色谱-粘度法测定分子量：以紫茎泽兰新多糖为例

IF 2.4 3区化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Carbohydrate Research Pub Date : 2025-05-20 DOI:10.1016/j.carres.2025.109542

Sushil Bhattarai, Vineet Kumar

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引用次数: 0

摘要

天然多糖的分子量分析是各种营养、制药、生物、化学和其他工业应用所必需的。本研究除了对紫茎泽兰的组成单糖进行化学成分分析外，还对紫茎泽兰多糖的分子量进行了独立的测定。通过反复沉淀、离心、透析、层析纯化等工艺对该多糖进行分离纯化，得到该多糖的产率为0.62±0.08%。为了分析构成多糖的单糖，首先对其进行水解，然后将水解产物还原，并通过乙酰化衍生成相应的糖醇乙酸酯。利用已知分子量的葡聚糖标准物，用HPGPC法测定分子量、平均分子量和数平均分子量。粘度的平均分子量是用乌伯洛德稀释粘度计和Mark-Houwink方程测定的特性粘度。GC-MS/FID分析表明，其单糖组成比例为1.61:1.00:0.25:5.89:3.24 M，分别为l-阿拉伯糖、d-木糖、d-甘露糖、d-葡萄糖和d-半乳糖。测定其分子量分别为4.05 × 105 Da （MW）、2.28 × 105 Da （MN）和3.59 × 105 Da （MN）。该多糖具有多分散性，PDI值为1.793。这种谨慎的研究对于第三世界国家和本科生实验室是有用的，它们需要基于分子量的引导筛选，用于各种应用，但在实验室中没有HPGPC。结果表明，一个简单的玻璃ubbelode稀释粘度计满足了分子量测定的目的。本文首次报道了紫参叶多糖的分子量和单糖组成。

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Molecular weight determination by Chromatography and Viscometry: A case study using novel polysaccharide of Eupatorium adenophorum L

Molecular weight analysis of natural polysaccharides is necessary for diverse nutraceutical, pharmaceutical, biological, chemical and other industrial applications. The present study focuses on the determination of the molecular weight of polysaccharide from Eupatorium adenophorum using HP-GPC and Viscometry independently, apart from the chemical composition of constituent monosaccharides. The novel polysaccharide was isolated from leaves of the species, followed by purification using repetitive precipitation, centrifugation, dialysis and chromatographic purification to obtain the polysaccharide in 0.62 ± 0.08 % yield. To analyze monosaccharides constituting polysaccharide, it was hydrolyzed, followed by reduction of hydrolysates and derivatization to corresponding alditol acetates by acetylation. Weight average and number average molecular weights were determined by chromatography (HPGPC) with the help of known MW dextran standards. The viscosity average molecular weight was determined by intrinsic viscosity measurement using the Ubbelohde dilution viscometer and applying the Mark-Houwink equation. GC-MS/FID analysis indicated the presence of l-arabinose, d-xylose, d-mannose, d-glucose and d-galactose as the constituent monosaccharides in 1.61:1.00:0.25:5.89:3.24 M ratios. The different molecular weights were determined to be 4.05 × 10⁵ Da (M_W), 2.28 × 10⁵ Da (M_N) and 3.59 × 10⁵ Da (M_ƞ). The polysaccharide has a polydisperse nature with PDI value of 1.793. Such prudent studies are useful for third-world countries and undergraduate laboratories requiring the guided molecular weight-based screening for diverse applications, but do not have HPGPC in the laboratory. Results indicate that a simple glass Ubbelohde Dilution Viscometer fulfils the goal of molecular weight determination. The molecular weight and monosaccharide composition of E. adenophorum leaf polysaccharide have been reported for the first time.

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来源期刊

Carbohydrate Research 化学-生化与分子生物学

CiteScore

5.00

自引率

3.20%

发文量

183

审稿时长

3.6 weeks

期刊介绍： Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects. Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence. Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".