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Dye affinity chromatography for fast and simple purification of fucoidan from marine brown algae

机译:染料亲和色谱可从海藻中快速简便地纯化岩藻依聚糖

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Fucoidan is a sulfated polysaccharide with promising pharmacological applications. Due to its medicinal properties, there is a demand for a separation technique that yields a high purification grade. Here, we present a novel purification tool for recovering fucoidan from the marine brown macroalgae Fucus vesiculosus. The developed method is based on amino-derivatized Sepabeads (R) EC-EA. The beads were modified with toluidine blue (TB), a thiazine derivative, to exploit the strong donor acceptor interactions between the cationic dye and the anionic polysaccharide. The adsorption kinetics and the binding capacity of the resin were analyzed. A Sips model was used to approximate the adsorption isotherm, resulting in a maximum capacity of 127.7 mg fucoidan per g adsorbent. Investigation of the effect of adsorption step's pH on purity and chemical structure was performed by TB and Fourier transform infra-red spectroscopy assays. Results showed that adsorption at pH 1 and 6 had negligible effects on fucoidan's chemical structure. However, purity was actually improved by 1.55- and 1.69-fold at pH 1 and 6, respectively, with an average yield of 5 g/100 g dried algae powder. In contrast, only a 1.46-fold increment was observed in fucoidan purified by the traditional method at pH 2, with a yield of 7.5 g/100 g dried algae powder. Furthermore, fucoidan purified by this method at pH 6 complies with, or even exceeds the quality of the commercially available (>= 95% pure) fucoidan (Sigma-Aldrich (R)) with respect to molecular weight and sulfur content. Therefore, dye affinity chromatography provides more advantages than the classically used techniques for fucoidan purification.
机译:岩藻依聚糖是一种硫酸化多糖,具有良好的药理应用。由于其药用特性,需要产生高纯化等级的分离技术。在这里,我们提出了一种新型的纯化工具,用于从海洋棕色大型藻类悬钩子中回收岩藻依聚糖。所开发的方法基于氨基衍生的Sepabeads(R)EC-EA。用噻嗪衍生物甲苯胺蓝(TB)对珠进行修饰,以利用阳离子染料和阴离子多糖之间的强供体受体相互作用。分析了树脂的吸附动力学和结合能力。使用Sips模型估算吸附等温线,得出最大吸附量为每克吸附剂127.7 mg岩藻依聚糖。通过TB和傅立叶变换红外光谱分析法研究了吸附步骤的pH对纯度和化学结构的影响。结果表明,在pH 1和6下吸附对岩藻依聚糖的化学结构影响可忽略不计。但是,实际上在pH 1和6下纯度分别提高了1.55和1.69倍,平均产量为5 g / 100 g干藻粉。相反,在pH 2下通过传统方法纯化的岩藻依聚糖中仅观察到1.46倍的增量,产量为7.5 g / 100 g干藻粉。此外,通过该方法在pH 6下纯化的岩藻依聚糖在分子量和硫含量方面符合或什至超过市售(纯度≥95%)岩藻依聚糖(Sigma-Aldrich)的质量。因此,染料亲和层析比岩藻依聚糖纯化的传统技术提供了更多优势。

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