# Phytochemical profile, antioxidant, antimicrobial activity, antibiofilm, cytotoxicity, and HR-LCMS of Tribulus terrestris

## TL;DR

This study explores the health benefits of Tribulus terrestris, finding it has antioxidant, antimicrobial, and anti-biofilm properties with low toxicity.

## Contribution

The study provides a comprehensive analysis of the phytochemical profile and bioactivities of Tribulus terrestris ethanolic extract.

## Key findings

- ETT showed strong antioxidant activity with IC50 values of 46.65 µg/mL (DPPH), 30.52 µg/mL (FRAP), and 41.42 µg/mL (ABTS).
- ETT exhibited antimicrobial and antibiofilm effects against MDR bacteria, with significant inhibition at 200 mg/mL.
- HR-LCMS identified diverse bioactive compounds, including alkaloids, flavonoids, and steroids.

## Abstract

Tribulus Terrestris (TT) is a Phytotherapeutic species employed in ethanopharmacological practices, stimulating spermatogenic and androgenic pathways, enhancing stamina, and promoting prophylaxis of nephrolithiasis. In this study, the focus will be on the phytochemical composition, of (TT), that contributes to antioxidant activity, antimicrobial, anti-biofilm properties.

The ethanolic extract of TT (ETT) was obtained through the Soxhlet extraction process, which was subsequently followed by rotary evaporation and lyophilization. A preliminary screening for phytochemicals was conducted, and the antioxidant activity was assessed using DPPH FRAP, and ABTS assays. Antimicrobial and antibiofilm assays were conducted. The cytotoxic effects of the ETT were evaluated utilizing L929 and HEK-293 cell lines. Scanning electron microscopy (SEM) was used to observe the morphological changes in bacteria. HR-LCMS analysis was conducted to identify its bioactive compounds.

Phytochemical screening verified the existence of phenolics, alkaloids, saponins, glycosides, flavonoids, carbohydrates, proteins, amino acids, tannins, and steroids. The phenolic concentration was measured at 7.96 µg/mL, while the flavonoid concentration was found to be 23.22 µg/ml. The ethanolic extract exhibited antioxidant activity with IC50 values of 46.65 µg/mL (DPPH), 30.52 µg/ml (FRAP), IC50 = 41.42 µg/ml (ABTS). Cytotoxicity assays revealed that ETT was non-toxic to both L929 and HEK293 cell lines across all tested concentrations. ETT has an effect against MDR bacteria. ETT markedly decreased biofilm density in a concentration-dependent manner, showing the greatest inhibition at 200 mg/ml. This was further supported by strong correlation coefficients and statistically significant p-values (p < 0.05). SEM analysis indicated considerable cellular damage in both gram-positive and gram-negative bacteria. HR-LCMS analysis identified a diverse bioactive compounds, including alkaloids, flavonoids, phenolics, saponins, tetrapyrrolic compounds, triterpenoids, glycosides, steroids, fatty acids, indanes, and methoxyphenols.

The multifaceted bioactivity of (TT) ethanolic extract, such as antioxidant activity, antimicrobial, and anti-biofilm inhibition, stems from its diverse phytochemical metabolites. This underscores their relevance in traditional ayurvedic practices and validates their.