Fractionation, HRLC-MS assisted phytochemical profiling, quantification, and evaluation of the chronic wound healing potential of ethanolic extract of Ardisia solanacea (Poir.) Roxb.

Abstract

Background

Wound healing is a complex, multi-stage biological process involving coordinated events such as haemostasis, inflammation, proliferation, and remodelling. When these stages fail to proceed in a timely and orderly manner, chronic wounds can develop. Herbal medicines, contain a blend of phytoconstituents, can act on multiple phases of wound healing and, being naturally derived, may have fewer side effects. Ardisia solanacea (Poir.) Roxb., a member of the genus Ardisia, has been used in traditional medicine for its wide-ranging activities, including antioxidant, anti-inflammatory, antimicrobial, antibacterial, and antidiabetic, etc. The present study aims to analyze and quantify the phytochemicals in the ethanolic extract from A. solanacea using various chromatographic techniques, and to assess its in vivo chronic wound-healing potential.

Methods

In vitro antioxidant potential of ethanolic extract was evaluated by metal chelating assay. The phytochemical profiling was conducted through chromatographic techniques such as flash chromatography, HRLC-MS, and HPTLC. The chronic wound healing potential of A. solanacea was evaluated using an excision wound model, where diabetes mellitus was induced in wistar albino rats using alloxan monohydrate (120 mg/kg).

Results

The ethanolic extract of A. solanacea showed an IC₅₀ value of 65.41 ± 0.80 µg/mL, compared to 61.09 ± 1.12 µg/mL for BHT (standard). Phytochemical profiling through flash chromatography isolated 48 fractions, with fraction no. 12 being further analyzed by HRLC-MS, which identified 26 secondary metabolites. Among these were newly discovered compounds, including sinensetin, along with several flavonoid, phenolic, and alkaloid derivatives. HPTLC was employed to quantify sinensetin in both the ethanolic extract and the emulgel formulation. Pharmacological evaluation in wistar albino rats revealed that the formulated Ardisia solanacea emulgel, 10 % w/w has significant chronic wound healing potential, achieving 93.04 % ± 0.32 wound contraction by the 21st day, and a significantly shortened epithelization period (p < 0.05) compared to diabetic control group.

Conclusion

To the best of our knowledge, this is the first study to demonstrate the chronic wound healing potential of the ethanolic extract of A. solanacea through excision wound model. This study supports the traditional use of A. solanacea for treating wounds and provides the scientific evidence confirming its chronic wound healing potential.