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April 8, 2026Methods and Protocols10 citationsOpen Access

Phenolic Compounds and Antioxidant Activity: Analytical Methods and Current Knowledge—A Review

MLMiroslav LisjakMŠMarija ŠpoljarevićJRJelena Ravlić

Key Points

  • The aim is to evaluate methodologies for extracting and analyzing phenolic compounds and their antioxidant activity.
  • Critical review of extraction and detection methodologies
  • Comparison of conventional and advanced extraction techniques
  • Evaluation of in vitro, in vivo, and ex vivo approaches
  • Discussion of spectrophotometric, electrochemical, and chromatographic detection methods
  • Lack of standardization in methodologies limits accurate assessment of antioxidant activity
  • Variations in extraction techniques impact analysis reliability
  • Combining traditional methods with innovative techniques can enhance extraction efficiency

Abstract

Phenolic compounds are plant-derived antioxidants crucial for human health and food preservation. Their bioactive potential including anti-inflammatory, antimicrobial, and anti-carcinogenic properties makes them a vital focus in nutritional, pharmaceutical, and agricultural research. This review critically evaluates the methodologies for their extraction, detection, and quantification to accurately assess antioxidant activity. Oxidative stress in biological systems and food matrices necessitates accurate analytical methodologies for assessing antioxidant behavior, which include both in vitro, in vivo and ex vivo approaches. Sample pretreatment and extraction techniques are critical for reliable analysis and vary depending on the matrix, compound polarity, and target phenolic subclass. We compare conventional extraction techniques (Soxhlet, maceration) with advanced methods like ultrasound-assisted, microwave-assisted, and supercritical fluid extraction. Detection methods reviewed include spectrophotometric assays (e.g., DPPH, FRAP, ORAC), electrochemical sensors, and chromatographic techniques (e.g., HPLC, HPLC−MS). While each method has distinct advantages, a lack of standardization remains the primary challenge, driven by variations in protocols and the vast chemical diversity of phenolics. This review underscores the critical need for integrated, standardized approaches to ensure the accurate and comparable evaluation of antioxidant activity in research and industry.

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Cite This Study

Lisjak et al. (2026) studied this question.

synapsesocial.com/papers/69d5f00974eaea4b11a79912https://doi.org/10.3390/mps9020060
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