# Isolation and quantification of polyphenolics, exploration of antioxidant, cytotoxicity, and wound healing activities of Pithecellobium dulce (Roxb.) Benth

## TL;DR

This study explores the phytochemicals in Pithecellobium dulce, identifies active compounds, and evaluates their antioxidant, wound-healing, and cancer-fighting properties.

## Contribution

The study isolates and identifies new polyphenolic compounds from Pithecellobium dulce and evaluates their biological activities using molecular modeling.

## Key findings

- Kaempferol-3-O-rhamnoside, fisetin 3-O-rhamnoside, and alangilignoside D were isolated and identified from the ethyl acetate fraction.
- The extract showed significant antioxidant activity with an IC50 of 239.5 µg/ml in the DPPH assay.
- The extract demonstrated wound closure of 78.78% after 72 hours and cytotoxic activity against HeLa cells.

## Abstract

Pithecellobium dulce represent a valuable source of biologically active phytoconstituents. This study aimed to screen the methanolic extract for its different phytochemical classes, isolate the active principles from the ethyl acetate fraction, and conduct phenolic profiling using the HPLC-DAD technique. Additionally, the antioxidant, wound-healing, and cytotoxic activities were assessed in vitro. The molecular modelling (docking) studies are performed for the investigation of cervical and breast cancer cytotoxic activities for some isolated compounds. The extract phytochemical screening revealed the presence of flavonoids, alkaloids, anthraquinones, terpenes, sterols, tannins, saponins, carbohydrates, and reducing sugars. Kaempferol-3-O-rhamnoside (Afzelin) (1), fisetin 3-O-rhamnoside (2), and alangilignoside D (3) were isolated and identified from the ethyl acetate fraction using LC-MS, 1H and 13C NMR analysis. The methanolic extract showed total phenolics, flavonoids, tannins, and alkaloids of 51.44 mg GAE/g extract, 49.48 mg RE/g extract, 145.5 mg CE/g extract, and 18.62%, respectively. HPLC-DAD analysis enabled identifying and quantifying 13 phenolic acids and 7 flavonoids. The extract’s antioxidant activity was assessed using DPPH and ORAC assays, revealing an IC50 of 239.5 ± 8.42 µg/ml and 575.94 ± 11.30 µM TE, respectively. The extract achieved a wound closure % of 78.78 after 72 h. The cytotoxic activity was assessed against 4 cancer lines using the MTT assay, which revealed significant cytotoxic activity against the HeLa cell line, using cisplatin as a reference drug. The present investigation dealt with molecular modelling (docking) of the potent compounds showing their anticancer activities targeting cervical carcinoma HeLa cell line and breast cancer cell line MCF-7 for BCL-2 XL and EGFR. Consequently, P. dulce can be considered an effective natural antioxidant and cytotoxic agent.

The online version contains supplementary material available at 10.1038/s41598-025-32257-7.