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February 25, 2026Biomedical Chromatography2 citationsOpen Access

Comparative Evaluation of Extraction Methods for Optimising Phenolics and Antioxidant Activity in Tribulus terrestris

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TITuğba IduğHGHikmet Gulben GucIÇIşık Çelik

Key Points

  • This research aims to evaluate the effectiveness of different extraction techniques on phenolic content and antioxidant activity in Tribulus terrestris extracts.
  • Extracts obtained from maceration, Soxhlet, ultrasound-assisted extraction (UAE), and microwave-assisted extraction (MAE) using ethanol-water solvents.
  • Total phenolic content (TPC) and total phenolic acid content (TPAC) measured through various assays.
  • Antioxidant capacity determined using DPPH, CUPRAC, and FRAP assays.
  • Phenolic acid profiles analysed by HPLC.
  • TPC values ranged from 27.66 to 37.98 mg GAE/g; TPAC ranged from 8.273 to 24.337 mg CAE/g.
  • UAE method yielded the highest DPPH antioxidant activity with an IC<sub>50</sub> of 0.1132 mg/mL.
  • CUPRAC and FRAP values varied significantly, with UAE consistently providing the most potent extracts.
  • HPLC analysis revealed eight phenolic acids, with protocatechuic acid being the most abundant.

Abstract

The medicinal plant Tribulus terrestris L. (TT) contains antioxidant and pharmacologically active phenolic compounds. However, extraction procedures can greatly affect extract yield, composition and biological activity. This study evaluated how conventional and advanced extraction methods affected TT extracts' phenolic content and antioxidant activity. TT was extracted using maceration, Soxhlet, ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) with ethanol-water solvent systems applied for different extraction times. The extracts were assessed for total phenolic content (TPC), total phenolic acid content (TPAC) and antioxidant capacity through 2,2-diphenyl-1-picrylhydrazyl (DPPH), cupric ion reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP) assays. Phenolic acid composition was analysed by HPLC. TPC values ranged between 27.66 and 37.98 mg GAE/g and TPAC between 8.273 and 24.337 mg CAE/g. Antioxidant activity (IC50) varied from 0.1132 to 0.1904 mg/mL, with UAE (80:20, 10 min) demonstrating the highest DPPH activity. CUPRAC and FRAP values extended from 97.462 to 308.769 mg TE/g and 8.917-17.291 mg TE/g, respectively, with UAE consistently yielding the most active extracts. HPLC detected eight phenolic acids, with protocatechuic acid (2.794 mg/g extract) being the most abundant, followed by p-coumaric, chlorogenic and caffeic acids. The extraction method affected the TT extracts' phenolic content and antioxidant capability. The UAE produced the highest phenolic-rich and bioactive extract. To maximise T. terrestris' phytochemical and nutraceutical potential, extraction procedures need to be optimised.

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Cite This Study

Iduğ et al. (2026) studied this question.

synapsesocial.com/papers/699e9177f5123be5ed04ef86https://doi.org/10.1002/bmc.70402
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