TiO₂ nanotube immobilised 5-lipoxygenase-mediated screening and isolation of anti-inflammatory active compounds from the leaves of lonicera japonica thunb

Abstract

In this work, a highly effective separation approach mediated by 5-Lipoxygenase (5-LOX) was established for screening and isolation of anti-inflammatory ingredients from leaves of Lonicera japonica Thunb. (LLJT). Using 5-LOX immobilised on TiO₂ nanotubes as a microreactor, the targeted screening was exploited by combining with HPLC-MS system. Four compounds confirmed as luteolin, luteoside, lonicerin, and isochlorogenic acid C and a fraction (M1) were screened out to be potent inhibitors of 5-LOX. Their anti-inflammatory activities were further investigated and confirmed by RAW 264.7 cells inflammation model and rat foot swelling model. Furthermore, M1 was prepared by MCI GEL CHP20P column chromatography, and further separated by Pre-HPLC. One new compound confirmed to be 5,7,3′,4′-tetrahydroxyflavone-7-O-sambubioside was first isolated from LLJT. The results provide a new method for the effective separation of active components derived from natural products.

Highlights

• A 5-LOX mediated separation method was established for isolation of anti-inflammatory compounds.

• An anti-inflammatory ingredient was separated by MCI GEL CHP20P column chromatography.

• One new compound was first isolated from leaves of Lonicera japonica Thunb.

• 5-LOX was immobilised on TiO₂ nanotubes and exploited by combining with HPLC-MS system.

• The anti-inflammatory activity of screened components was evaluated.

Keywords:

• Inflammation
• enzyme mediated isolation
• TiO₂ nanotube
• HPLC-MS
• 5-LOX Inhibitor

Author contributions

Jinhua Zhu: Project administration, Supervision, Writing-original draft, review & editing. Danyang Zhou: Methodology, Investigation. Dandan Wu: Conceptualisation, Investigation, Writing-original draft. Wei Liu: Investigation. Xiuhua Liu: Supervision.

Disclosure statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary materials.

Additional information

Funding

This research was funded by National Natural Science Foundation of China [grant number U2004133]; and the Outstanding Youth Science Fund of Henan Province [grant number 222300420031].