天然产物研究与开发 ›› 2025, Vol. 37 ›› Issue (1): 95-103.doi: 10.16333/j.1001-6880.2025.1.011 cstr: 32307.14.1001-6880.2025.1.011

• 开发研究 • 上一篇    下一篇

霍山石斛叶黄酮提取纯化工艺优化及其成分分析

韩   硕1,2,李成网3,4,王厉梅6,宫   鹏2*,刘增辉3,4,5*   

  1. 1安徽医科大学药学院;2安徽医科大学第一附属医院药剂科,合肥 230022;3安徽省医学科学研究院;4安徽省医学高等专科学校,合肥 230601;5安徽省祁门县蛇伤研究所,祁门 245600;6浙江康牧药业有限公司,绍兴 312400
  • 出版日期:2025-01-22 发布日期:2025-01-22
  • 基金资助:
    2022年度高等学校省级质量工程项目(2022kcsz149);安徽省高校自然科学基金重点项目(2024AH050851);安徽省科技重大专项(202203c08020004);安徽省中医药学会中医药科研项目(2024ZYYXH175)

Optimization of extraction and purification process of Dendrobium huoshanense C.Z.Tang et S.J.Cheng leaf flavonoids and its component analysis

HAN Shuo1,2,LI Cheng-wang3,4,WANG Li-mei6,GONG Peng2*,LIU Zeng-hui3,4,5*   

  1. 1College of Pharmacy,Anhui Medical University;2The First Affiliated Hospital of Anhui Medical University,Hefei 230022,China;3Anhui Academy of Medical Sciences;4Anhui Medical College,Hefei 230601,China;5Qimen Institute of Snakebite of Anhui Province,Qimen 245600,China;6Zhejiang Kangmu Pharmaceutical Co.,Ltd.,Shaoxing 312400,China
  • Online:2025-01-22 Published:2025-01-22

摘要:

以霍山石斛(Dendrobium huoshanense C.Z.Tang et S.J.Cheng)叶为材料,建立石斛叶黄酮的提取及纯化工艺,并分析其主要黄酮类成分。本研究以乙醇浓度、料液比、提取温度和回流时间为考察因素,以夏佛塔苷得率为评价指标,正交试验法建立霍山石斛叶黄酮最佳提取工艺;并从树脂选择、上样体积优化和洗脱液选择的角度,考察夏佛塔苷回收率,建立最佳纯化工艺;利用UPLC-Q-Exactive Orbitrap MS对提取物中主要黄酮类成分进行分析。结果表明,霍山石斛叶黄酮的最佳提取工艺为:乙醇体积分数70%、料液比1∶30、提取温度90 ℃、提取时间30 min。此条件下,夏佛塔苷得率为0.348 2%。以夏佛塔苷含量为85 μg/mL上样液进行纯化时,最佳纯化工艺为:采用AB-8型大孔吸附树脂为纯化材料,上样体积7 BV,洗脱液为6 BV 50%乙醇,夏佛塔苷回收率为96.68%,纯化后所得干粉中夏佛塔苷含量为5.11%,较纯化前含量提高355%。从提取物中分析鉴定出33个黄酮类化合物,其中24个是首次在霍山石斛中发现。以上结果表明,该提取纯化工艺稳定可行,可用于霍山石斛叶中黄酮类成分的提取纯化,所得黄酮含量较高。

关键词: 霍山石斛叶, 黄酮, UPLC-Q-Exactive Orbitrap MS, 夏佛塔苷, 提取纯化工艺

Abstract:

This study aims to establish a process applicable to the extraction and purification of flavonoids from the leaves of Dendrobium huoshanense C.Z.Tang et S.J.Cheng and to analyse its main flavonoid components.In this study,ethanol volume fraction,solid-liquid ratio,temperature and reflux time were selected as potential influencing factors in the design of an orthogonal experiment to establish an optimal extraction process by comparing the yield of shaftoside.We also investigated the influence of various purification resins,sample volumes and eluent variables on shaftoside recovery in order to develop an optimal purification process.Furthermore,UPLC-Q-Exactive Orbitrap MS was employed to investigate the primary chemical components of the flavonoids present in the extract.The results demonstrated that the optimal extraction process for D. huoshanense leaf flavonoids was achieved with a 70% ethanol solution,a solid to liquid ratio of 1∶30,an extraction temperature of 90 ℃,and an extraction time of 30 minutes.And the yield of shaftoside was 0.348 2% under these conditions.The optimal purification procedure was as follows:AB-8 resin as the purification material,7 BV sample (shaftoside content was 85 μg/mL),and 6 BV of 50% ethanol as the eluent.The recovery of shaftoside under these condition was 96.68%.The content of shaftoside in the dry powder obtained after purification was 5.11%,representing a significant increase of 355% in comparison to the unpurified sample.33 flavonoids were identified in the extract,of which 24 were found for the first time in D. huoshanense.The results indicated that the extraction and purification process was stable and feasible,and can be used for the extraction and purification of flavonoid components in D. huoshanense leaves to obtain high quality flavonoids.

Key words: Dendrobium huoshanense leaves, flavonoid, UPLC-Q-Exactive Orbitrap MS, schaftoside, extraction and purification process

中图分类号:  R284.2