# Metabolomic Profiling, Box–Behnken Design-Based Optimization of Ultrasonic Extraction, and Skin Anti-Aging Potential of the Green Husk of Juglans regia L. as a Sustainable Natural Waste

## TL;DR

This study explores the anti-aging properties of green husks from Juglans regia, optimizing extraction methods to maximize beneficial compounds.

## Contribution

The study introduces an optimized ultrasonic extraction method using BBD to enhance phenolic compound yield and anti-aging activity from J. regia husks.

## Key findings

- Optimal extraction conditions achieved 99.79% ethanol, 51.30°C, and 49.58 minutes for maximum phenolic content and elastase inhibition.
- Metabolomic analysis identified 41 phenolic compounds, with 24 showing increased semi-quantitative levels under optimal conditions.
- Green husks of J. regia demonstrate significant anti-aging potential due to high phenolic content and diversity.

## Abstract

The green husk of Juglans regia L. is rich in bioactive phytochemicals and exhibits various biological activities. This study aimed to investigate the skin anti-aging potential of the green husk of J. regia by determining the optimal extraction conditions using a Box–Behnken Design (BBD), targeting elastase inhibition, and by correlating variations in phenolic compounds identified through metabolomic analyses with changes in the extraction conditions. Ultrasonic-assisted extraction was employed along with natural deep eutectic solvents (NADES). Three levels of three independent variables (NADES/H2O ratio, temperature, and extraction time) were incorporated into the BBD. Phenolic compounds were determined semi-quantitatively using liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-q-TOF/MS), while elastase inhibition was evaluated by spectroscopic methods. Quadratic response surface models were proposed based on the BBD model adequacy test using multiple regression analysis. It was determined that the optimum conditions for maximizing phenolic content and elastase inhibition were 99.79% ethanol, 51.30 °C temperature, and 49.58 min, in which case the desirability score was 1. Metabolomic analysis identified 41 phenolic compounds across 27 ontological groups, with 24 compounds showing a semi-quantitative increase. Consequently, the waste green husk of J. regia demonstrated enhanced anti-aging potential due to the higher content and diversity of phenolic compounds.