大孔树脂法纯化山药中甾体皂苷及其抗炎作用评价

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2025-08-14 DOI:10.1016/j.jchromb.2025.124769

Zhangyi Qin , Yifan Du , Yue Zhong , Ziyu Gu , Qian Zheng , Yuguang Zheng , Dan Zhang , Long Guo

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引用次数: 0

摘要

深蓝薯蓣根茎（Dioscoreae hypoglaucae rhizome， DHR）是深蓝薯蓣的干燥根茎，富含甾体皂苷。DHR中存在的甾体皂苷表现出很强的药理活性，特别是在炎症和抗高尿酸血症方面的作用。尽管甾体皂苷被认为是DHR中的主要生物活性成分，但甾体皂苷的富集和纯化工艺尚属罕见。本研究旨在探讨丹参甾体皂苷的纯化工艺及其抗炎活性。首先，采用UPLC-Q/TOF-MS对DHR中的甾体皂苷进行了表征，初步鉴定出原薯蓣皂苷、原格列西林、伪原薯蓣皂苷、伪原格列西林、薯蓣皂苷和格列西林6种甾体皂苷。然后采用7种大孔树脂（D-101、S-8、AB-8、ADS-7、HPD-600、HPD-100、DM130）对DHR中的甾体皂苷进行纯化，采用静态吸附和解吸能力较强的HPD-100树脂对DHR中的甾体皂苷进行纯化。通过静态和动态吸附与解吸实验，探索了HPD-100树脂的最佳纯化工艺参数，确定最佳纯化工艺条件为上样流速1.2 BV/h，上样浓度0.1 g/mL，上样体积3.5 BV，洗脱溶剂70% （v/v）乙醇，洗脱速度1.2 BV/h，洗脱体积3 BV。经HPD-100树脂纯化后，甾体皂苷的纯度提高了5.78倍（从6.93±0.01%提高到40.07±2.63%）。采用lps刺激的RAW264.7巨噬细胞炎症模型，研究DHR粗提物和纯化的甾体皂苷提取物的抗炎活性。结果表明，纯化后的丹参甾体皂苷提取物比粗提物具有更好的抗炎活性。本研究建立了一种可行、高效的HPD-100树脂法纯化丹参中甾体皂苷，也为丹参中甾体皂苷的开发利用提供了依据。

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Purification of steroidal saponins from dioscoreae hypoglaucae rhizoma by macroporous resin and evaluation of their anti-inflammatory effect

Dioscoreae hypoglaucae rhizome (DHR) is the dried rhizome of Dioscorea hypoglauca polibin, which is rich in steroidal saponins. The steroidal saponins present in DHR exhibit great pharmacological activities, particularly in inflammatory and anti-hyperuricemia effects. Despite steroidal saponins are recognized as primary bioactive components in DHR, the enrichment and purification processes of steroidal saponins is rare. The research aimed to explore the purification process and anti-inflammatory activity of steroidal saponins of DHR. Firstly, the steroidal saponins in DHR were characterized by UPLC-Q/TOF-MS, and six steroidal saponins including protodioscin, protogracillin, pseudoprotodioscin, pseudoprotogtacillin, dioscin and gracillin were initially identified. Then, the steroidal saponins in DHR were purified by seven types of macroporous resins (D-101, S-8, AB-8, ADS-7, HPD-600, HPD-100, DM130), and HPD-100 resin with relative great static adsorption and desorption capacities was used for purification of steroidal saponins from DHR. The optimized purification parameters of HPD-100 resin were explored by static and dynamic adsorption and desorption experiments, and optimal purification conditions were loading flow rate 1.2 BV/h, loading concentration 0.1 g/mL, loading volume 3.5 BV, elution solvent 70 % (v/v) ethanol, elution speed 1.2 BV/h and elution volume 3 BV. The purity of steroidal saponins was increased by 5.78 times (from 6.93 ± 0.01 % to 40.07 ± 2.63 %) after HPD-100 resin purification. Furthermore, the anti-inflammatory activities of the crude extracts and purified steroidal saponins extracts of DHR were investigated by LPS-stimulated RAW264.7 macrophages inflammation model. The results demonstrated the purified steroidal saponins extracts of DHR exhibited better anti-inflammatory activity compared to the crude extracts of DHR. This study conducted a feasible and efficient HPD-100 resin method for purification of steroidal saponins from DHR, and also provided evidences for the development and utilization of steroidal saponins in DHR.

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来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.