Qin Zhangyi, Du Yifan, Zhong Yue, Gu Ziyu, Zheng Qian, Zheng Yuguang, Zhang Dan, Guo Long
Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang 050200, China; International Joint Research Center on Resource Utilization and Quality Evaluation of Traditional Chinese Medicine of Hebei Province, Hebei University of Chinese Medicine, Shijiazhuang 050200, China.
Shijiazhuang Traditional Chinese Medicine Hospital, Shijiazhuang 050000, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2025 Nov 1;1265:124769. doi: 10.1016/j.jchromb.2025.124769. Epub 2025 Aug 14.
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.
粉萆薢是粉背薯蓣的干燥根茎，富含甾体皂苷。粉萆薢中存在的甾体皂苷具有很强的药理活性，尤其是在抗炎和抗高尿酸血症方面。尽管甾体皂苷被认为是粉萆薢中的主要生物活性成分，但甾体皂苷的富集和纯化过程却很少见。本研究旨在探索粉萆薢甾体皂苷的纯化工艺及其抗炎活性。首先，采用超高效液相色谱-四极杆飞行时间质谱（UPLC-Q/TOF-MS）对粉萆薢中的甾体皂苷进行表征，初步鉴定出包括原薯蓣皂苷、原纤细皂苷、伪原薯蓣皂苷、伪原纤细皂苷、薯蓣皂苷和纤细皂苷在内的6种甾体皂苷。然后，用7种大孔树脂（D-101、S-8、AB-8、ADS-7、HPD-600、HPD-100、DM130）对粉萆薢中的甾体皂苷进行纯化，选择静态吸附和解吸能力相对较强的HPD-100树脂用于粉萆薢甾体皂苷的纯化。通过静态和动态吸附和解吸实验探索HPD-100树脂的最佳纯化参数，最佳纯化条件为上样流速1.2 BV/h、上样浓度0.1 g/mL、上样体积3.5 BV、洗脱溶剂为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%）。此外，通过脂多糖（LPS）刺激的RAW264.7巨噬细胞炎症模型研究了粉萆薢粗提物和纯化甾体皂苷提取物的抗炎活性。结果表明，与粉萆薢粗提物相比，纯化后的甾体皂苷提取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树脂法从粉萆薢中纯化甾体皂苷，也为粉萆薢甾体皂苷的开发利用提供了依据。
