Preparative isolation of flavonoid glycosides from <i>Sphaerophysa salsula</i> using hydrophilic interaction solid‐phase extraction coupled with two‐dimensional preparative liquid chromatography

In this work, a new online preparative high‐performance liquid chromatography was developed for the fast and efficient separation of complex chemical mixtures from natural products. This system integrates two chromatographic systems into an online automatic separation system using the technique of multiple trap columns with valve switching. The sample was first separated into 18 subfractions in the online preparative high‐performance liquid chromatography, and the sample eluents were then diluted and captured online on 18 trap columns by the multiple trap columns technique, respectively. Each subfraction retained on the trap column was transferred online to the separation column for the second separation. Finally, the target compounds were purified by appropriate separation conditions and multiple heart‐cutting strategies. Importantly, the system was successfully used to separate 18 high‐purity flavonoids from the crude extract of Epimedium koreanum Nakai online in one step. The entire separation time was approximately 20 h, and the structures were characterized by the high‐performance liquid chromatography quadrupole time‐of‐flight tandem mass spectrometry and nuclear magnetic resonance. This online preparative high‐performance liquid chromatography system represents an efficient and rapid separation system that has the potential for a wide array of applications in the separation of complex chemical components from natural products.

Section: Optimization Of Gradient Elution Conditionsmentioning

confidence: 99%

A novel online preparative high‐performance liquid chromatography system with the multiple trap columns‐valve switch technique for the rapid and efficient isolation of main flavonoids from Epimedium koreanum Nakai

Zhuang

Ding

et al. 2020

“…Preparative high-performance liquid chromatography (prep-HPLC) is a valuable method for isolating individual constituents from complex systems including biological samples and natural products [14][15][16][17]. This method is extensively employed in numerous areas because of its excellent column efficiency, superior separation reproducibility, online detection, and automatic control [18][19][20][21][22]. However, to protect the stationary phase from contamination and extend its service life, the crude extract cannot usually be directly injected for prep-HPLC separation.…”

Section: Introductionmentioning

confidence: 99%

Preparative isolation of arylbutanoid‐type phenol [(‐)‐rhododendrin] with peak tailing on conventional C18 column using middle chromatogram isolated gel column coupled with reversed‐phase liquid chromatography

Dang

Wang

et al. 2020

Self Cite

Reversed‐phase liquid chromatography coupled with middle chromatogram isolated gel column was employed for the efficient preparative separation of the arylbutanoid‐type phenol [(‐)‐rhododendrin] from Saxifraga tangutica. Universal C18 (XTerra C18) and XCharge C18 columns were compared for (‐)‐rhododendrin fraction analysis and preparation. Although tailing and overloading occurred on the XTerra C18 column, the positively charged reversed‐phase C18 column (XCharge C18) overcame these drawbacks, allowing for favorable separation resolution, even when loading at a on a preparative scale (3.69 mg per injection). The general separation process was as follows. First, 365.0 mg of crude (‐)‐rhododendrin was enriched from 165 g Saxifraga tangutica extract via a middle chromatogram isolated gel column. Second, separation was performed on an XTerra C18 preparative column, from which 73.8 mg of the target fraction was easily obtained. Finally, the 24.0 mg tailing peak of (‐)‐rhododendrin on XTerra C18 column was selectively purified on the XCharge C18 analytical column. These results demonstrate that the tailing nonalkaloid peaks can be effectively used for preparative isolation on XCharge C18 columns.

“…The middle chromatogram isolated (MCI) gel technique is based on the adsorption chromatography. It is widely used for crude extracts purification and natural compound enrichment [16,17]. Typically, the MCI gel technique uses an open column; however, this mode often demonstrates poor reproducibility because of the lack of controllability of the flow rate and absence of online detection [18,19].…”

Section: Introductionmentioning

confidence: 99%

A novel chromatographic separation method for rapid enrichment and isolation of novel flavonoid glycosides from Sphaerophysa salsula

Wang

Dang

Shao

et al. 2020

Self Cite

Flavonoid glycosides exist widely in medicine herbs and often used as nutraceuticals because of their excellent bioactivity and low toxicity. For accurate quality control and bioactivity assessment of Sphaerophysa salsula, a rapid and productive method to isolate flavonoid glycosides is needed. Therefore, this work reports the development of a novel comprehensive strategy based on an online middle‐pressure chromatography and preparative high‐performance liquid chromatography for rapid enrichment and separation of flavonoid glycosides from S. salsula. First, the flavonoid glycosides were enriched using an online middle‐pressure chromatographic column containing stationary middle chromatogram isolated phase. During this process, the high‐volume injection of the extracting solution was realized by an empty precolumn positioned before the main chromatographic tower. Then, the compounds were separated through preparative high‐performance liquid chromatography with Megress C18. As a result, one new flavonol 3‐O‐glycoside (2) and two known flavonol 3‐O‐glycosides (1, 3) were targetedly isolated from S. salsula. The content of compounds 1–3 in S. salsula was 0.09, 0.11, and 0.18 wt%, respectively. Comparing to traditional enrichment and separation methods, our technique offers significantly shorter sample pretreatment time as well as high reproducibility. We believe that our separation method has a strong potential to be used for the processing of other medicinal plants.