Cui Yi, Huai Jie, Cai Yiting, Li Shuai, Zhang Weiye, Zhou Taoyulin, Song Yujie, Zhang Han, Li Shengfu, Zhuang Linwu, Zhang Junjie
Jiangsu Key Laboratory of Marine Bioresources and Environment, Jiangsu Ocean University, Lianyungang, 222005, PR China.
Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang, 222005, PR China.
Anal Bioanal Chem. 2025 Jan;417(3):583-598. doi: 10.1007/s00216-024-05669-z. Epub 2024 Nov 28.
Natural flavonoids have been shown to have many pharmacological activities. Efficient and continuous enrichment of total flavonoids with high content and low mobile phase usage from complex natural products is greatly needed at the moment. In this study, a new continuous chromatography system (CCS) with 6 zones and the 18-column dynamic tandem connection technique was developed and used to enrich total flavonoids from Epimedium koreanum Nakai (EKN). The 18 columns were divided into 6 zones, and the principle of a dynamic series of three columns was adopted for each zone to achieve continuous automatic separation and enrichment of total flavonoids under the control of a logic control valve. The CCS separation conditions were established based on single-column chromatography and a theoretical calculation model of the CCS. By means of the self-designed device and method, 485.11±3.16 g of total flavonoids were isolated from 16.2 kg of EKN. It is worth noting that the total content of 18 types of flavonoids in the samples enriched by the CCS was increased from 2.84±0.07% to 88.29±0.22%, the total recovery rate was 92.20±0.38%, and the RSD of each flavonoid was less than 5.0%. Furthermore, compared with single-column chromatography filled with the same volume of chromatography filler, the entire process saved about 2/3 of the mobile phase usage. In summary, the developed device and method could efficiently and continuously enrich total flavonoids from EKN with high-content and low mobile phase usage and would have a wide application prospect in the separation and enrichment of natural products.
天然黄酮类化合物已被证明具有多种药理活性。目前迫切需要从复杂的天然产物中高效、连续地富集高含量、低流动相用量的总黄酮。本研究开发了一种具有6个区带的新型连续色谱系统（CCS）和18柱动态串联连接技术，并用于从朝鲜淫羊藿（EKN）中富集总黄酮。18根柱子被分成6个区带，每个区带采用三柱动态串联的原理，在逻辑控制阀的控制下实现总黄酮的连续自动分离和富集。CCS的分离条件是基于单柱色谱和CCS的理论计算模型建立的。通过自行设计的装置和方法，从16.2 kg的EKN中分离得到485.11±3.16 g总黄酮。值得注意的是，经CCS富集后的样品中18种黄酮类化合物的总含量从2.84±0.07%提高到88.29±0.22%，总回收率为92.20±0.38%，各黄酮类化合物的相对标准偏差（RSD）小于5.0%。此外，与填充相同体积色谱填料的单柱色谱相比，整个过程节省了约2/3的流动相用量。综上所述，所开发的装置和方法能够高效、连续地从EKN中富集高含量、低流动相用量的总黄酮，在天然产物的分离富集中具有广阔的应用前景。
