物理化学特性和纯度的结果，目的是确定其结构特征。我们还对一家制造商在工业规模上生产的许多批次 CS 进行了表征，以评估该过程的重现性，因为非受控的提取程序可能会产生具有不同结构和生物污染物以及不同临床疗效和安全性的最终产品。通过使用标准化的原料动物组织和制造工艺，获得了具有恒定结构和特性的高纯度 CS（约 92%）产品。牛 CS 的分子量（MWw）低于猪 CS（MWw 为 26,000 Da）和鸡 CS（MWw 为 35,900 Da），CV%为 2.0-7.5，多分散性变异性为 0.7-2.7。位于 N-乙酰半乳糖胺（4/6 比）4 位和 6 位的硫酸基团的比例，对于牛 CS 约为 1.70，对于猪 CS 约为 3.60，对于鸡 CS 约为 2.70，总电荷密度为 0.92-0.93，CV%为 2.1-2.5。最终产物还显示出存在含量非常低且恒定的其他共纯化的生物（宏）分子（透明质酸、角蛋白硫酸盐、真皮素硫酸盐、肝素硫酸盐、核酸和蛋白质）、钙和钠，以及缺乏 versican。最后，观察到分子量值、二糖组成、比旋光度和粒径的高度重现性。观察到的参数是有用的结构特征，可用于特异性鉴定 CS 的来源，并通过标准化和高度可重复的制造工艺获得。从这项研究中确定的组成谱提供了在标准化生产方案下生产的陆生来源 CS 样品的标准和变化范围的度量标准，未来的药物/营养保健品最终产品可以与之进行比较。此外，还对包括分子量、二糖组成、天然污染物存在和粒径在内的物理化学性质进行了表征，为作为药物/营养保健品活性成分的高质量 CS 提供了基础。
