In vitro Collagenase Inhibition and IC₅₀ Benchmarking of Woodfordia fruticosa Extract

Collagenase (MMP 1) is a key enzyme in dermal collagen degradation and wrinkle formation. Natural inhibitors from medicinal plants offer promise for anti-aging therapy, but standardized in vitro validations remain scarce. To evaluate the collagenase inhibitory activity of Woodfordia fruticosa flower extract and benchmark its IC₅₀ against published plant-based inhibitors under comparable assay conditions. Flowers of W. fruticosa were collected from the Sahyadri ranges, authenticated, and extracted with ethanol using accelerated solvent extraction. A microplate-based endpoint assay with FALGPA substrate was used to measure collagenase inhibition at 335 nm. Extract dilutions (3.12–400 μg/mL) were assayed in triplicate, and IC₅₀ was calculated by nonlinear regression. LC MS confirmed the presence of vitexin. Benchmarking was performed with reported IC₅₀ values for Curcuma longa, Camellia sinensis, and epigallocatechin gallate (EGCG). The ethanol extract of W. fruticosa showed dose-dependent inhibition, with an IC₅₀ of 131.04 μg/mL. This potency exceeded that of C. longa (~200 μg/mL) and C. sinensis (~250 μg/mL) extracts, and comparable with reference polyphenols under the same assay conditions. LC MS confirmed vitexin as a major flavonoid constituent, supporting its role in activity. W. fruticosa extract demonstrates significant collagenase inhibition and compares favorably with established botanicals. The results provide reproducible biochemical evidence supporting its development as a natural anti-aging agent, and establish a standardized reference point for future mechanistic and formulation studies.