Green Preservation Strategies: The Role of Essential Oils in Sustainable Food Preservatives

Abstract

Essential oils (EOs) have gained increasing attention as natural alternatives to synthetic food preservatives due to their broad-spectrum antimicrobial, antioxidant, and antigenotoxic properties. Derived from aromatic plants, EOs possess complex chemical compositions rich in bioactive compounds such as terpenes, phenolics, and aldehydes, which contribute to their effectiveness against foodborne pathogens, oxidative spoilage, and genotoxic contaminants. This review provides a comprehensive examination of the potential of EOs in food preservation, highlighting their mechanisms of action, including membrane disruption, efflux pump inhibition, and reactive oxygen species scavenging. Standard assays such as disk diffusion, MIC/MBC, time-kill kinetics, and comet and micronucleus tests are discussed as tools for evaluating efficacy and safety. Additionally, the use of EOs in diverse food matrices and the reduction in reliance on synthetic additives support cleaner-label products and improved consumer health. The review also examines the sustainability outlook, highlighting the potential for extracting EOs from agricultural byproducts, their integration into green food processing technologies, and alignment with the circular economy and the Sustainable Development Goals. Despite promising results, challenges remain in terms of sensory impact, regulatory approval, and dose optimization. Overall, EOs represent a multifunctional and sustainable solution for modern food preservation systems.

1. Introduction

Modern food systems face increasing pressure to extend shelf life and ensure microbiological safety while responding to consumer demand for natural, minimally processed products [1]. Synthetic preservatives, once the industry standard, are now viewed with skepticism due to growing concerns over potential health and environmental impacts. This shift has prompted the search for multifunctional, natural alternatives that can maintain food quality while ensuring safety and sustainability.

Essential oils (EOs), volatile plant-derived compounds rich in bioactive constituents such as terpenes, phenolics, and aldehydes, have emerged as promising candidates. Their antimicrobial and antioxidant properties are well documented; however, their potential as antigenotoxic agents—capable of mitigating DNA damage caused by foodborne contaminants—remains underexplored in food preservation contexts [2,3]. EOs are a specific class of aromatic products that also encompass extracts produced by other techniques, such as cold pressing, solvent extraction, or supercritical CO₂ extraction [4]. However, not all these methods yield true essential oils, and this distinction is critical for understanding their functionality, regulatory status, and application in food systems.

This review adopts a multifunctional lens, synthesizing current evidence on the antimicrobial, antioxidant, and antigenotoxic activities of EOs while addressing sustainability challenges and regulatory considerations. Unlike previous reviews, which focus primarily on individual functions or compositional analyses, we highlight EOs as integrated tools for developing clean-label products and aligning food systems with green processing technologies and circular economy principles. This systems-level perspective provides a strategic roadmap for future innovation in natural food preservation.

2. Essential Oils: Composition, Extraction, and Regulatory Aspects

2.1. Chemical Composition and Active Constituents

EOs are highly complex mixtures of volatile, low molecular-weight compounds synthesized by aromatic plants as part of their natural defense and communication mechanisms. These mixtures typically consist of 20 to 80 components, although a few major constituents often account for up to 85–90% of the oil’s composition [5,6].

The main classes of bioactive constituents found in EOs include the following:

• Terpenes (e.g., limonene, pinene, sabinene);
• Terpenoids (oxygenated terpenes, e.g., linalool, geraniol, thymol, carvacrol);
• Phenolic compounds (e.g., eugenol, thymol);
• Aldehydes (e.g., citral, cinnamaldehyde);
• Ketones (e.g., menthone, carvone);
• Alcohols (e.g., borneol, terpineol);
• Esters (e.g., linalyl acetate).

These compounds contribute to the organoleptic properties (aroma and flavor) of EOs and are primarily responsible for their biological activities, including antioxidant, antimicrobial, and antigenotoxic effects. Their mechanisms of action are diverse and often synergistic. For instance, phenols such as thymol and carvacrol can disrupt microbial membranes and generate oxidative stress in pathogens [7]. In contrast, compounds such as eugenol and linalool exhibit both radical-scavenging and anti-inflammatory properties, thereby contributing to food safety and extending shelf life [8,9].

The biosynthesis of EO constituents depends on the plant’s genetics, phenology, and environmental conditions, including climate, soil, harvesting time, and geographical origin. This variability directly impacts the efficacy and reproducibility of EOs as food preservatives. Therefore, chemotype identification—based on dominant compounds—is a critical quality control step, especially for regulatory and formulation purposes. For example, Thymus vulgaris may exist as a thymol- or linalool-dominant chemotype, with distinct antimicrobial spectra [10].

Modern analytical techniques such as gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) spectroscopy are essential for qualitative and quantitative profiling of EOs. GC-MS, in particular, is the gold standard for EO characterization due to its ability to separate and identify individual components with high precision and reproducibility [11].

Furthermore, it is increasingly important to assess batch-to-batch variability, especially when considering industrial-scale use in food systems. Standardized characterization allows producers to ensure consistent efficacy, particularly when EOs are used in active packaging, emulsions, or nanoencapsulated forms.

Finally, while minor components are often overlooked, they may contribute significantly to synergistic effects or mitigate the adverse sensory impacts of major constituents. Thus, a full-spectrum approach to EO composition analysis is encouraged, especially in the context of food additives, where both bioactivity and organoleptic balance are critical [12].

2.2. Extraction Methods and Impact on Sustainability

The extraction method used to obtain EOs plays a pivotal role in determining the yield, chemical composition, biological activity, and environmental footprint of the final product. Traditionally, hydrodistillation and steam distillation have been the most commonly used techniques [13]. However, growing environmental concerns and the demand for green chemistry principles in food and cosmetic industries have driven the development of more sustainable, efficient, and selective extraction technologies.

2.2.1. Conventional Extraction Techniques

Steam Distillation is widely used for commercial EO production. It involves passing steam through plant material, volatilizing the EOs, and condensing it back into liquid form. While reliable and scalable, this method often requires long processing times and high energy input, which may degrade heat-sensitive compounds and result in a lower yield of oxygenated or minor constituents [14].

Hydrodistillation is similar but involves immersing plant material in water. It may lead to hydrolysis or thermal degradation of certain compounds and is less favorable for large-scale or industrial applications.

These conventional methods, although time-tested, are increasingly scrutinized for their environmental impact and inefficient use of water and energy resources.

2.2.2. Green and Advanced Extraction Technologies

To improve extraction sustainability and selectivity, several non-conventional methods have been developed, including the following:

• Supercritical Fluid Extraction (SFE): Uses supercritical CO₂ as a solvent, offering high selectivity, mild temperatures, and minimal solvent residues in the final product. SFE allows the extraction of thermolabile and non-polar compounds with lower energy consumption and no water waste, aligning well with eco-friendly processing goals [15]. However, high equipment costs can be a barrier for small producers.
• Microwave-Assisted Extraction (MAE): Utilizes microwave energy to heat the plant matrix rapidly and efficiently. MAE reduces extraction time and solvent use and can enhance the recovery of specific bioactives [16]. It is increasingly recognized as a sustainable technique for industrial-scale EO recovery.
• Ultrasound-Assisted Extraction (UAE): Relies on acoustic cavitation to disrupt plant cells and facilitate the release of EOs. The UAE enhances yield, reduces energy consumption, and shortens extraction time compared to classical techniques [17]. It is particularly effective for delicate aromatic herbs.
• Ohmic Heating: An emerging technique where plant material is heated via electrical current. It enables rapid, uniform heating and shows promise in EO extraction, particularly when combined with hydrodistillation, thereby improving energy efficiency [18].

Each of these techniques can be optimized to meet specific industrial needs, and lifecycle assessments indicate that they have lower environmental footprints compared to traditional approaches.

Other Industrial Extraction Methods

In addition to the methods discussed above, several other extraction techniques are employed in the aroma and flavor industries to recover volatile and semi-volatile compounds from plant matrices:

• Solvent Extraction: This method uses polar (e.g., ethanol, methanol) or non-polar (e.g., hexane, petroleum ether) solvents to extract aromatic compounds. It is particularly effective for delicate flowers (e.g., jasmine, tuberose) that do not tolerate high heat. However, solvent residues, potential toxicity, and environmental concerns limit its use in food-grade applications unless followed by rigorous purification [19].
• Enzyme-Assisted Extraction (EAE): Enzymes such as cellulases or pectinases are used to degrade plant cell walls, enhancing the release of essential oil constituents and other bioactives. EAE is considered mild and selective, preserving thermolabile compounds while improving yield and extraction efficiency [20,21].
• Pressurized Hot Water Extraction (PHWE): Also known as subcritical water extraction, PHWE uses water at elevated temperatures (100–374 °C) and pressure to extract polar and semi-polar compounds. It offers a greener alternative to organic solvents and can be used to obtain antioxidant-rich extracts with minimal degradation of sensitive molecules [22,23].

These techniques differ significantly in their selectivity, environmental impact, and suitability for various raw materials. Importantly, the choice of extraction method is a key determinant of the chemical composition and, consequently, the bioactivity, sensory profile, and regulatory status of the resulting aromatic product. Method optimization is therefore crucial for tailoring extracts to specific food preservation applications.

2.2.3. Waste Valorization and Circular Economy Approaches

A critical innovation in sustainable EO production involves sourcing raw materials from agro-industrial residues and food waste, such as citrus peels, grape pomace, and herb trimmings. These by-products, often discarded or underutilized, are rich in volatile compounds and can be transformed into value-added products through EO extraction [24]. This approach supports waste minimization, resource efficiency, and bioeconomy principles, while also reducing production costs and promoting environmental stewardship.

For example, citrus peel waste from juice production has been successfully used to extract limonene-rich EOs with antioxidant and antimicrobial properties applicable in food preservation [25,26]. Similarly, rosemary and oregano waste from herbal industries can be processed into high-value antimicrobial agents for edible coatings or active packaging [27,28].

2.3. Regulatory Status of EOs in Food Systems

The regulatory framework governing the use of EOs in food systems is complex and varies significantly across countries and regions. As EOs are multifunctional substances used for flavoring, preservation, and even therapeutic purposes, their legal status depends on their intended use, composition, and safety profile. Ensuring regulatory compliance is crucial for the adoption of natural preservatives, particularly in commercial and export-oriented food production.

2.3.1. GRAS Status and Food Additive Approvals

Despite GRAS status in the United States, the lack of harmonized global standards hampers industrial scalability. The Food and Drug Administration (FDA) recognizes many EOs as GRAS when used by good manufacturing practices. Examples include clove oil, cinnamon oil, lemon oil, and thyme oil, which are permitted as flavoring agents and have been documented to have antimicrobial properties [29,30].

However, GRAS status for flavoring purposes does not automatically confer approval for use as a preservative or antimicrobial, which falls under different regulatory scrutiny [31]. Manufacturers must demonstrate efficacy, stability, and toxicological safety for the specific application, including potential migration into food when EOs are used in packaging materials.

2.3.2. European Union Regulations

In the European Union (EU), EOs used in food must comply with Regulation (EC) No 1334/2008 on flavorings and food ingredients with flavoring properties. EOs considered as flavoring substances are generally allowed if they meet safety requirements established by the European Food Safety Authority (EFSA) [32].

However, when EOs are used for technological purposes, such as microbial inhibition, they may be subject to Regulation (EC) No 1333/2008 on food additives. In this case, a food additive code (E-number) and an assessment of acceptable daily intake (ADI) may be required. Currently, only a few EO constituents are registered as food preservatives (e.g., eugenol, thymol), and whole EOs used for preservation remain in a legal grey zone in many EU member states [33].

Additionally, under Regulation (EU) No 10/2011, EOs used in active packaging systems must not migrate into food in concentrations that exceed established limits unless authorized [34].

2.3.3. Codex Alimentarius and International Guidelines

The Codex Alimentarius, developed by the FAO and WHO, offers global guidelines for food additives and flavorings. Although not legally binding, Codex standards have an influence on national policies and trade regulations. The Codex General Standard for Food Additives (GSFA) lists several EO-derived substances approved for use in flavoring and preservation purposes, including citral, cinnamaldehyde, and limonene [35,36].

Nevertheless, Codex guidelines do not currently provide comprehensive regulation for the use of whole EOs as preservatives, which limits harmonization and complicates international marketing of EO-based food products [37].

2.3.4. Safety Assessments and Toxicological Requirements

For regulatory approval, particularly for novel food preservative claims, EOs must undergo toxicological risk assessments, including the following:

• Acute and chronic toxicity;
• Genotoxicity and antigenotoxicity;
• Allergenicity and potential sensitization;
• ADI and exposure modeling [38,39].

Despite being natural, some EO constituents may exhibit cytotoxic or pro-oxidant effects at high concentrations, especially in sensitive populations such as children or pregnant women. Therefore, optimization and encapsulation technologies are increasingly explored to enhance efficacy and reduce potential risks [40,41].

Regulatory agencies also consider the variability in EO composition resulting from differences in plant source, extraction method, and seasonal variations. Hence, standardization and batch testing are essential to comply with labeling and safety requirements [42,43].

2.3.5. Organic Certification and Clean-Label Trends

The use of EOs aligns with the clean-label movement and organic certification schemes, which favor minimally processed, natural ingredients [44,45]. However, to qualify for labels such as USDA Organic or EU Organic, the plant source and extraction process must also be certified. Solvent residues, contamination, or adulteration can disqualify EOs from organic or natural claims, thereby affecting their marketability [46,47].

These certifications can enhance consumer trust and broaden the appeal of EO-based preservatives, particularly in the functional food and health-conscious markets.

Table 1. Summary of regulatory status of essential oils in food systems by region.

Region	Regulatory Body	EOs Approved As	Key Requirements	Examples
United States	FDA (GRAS Program)	Flavorings (GRAS status); not all approved as preservatives	Must comply with GRAS evaluation for specific uses; safety and efficacy data needed for preservative claims	Clove EO, cinnamon EO, lemon EO
European Union	EFSA/EC	Flavorings (Reg. 1334/2008); Additives (Reg. 1333/2008)	Safety assessments, ADI values, E-number registration for additive use	Thymol, eugenol (as components)
Codex Alimentarius (Global)	FAO/WHO (Codex GSFA)	Individual EO compounds as additives or flavorings	Non-binding but serves as reference for trade and national regulation	Citral, limonene, cinnamaldehyde
China	NHC	Limited use as food flavors	Approval required for food additive use; strictly regulated	Star anise EO, orange EO
Japan	MHLW	Food flavorings under the positive list	Additive use must be authorized; generally, restrictive	Menthol, peppermint EO

ADI: Acceptable Daily Intake; EC: European Community; EFSA: European Food Safety Authority; FAO: Food and Agriculture Organization; FDA: Food and Drug Administration; GRAS: Generally Recognized As Safe; GSFA: General Standard for Food Additives; MHLW: Ministry of Health, Labor and Welfare; NHC: National Health Commission; WHO: World Health Organization.

summarizes key regulatory distinctions for EO use across major markets. While many essential oils are approved as flavorings, their use as preservatives is often subject to additional scrutiny. The lack of global harmonization poses challenges for international product development, highlighting the need for region-specific safety data, standardized formulations, and transparent labeling practices.

2.4. Sustainability Considerations in Essential Oil Production and Application

EOs are complex mixtures of volatile secondary metabolites whose composition and biological activity can vary significantly due to factors such as plant genetics, geographic origin, cultivation practices, harvest time, and extraction method [48]. This intrinsic variability poses significant challenges for their standardization, reproducibility, and regulatory approval, mainly when EOs are intended for use as food preservatives. In addition to compositional variability and quality control challenges, sustainability in EO applications must also address the environmental impact of extraction processes, regulatory alignment with sustainability goals, and the adoption of circular economy practices. These considerations are essential for ensuring that EO-based food preservatives meet both ecological and legal standards in modern food systems.

To ensure safety, efficacy, and regulatory compliance, robust quality control systems and standardization protocols are crucial. These include chemical profiling, biological activity assays, and contamination checks, which collectively contribute to the reproducibility of EO-based food applications and foster consumer trust in natural preservatives.

2.4.1. Importance of Standardization

Standardization refers to the process of defining consistent chemical and biological characteristics of EOs across different production batches. This is crucial because of the following:

• Minor variations in composition can alter antimicrobial, antioxidant, or sensory properties.
• Regulatory bodies require transparent specification sheets for food-grade materials.
• Food manufacturers depend on predictable functionality and stability in product formulation.
• The reproducibility of scientific data depends on the use of well-characterized materials.

Lack of standardization may lead to inconsistent preservation efficacy, unexpected toxicity, or loss of organoleptic quality in food applications [49,50].

2.4.2. Sustainable Extraction and Environmental Metrics

Green extraction techniques, such as supercritical CO₂ extraction, microwave-assisted extraction, and ultrasound-assisted extraction, offer reduced energy and water usage, lower emissions, and better preservation of bioactives compared to traditional methods like hydrodistillation or steam distillation [51]. These methods support life cycle efficiency and align with sustainable processing principles.

In parallel, sustainability metrics—including water and energy consumption, land use, carbon footprint, and solvent toxicity—are increasingly used to assess EO production [52]. Eco-certification schemes and traceability systems are recommended to validate environmental claims, particularly when EOs are used in food products marketed under clean-label or organic categories [53].

2.4.3. Regulatory Support for Sustainability

Recent regulatory frameworks are progressively incorporating sustainability into food additive evaluation. The European Green Deal and Farm to Fork Strategy specifically promote the use of natural, plant-based preservatives to reduce chemical dependency in food systems [54]. These trends encourage the development and use of EOs, provided they meet established safety and efficacy standards. Regulatory bodies are beginning to consider environmental performance—such as renewable sourcing and low-impact processing—alongside traditional toxicological and functional data.

2.4.4. Chemical Characterization Techniques

The primary tools for essential oil quality control are chromatographic and spectrometric techniques that provide qualitative and quantitative data on EO composition:

• Gas chromatography coupled with mass spectrometry (GC–MS) is the gold standard for identifying and quantifying volatile components. It enables detection of both major (e.g., thymol, eugenol, citral) and minor constituents that contribute to bioactivity [55].
• Gas chromatography–flame ionization detection (GC–FID) is often used in tandem with GC–MS for quantification purposes and for establishing fingerprint profiles [56].
• Fourier Transform Infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) spectroscopy may be used to confirm the presence of functional groups or detect adulterants [57].
• Chiral chromatography is employed to assess the stereoisomeric composition, which can influence bioactivity and sensory properties [58].

Establishing chemical markers and reference standards allows manufacturers and researchers to verify the identity and purity of EOs intended for food use.

2.4.5. Biological Activity Validation

Standardization also involves biological quality assessment, primarily when EOs are used for their antioxidant or antimicrobial properties in food preservation:

• Antioxidant activity is typically evaluated using assays such as DPPH, ABTS, ORAC, or FRAP, which must be consistently reproducible across batches [59].
• Antimicrobial efficacy can be assessed using disk diffusion, minimum inhibitory concentration (MIC), minimum bactericidal/fungicidal concentration (MBC/MFC), and time-kill kinetics. These bioassays are crucial for ensuring the effectiveness of EOs in various food matrices [60].

A standardized EO should have not only a consistent chemical profile but also a defined range of bioactivity under validated experimental conditions.

2.4.6. Detection of Adulteration and Contaminants

Due to their high economic value, EOs are frequently subject to adulteration. This can involve the following:

• Dilution with carrier oils or synthetic compounds;
• Addition of isolated natural constituents to mimic a full-spectrum EOs;
• Use of non-declared preservatives or solvents;
• Standardization protocols must include tests for authenticity and purity, using tools such as the following:
• Specific gravity, optical rotation, and refractive index;
• Stable isotope ratio analysis (SIRA) to distinguish synthetic from natural compounds;
• Residual solvent analysis via GC-MS or HS-SPME for detecting processing contaminants.

Additionally, testing for heavy metals, pesticide residues, and microbiological contamination is crucial for ensuring food-grade oils meet quality standards.

2.4.7. Role of Pharmacopeias and Standards Organizations

Several international bodies provide monographs and standards for EOs, including the following:

• European Pharmacopoeia (Ph. Eur.);
• United States Pharmacopoeia (USP);
• International Organization for Standardization (ISO);
• World Health Organisation (WHO) monographs.

These documents define specifications for botanical identity, extraction method, chromatographic fingerprint, and purity criteria, supporting both regulatory approval and industrial quality control.

For instance, ISO 4730 [61] specifies requirements for tea tree oil (Melaleuca alternifolia), including content of terpinen-4-ol, 1,8-cineole, and other constituents, along with limits on contamination and adulteration [62].

2.4.8. Stability and Shelf-Life Considerations

EOs can undergo oxidation, polymerization, or volatilization, leading to reduced efficacy and altered sensory properties [63]. Therefore, standardization also involves the following:

• Accelerated stability testing under light, heat, and humidity;
• Shelf-life determination under simulated storage conditions;
• Packaging optimization (e.g., amber glass, airtight seals).

Encapsulation technologies (e.g., nanoemulsions, liposomes) can enhance EO stability, but must be validated and standardized separately [64].

To ensure the safe and effective use of EOs in food preservation, a comprehensive standardization strategy is necessary. This includes both chemical and biological quality assessments, as well as authenticity verification and stability testing.

Table 2. Standardization and quality control techniques for essential oils used as food preservatives.

Category	Technique/Method	Purpose	Application
Chemical Characterization	GC-MS	Identify and quantify EO constituents	Establish EO fingerprint; detect major and minor compounds
	GC-FID	Quantify volatile components	Determine consistency in batches
	FTIR	Identify functional groups	Confirm composition and check for adulteration
	NMR	Structural elucidation of EO molecules	Authentication; detect processing artifacts
	Chiral Chromatography	Analyze enantiomeric composition	Detect changes in stereoisomeric forms affecting activity
Biological Validation	DPPH, ABTS, ORAC, FRAP assays	Assess antioxidant activity	Determine bioefficacy in oxidative food environments
	MIC/MBC/MFC, disk diffusion, time-kill assays	Assess antimicrobial potential	Predict preservation efficacy in food systems
Contaminant Detection	Residual solvent analysis (e.g., HS-SPME/GC-MS)	Detect solvents used during extraction	Ensure food-grade purity
	Heavy metal and pesticide screening	Identify chemical contaminants	Comply with safety standards for food ingredients
	Microbial load testing	Check for microbial contamination	Verify hygiene and shelf stability
Authenticity Testing	Specific gravity, refractive index, and optical rotation	Physical constants to verify identity	Detect dilution or substitution
	SIRA	Differentiate synthetic vs. natural compounds	Confirm natural origin
Stability Assessment	Encapsulation/nanoemulsion validation	Assess enhanced stability methods	Ensure consistency and bioavailability in final food products
	Accelerated stability tests (light, heat, humidity)	Evaluate degradation over time	Define shelf life and optimize packaging
Compliance Tools	ISO, USP, Ph. Eur., WHO Monographs	Standardized specifications for EO identity and purity	Support regulatory submissions and industrial quality assurance

ABTS: 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid; DPPH: 2,2-diphenyl-1-picrylhydrazyl; FRAP: Ferric Reducing Antioxidant Power; FTIR: Fourier Transform Infrared Spectroscopy; GC-FID: Gas Chromatography with Flame Ionization Detection; GC-MS: Gas chromatography-mass spectrometry; HS-SPME: Headspace Solid-Phase Microextraction; ISO: International Organization for Standardization; MBC: Minimum Bactericidal Concentration; MFC: Minimum Fungicidal Concentration; MIC: Minimum Inhibitory Concentration; NMR: Nuclear magnetic resonance; ORAC: Oxygen Radical Absorbance Capacity; Ph. Eur: European Pharmacopoeia; SIRA: Stable Isotope Ratio Analysis; USP: United States Pharmacopeia; WHO: World Health Organization.

summarizes the principal techniques used in the standardization and quality control of EOs, outlining their purpose and relevance for food applications.

3. Antioxidant Properties of Essential Oils

In oxidative food spoilage, the role of antioxidants is pivotal. Oxidation leads to rancidity, discoloration, off-flavors, and nutrient degradation, compromising shelf life and safety [65]. Traditionally, synthetic antioxidants such as BHT and BHA have been used to control these reactions; however, growing health and environmental concerns have led to an increased interest in natural alternatives. EOs, rich in phenolic and terpenoid compounds, present a promising option due to their inherent radical-scavenging capacity and lipid-peroxidation inhibition [66,67].

Several in vitro assays are routinely used to assess EO antioxidant capacity. Among them, the DPPH radical scavenging test and the ABTS decolorization assay are the most widely applied. For example, oregano and clove oils have demonstrated strong DPPH inhibition, while lemongrass and basil oils show high ABTS radical-quenching activity. These findings are often reported as IC₅₀ values or Trolox equivalent antioxidant capacities (TEAC), which allow comparative benchmarking [68,69,70,71,72].

However, in vitro efficacy does not always translate directly to complex food matrices [73]. Factors such as lipid content, water activity, and matrix interactions influence EO performance. Therefore, in situ validation, especially in lipid-rich foods like meat and dairy, is essential to confirm practical antioxidant effects under real-world conditions. Techniques such as lipid peroxidation assays (e.g., TBARS) are more relevant in these cases and help bridge the gap between laboratory data and food system application [74,75].

Significantly, antioxidant activity also contributes to EO stability, protecting volatile constituents from autoxidation and preserving their functional integrity during storage. Encapsulation strategies, such as nanoemulsions and biopolymer matrices, can further enhance stability while minimizing sensory impact.

Given the diverse physicochemical properties of food matrices, the efficacy of EOs as preservatives can vary significantly depending on whether the system is lipid-based or aqueous. The selection of appropriate analytical methods is therefore critical for accurately assessing EO performance in each context.

Table 3. Recommended analytical methods for evaluating essential oils in lipid-based vs. aqueous food matrices.

Food Matrix Type	Relevant Challenges	Recommended Methods	Notes
Lipid-based	Lipid oxidation	ORAC, FRAP, Lipid peroxidation assays	ORAC is suitable for peroxyl radicals commonly found in fats; FRAP is used for reducing power. Lipid peroxidation (e.g., TBARS) is a critical process.
	Sensory impact, EO stability	Encapsulation, Accelerated stability tests	Encapsulation helps control flavor and improve EO shelf life.
Aqueous-based	Microbial growth	Disk diffusion, MIC/MBC, ABTS	MIC/MBC are quantitative; ABTS works in both aqueous and lipid systems.
	Radical scavenging	DPPH, ABTS	DPPH is suited for hydrophilic systems; ABTS is more versatile.

ABTS: 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid; DPPH: 2,2-diphenyl-1-picrylhydrazyl; FRAP: Ferric Reducing Antioxidant Power; MBC: Minimum Bactericidal Concentration; MIC: Minimum Inhibitory Concentration; ORAC: Oxygen Radical Absorbance Capacity; TBARS: Thiobarbituric Acid Reactive Substances.

provides a comparative overview of the most suitable analytical techniques for lipid-rich and water-rich food systems, taking into account their specific preservation challenges.

3.1. Synergistic Potential and Stability Considerations

EOs often exhibit synergistic effects among constituents, such as between thymol and carvacrol or eugenol and citral. These interactions can amplify antioxidant effects while enabling lower effective doses, reducing sensory impact, and potential toxicity. Moreover, antioxidant activity contributes to EO stability itself, protecting active compounds from autoxidation. Novel delivery systems such as nanoemulsions and encapsulation technologies can further enhance stability and control release, ensuring prolonged functionality in complex food systems [76,77,78].

3.2. Sustainability Considerations: Toward Cleaner, Greener Food Products

The exploration of EOs as natural antioxidants aligns closely with the broader goals of sustainable food production [2]. The increasing awareness of the potential health risks and environmental burdens associated with synthetic antioxidants, such as BHA and BHT, has intensified the search for safer, plant-derived alternatives. EOs, particularly those sourced from renewable botanical materials and agricultural by-products, offer a promising solution by contributing to the development of clean-label food products with fewer chemical additives [79]. From a sustainability perspective, the incorporation of natural antioxidants can reduce the reliance on petrochemically derived substances and promote circular economy practices, mainly when EOs are extracted from previously underutilized plant materials or waste streams [80,81]. Moreover, their multifunctional nature—combining antioxidant, antimicrobial, and sensory-enhancing properties—may decrease the need for multiple synthetic additives, simplifying ingredient lists and minimizing ecological impact across the food supply chain [2].

The integration of antioxidant activity testing into EO research not only ensures functional efficacy but also supports regulatory approval and consumer acceptance. As such, evaluating and validating the antioxidant potential of EOs is a key step toward sustainable innovation in food preservation, reinforcing the transition toward greener technologies and healthier food systems.

4. Antimicrobial Activity of Essential Oils

Microbial spoilage remains a significant cause of food loss and safety concerns across the supply chain. EOs, with their rich composition of bioactive compounds, such as thymol, eugenol, citral, and carvacrol, demonstrate vigorous antimicrobial activity against a wide range of foodborne pathogens and spoilage organisms. Their use as natural antimicrobials aligns with both clean-label trends and sustainability goals by reducing reliance on synthetic preservatives and thermal treatments.

EOs exert antimicrobial effects through multiple mechanisms, including disruption of microbial membranes, leakage of cellular contents, inhibition of enzyme activity, and interference with quorum sensing. This multifaceted mode of action reduces the likelihood of resistance development, one of the key limitations of conventional antimicrobials [82,83,84,85,86,87].

4.1. Assessing Antimicrobial Potential

Several standard assays are used to evaluate the antimicrobial efficacy of EOs. Disk diffusion provides a quick, qualitative screening of inhibition zones, while minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC) tests offer more quantitative insights. Time-kill studies provide additional value by illustrating microbial reduction dynamics over time. While these tests are helpful for in vitro comparisons, they do not always reflect EO behavior in real food systems [88,89,90].

4.2. Performance in Food Matrices

EO effectiveness often diminishes in complex food environments due to interactions with fats, proteins, or carbohydrates that limit their bioavailability [91]. For instance, lipid-rich matrices like meat may require higher EO concentrations or encapsulated delivery systems to ensure efficacy without compromising sensory properties [92].

Despite these challenges, promising results have been reported:

• Oregano and thyme oils have been successfully used to reduce Listeria monocytogenes and Escherichia coli in ready-to-eat meat and dairy products [93,94].
• Clove and cinnamon oils have shown antifungal activity in bakery items and cereals [95,96].
• Lemongrass and tea tree oils have been applied in juices and fresh produce to suppress spoilage microorganisms [97,98,99].

The use of nanoemulsions, edible coatings, or active packaging films enhances EO dispersion and stability, supporting more consistent antimicrobial action while mitigating sensory concerns [100,101].

4.3. Multifunctionality and Safety Implications

EOs’ antimicrobial action is rarely isolated from their antioxidant and antigenotoxic functions. For example, clove essential oil not only inhibits microbial growth but also scavenges free radicals and reduces DNA damage from mycotoxins. This multifunctional profile supports the development of integrated food preservation strategies that simultaneously address safety, shelf life, and consumer health [102,103].

Additionally, by lowering microbial load, EOs may reduce the need for high-heat treatments or synthetic preservatives, contributing to both energy efficiency and nutrient retention in processed foods [104,105,106].

5. Antigenotoxic Potential of Essential Oils

Beyond their antimicrobial and antioxidant effects, EOs are increasingly recognized for their potential to protect cellular DNA from genotoxic damage [107]. This protective effect, referred to as antigenotoxicity, encompasses the ability of a substance to reduce, counteract, or prevent genetic damage caused by exposure to mutagens or genotoxins [108]. Genotoxicity involves any process that damages the genetic material within a cell, potentially leading to mutations, chromosomal alterations, or cancer [109]. In contrast, antigenotoxic agents may function through various mechanisms, including direct interaction with DNA-damaging agents (desmutagenesis), enhancement of DNA repair processes, modulation of detoxification pathways, or neutralization of reactive oxygen species that can induce DNA lesions. The antigenotoxic properties of EOs are therefore particularly relevant for food preservation, as they may help mitigate the harmful effects of foodborne contaminants such as mycotoxins, polycyclic aromatic hydrocarbons, and other processing-induced genotoxins. This antigenotoxic property is especially relevant in the context of food safety, as numerous food contaminants, such as mycotoxins, polycyclic aromatic hydrocarbons, heterocyclic amines, and nitrosamines, can cause oxidative stress and direct DNA lesions, leading to mutagenesis and carcinogenesis [107,110]. The ability of EOs to mitigate such damage introduces an additional functional dimension, reinforcing their value as multifunctional and sustainable food additives.

5.1. Foodborne Genotoxins and DNA Damage

Foods can be contaminated with a variety of genotoxic agents during production, processing, packaging, or storage [111]. Mycotoxins, such as aflatoxins produced by Aspergillus species, are among the most studied natural genotoxins and are common in grains, nuts, and dried fruits. PAHs, formed during grilling or smoking processes, and acrylamide, found in fried or baked goods, are additional examples of contaminants induced by processing [112,113]. These compounds are capable of forming DNA adducts, inducing strand breaks, or causing chromosomal abnormalities that may promote carcinogenesis over time.

The increasing detection of these agents in the food supply chain has raised concerns about their cumulative genotoxic risk. Consequently, food preservatives that not only prevent microbial spoilage but also actively protect against DNA damage are of significant interest in modern food science [107].

5.2. EOs and Genoprotection

EOs exhibit genoprotective effects, mainly due to their antioxidant and radical-scavenging activities, which can neutralize reactive oxygen and nitrogen species responsible for oxidative DNA damage [108,114,115]. Additionally, certain EO constituents may modulate detoxification enzymes, such as glutathione S-transferases, enhancing cellular defense mechanisms against mutagens [116,117].

Several studies have demonstrated that EOs or their main components—such as eugenol (from clove), carvacrol (from oregano), linalool (from lavender), and thymol (from thyme)—can significantly reduce DNA strand breaks or chromosomal aberrations induced by known genotoxins [118,119,120,121,122]. This protective effect can be observed in vitro using cell cultures and in vivo in model organisms, indicating a robust biological relevance.

5.3. Genotoxicity and Antigenotoxicity Assays

A variety of bioassays are used to evaluate both the genotoxic and antigenotoxic potential of substances, including EOs. The comet assay (single-cell gel electrophoresis) is one of the most sensitive methods for detecting DNA strand breaks at the single-cell level [123]. In this assay, cells treated with a test substance are lysed, electrophoresed, and stained to reveal “comet-like” tails, which reflect the extent of DNA migration due to strand breaks. EOs demonstrating shorter comet tails in toxin-exposed cells indicate a protective effect against DNA damage. The Ames test, which uses specific strains of Salmonella typhimurium to detect mutations, is another widely used method [124]. EOs are tested for their ability to reduce the mutagenicity of known carcinogens, indicating antigenotoxic potential through desmutagenic or bio-antimutagenic mechanisms. The micronucleus assay is employed to detect chromosomal damage or loss in dividing cells [124]. A reduction in micronuclei frequency after EO treatment in genotoxin-exposed cells provides further evidence of antigenotoxic effects. Together, these assays offer complementary insights into the safety profile and protective effects of EOs.

For instance, cinnamon essential oil combined with usnic acid in a nanoemulsion formulation led to a 50–60% reduction in micronucleus frequency in murine models exposed to genotoxic agents, demonstrating substantial antigenotoxic efficacy [125]. The antigenotoxic activity of oregano essential oil, confirmed in both comet and micronucleus assays, has also been validated in real food systems, including meat, dairy, and vegetable products, highlighting its practical relevance in food preservation strategies [126,127,128].

5.4. Implications for Food Safety and Human Health

The inclusion of EOs with antigenotoxic properties in food preservation strategies may offer additional health-related benefits; however, these should be viewed as emerging possibilities rather than confirmed outcomes. While in vitro and animal studies suggest that certain EOs can mitigate DNA damage caused by foodborne contaminants, such as mycotoxins or processing-induced mutagens [129,130,131], human data are currently lacking. Therefore, the potential of EOs to reduce the risk of mutation accumulation and associated chronic diseases (e.g., cancer) remains hypothetical at this stage. Continued investigation, particularly through human-relevant models and dietary exposure studies, is necessary to substantiate these effects. Nevertheless, the integration of antigenotoxicity screening into the development of natural preservatives supports a more preventive and safety-oriented approach to food technology.

6. Promising Essential Oils for Food Additives

The identification of EOs with strong and consistent bioactivity is crucial for the development of multifunctional preservation systems. This section highlights selected EOs that have demonstrated not only antimicrobial and antioxidant properties but also emerging antigenotoxic effects relevant to food safety and consumer health. Unlike traditional reviews, this section integrates these three biofunctional axes and links them to practical food applications, sustainability goals, and clean-label demands.

6.1. Selection Criteria and Evaluation Scope

The EOs were selected based on their compositional richness in bioactive compounds (e.g., phenols, aldehydes, terpenoids), in vitro and in situ efficacy data, and relevance across diverse food matrices. Selection also considered their distribution, established use, and regulatory acceptance in the food industry. Special attention was given to oils with validated performance in model food systems and those obtained from sustainable sources or food processing byproducts.

Key Essential Oils and Their Functional Potential

Oregano (Origanum vulgare): Rich in carvacrol and thymol, oregano EO exhibits strong antimicrobial activity against Listeria monocytogenes, Escherichia coli, and Staphylococcus aureus, as well as significant antioxidant and antigenotoxic potential [12,132]. It has been successfully applied in meat, dairy, and vegetable systems, particularly in encapsulated forms or edible coatings [133,134].

Thyme (Thymus vulgaris): Similar in composition to oregano, thyme EO is effective against Gram-positive and Gram-negative bacteria. Thymol and p-cymene contribute to both its antimicrobial and radical-scavenging actions [135,136]. Thyme essential oil also shows promise in reducing DNA damage in cell assays, suggesting antigenotoxic capability [137].

Clove (Syzygium aromaticum): Eugenol-dominant clove essential oil is among the most potent natural antioxidants, with documented antimicrobial action against spoilage fungi and bacteria [138]. It has reduced genotoxic markers in both comet and micronucleus assays, supporting its use in clean-label functional food applications [139].

Cinnamon (Cinnamomum zeylanicum or cassia): Cinnamaldehyde imparts cinnamon EO with antimicrobial properties effective against molds and pathogens such as Salmonella spp. and Aspergillus spp. [5]. It has also been shown to mitigate aflatoxin-induced DNA damage in vitro [140].

Rosemary (Rosmarinus officinalis): Known for its antioxidant molecules such as rosmarinic and carnosic acids, rosemary EO is widely used to stabilize lipids in meat and oil-based products [141]. While antimicrobial action is moderate, its antigenotoxic potential has been demonstrated in food models exposed to oxidative and genotoxic stress [142].

Tea Tree (Melaleuca alternifolia): Containing terpinen-4-ol and α-terpineol, tea tree EO offers broad-spectrum antimicrobial effects and moderate antioxidant activity. Preliminary antigenotoxic results are promising but require further study [143]. Applications include packaging films and salad dressings [144].

Lemongrass (Cymbopogon citratus): Citral-rich lemongrass EO shows notable antibacterial and antifungal effects, particularly in juices and fermented dairy [145]. Studies have also reported its radical-scavenging and DNA-protective properties [146].

Basil (Ocimum basilicum): Characterized by linalool and methyl chavicol, basil EO exhibits balanced antimicrobial and antioxidant activity. It is a promising candidate for fresh produce coatings and emulsified products [147]. Preliminary Ames test data suggest genoprotection [148].

Black pepper (Piper nigrum): Rich in piperine and other alkaloids, black pepper EO demonstrates antimicrobial activity against foodborne bacteria such as E. coli, Salmonella enterica, and Staphylococcus aureus. It also exhibits antioxidant potential, particularly in lipid-containing matrices, and is widely used as a flavoring agent in meat and snack products. Its broad culinary use and regulatory acceptance make it a relevant candidate for natural additive development [86,149].

Nutmeg (Myristica fragrans): Containing compounds such as myristicin, elemicin, and eugenol, nutmeg EO shows antimicrobial activity against Gram-positive bacteria and antioxidant effects in model food systems. It has also been tested in baked goods and dairy applications, supporting its integration into real-world formulations. Given its established use in the food industry, nutmeg EO aligns with clean-label expectations and multifunctional preservative strategies [150].

Table 4. Summary of selected essential oils for food preservation.

Essential Oil	Major Active Compounds	Antimicrobial Activity	Antioxidant Activity	Antigenotoxic Evidence	Common Food Applications	References
Oregano	Carvacrol, thymol	Strong (bacteria, fungi)	High	Comet, Micronucleus	Meat, dairy, and vegetables	[126,151,152,153]
Thyme	Thymol, p-cymene	Strong (Gram+ and Gram−)	High	Comet, Micronucleus	Juices, sauces, plant products	[154,155,156,157,158]
Clove	Eugenol	Strong	Very high	Ames, Comet	Bakery, meats, herbal drinks	[110,155,159,160,161]
Cinnamon	Cinnamaldehyde	Moderate to strong	High	Comet	Cereals, confectionery	[162,163,164,165,166]
Rosemary	Carnosic acid, rosmarinic acid	Moderate	Very high	Ames, Comet	Oils, processed meats	[165,167,168]
Tea Tree	Terpinen-4-ol, α-terpineol	Moderate	Moderate	Micronucleus, Comet	Packaging films, dressings	[169,170,171,172]
Lemongrass	Citral, limonene	Moderate	High	Comet	Juices, dairy drinks	[165,173,174,175]
Basil	Linalool, methyl chavicol	Moderate	Moderate	Ames	Fresh produce coating	[165,176,177]
Black pepper	Piperine, caryophyllene	Moderate to strong (especially against Gram-negative bacteria)	Moderate	Limited; in vitro data suggest DNA protection, but more studies are needed	Meat, sauces, snacks, spice blends	[178,179]
Nutmeg	Myristicin, elemicin, eugenol	Moderate (mainly Gram-positive bacteria and fungi)	Moderate	Preliminary in vitro evidence; further investigation required	Baked goods, dairy, desserts, beverages	[180,181]

Note: Terms such as “Strong,” “Moderate,” and “Very High” are qualitative descriptors based on reported efficacy in in vitro assays (e.g., MIC/MBC, DPPH, comet test) and application studies in food matrices. These reflect relative comparisons within the literature rather than fixed quantitative thresholds. Representative references are provided in the main text or reference list.

presents a comparative overview of selected EOs with their major bioactive compounds, key functional properties, and food application areas.

6.2. Strategic Implications for Food Preservation

Selecting the appropriate essential oil for a specific food system requires consideration of multiple factors, including organoleptic compatibility, matrix interactions, and regulatory status. The integration of EOs with proven antigenotoxic potential not only improves food preservation outcomes but also aligns with current trends toward cleaner labels and enhanced consumer health protection. Their multifunctional nature makes them excellent candidates for novel preservation systems, active packaging technologies, and functional food formulations that support both sustainability and safety.

7. Challenges and Limitations

Despite the growing interest and demonstrated efficacy of EOs in food preservation, several practical and scientific challenges must be addressed before their widespread adoption in commercial food systems. These challenges include issues related to sensory compatibility, chemical stability, standardization, regulatory frameworks, and safety.

7.1. Sensory Impact

One of the main limitations in the use of EOs in food preservation is their intense aroma and flavor, which can significantly alter the sensory profile of the final product [182]. EOs such as oregano, clove, and cinnamon are highly potent even at low concentrations, which may not be acceptable in foods with delicate or neutral flavor profiles. The strong organoleptic properties can compromise consumer acceptance, particularly when the EOs are not part of the traditional flavor system of the food product [132]. Encapsulation technologies, nanoemulsions, and active packaging are being explored to mask or control the release of volatile compounds, offering a promising route to minimize sensory disruption [183,184].

7.2. Stability During Processing and Storage

The volatile and thermolabile nature of many EO constituents makes them susceptible to degradation during food processing (e.g., heat, pH changes, light exposure) and storage [185]. For instance, compounds like carvacrol and citral can oxidize or evaporate, leading to a loss of bioactivity. This instability can reduce the effectiveness of EOs over time and pose challenges for ensuring consistent shelf-life protection. Emerging technologies, such as encapsulation in liposomes, cyclodextrins, or biopolymer matrices, are being investigated to enhance EO stability and facilitate controlled release in food environments [101].

7.3. Standardization of EO Composition

EOs are complex mixtures of dozens of bioactive molecules, and their composition can vary considerably depending on the plant species, chemotype, geographic origin, harvesting season, and extraction method. This chemical variability affects both efficacy and safety, making it difficult to reproduce results across studies or standardize industrial applications [48]. Greater investment in phytochemical profiling and quality control protocols is necessary, including the adoption of standardized markers and fingerprinting methods, such as GC-MS and HPLC, to ensure consistent composition and activity.

7.4. Regulatory Hurdles in Food Systems

Although many EOs and their major components are generally recognized as safe (GRAS) by authorities such as the United States FDA or authorized as flavoring agents in the EU, regulatory approval for use as food preservatives often requires additional toxicological and functional data. Different countries maintain divergent standards regarding acceptable levels, permitted food categories, and labeling requirements. Moreover, the dual use of EOs as both flavorings and preservatives complicates their classification under food law. More collaborative regulatory guidance and harmonization efforts are necessary to facilitate the safe and transparent use of EO applications in the global food market.

7.5. Potential Toxicity at High Doses

While EOs are natural, they are not inherently risk-free. Some constituents may exhibit cytotoxic, mutagenic, or allergenic effects when used at high concentrations. Several EOs contain aromatic compounds known to be skin or respiratory allergens, such as limonene, linalool, cinnamaldehyde, and eugenol. These substances can oxidize over time and form sensitizing compounds, which may lead to allergic contact dermatitis or respiratory irritation in individuals with sensitive skin [186,187]. Therefore, toxicological assessments, including NOAEL (No Observed Adverse Effect Level), chronic exposure studies, and interactions with food components, are essential to ensure safe use, especially in vulnerable populations such as children, older adults, or immunocompromised individuals.

7.6. Gaps in Antigenotoxicity Research

While the antigenotoxic potential of EOs is a promising area of investigation, current evidence is based mainly on in vitro assays and animal models, with limited validation in human dietary contexts. This restricts the direct extrapolation of results to real-world food applications. Moreover, antigenotoxic effects may be dose-dependent and influenced by the composition of food matrices, which can alter bioavailability and efficacy [188]. Further research is needed to explore these factors and to establish standardized protocols and human-relevant models for assessing the antigenotoxic safety and functionality of EOs in food systems.

8. Future Perspectives and Sustainability Outlook

EOs represent a compelling natural alternative to synthetic additives in food preservation, with potential benefits that extend beyond food safety into the realms of sustainability and the circular economy [189]. As the food industry seeks to align more closely with environmental and health-conscious goals, the strategic integration of EOs into sustainable processing systems offers a promising path forward. This section discusses key opportunities for future development, including technological innovation, resource utilization, and alignment with global sustainability frameworks.

8.1. Integration into Green Food Processing

One of the most significant opportunities lies in integrating EOs into green food processing technologies, which prioritize energy efficiency, minimal use of synthetic chemicals, and a low environmental impact [190]. EOs can be incorporated into eco-friendly preservation methods, such as cold plasma, pulsed electric fields, or high-pressure processing, serving as natural antimicrobials or antioxidants in synergistic combinations [107]. Moreover, embedding EOs in biodegradable films, edible coatings, and smart packaging systems enables controlled release and targeted action, reducing waste and enhancing shelf life without compromising product integrity [191]. These innovations can contribute to cleaner production chains and a reduced carbon footprint.

8.2. Valorization of Agricultural Byproducts

The extraction of EOs from agricultural residues, such as citrus peels, herb trimmings, or discarded leaves that retain sufficient volatile compounds, supports the principles of the circular economy [190,191]. While not all agricultural byproducts are suitable for EO extraction due to the loss of volatile constituents during processing, some aromatic residues can still be effectively valorized. In other cases, these byproducts may be more appropriate for producing extracts rich in non-volatile bioactive substances (e.g., polyphenols, flavonoids) rather than true essential oils. For instance, orange peels—commonly discarded in juice production—are rich in limonene and can be processed into EOs with antimicrobial and aromatic properties. Depending on the extraction method and peel moisture content, approximately 3–10 kg of limonene-rich essential oil can be obtained from one ton of fresh citrus peels [26]. This high-yield recovery from agro-industrial waste highlights the potential of valorization strategies to significantly reduce food system waste and contribute to circular economy goals. This selective yet high-yield recovery from specific agro-industrial waste streams underscores the potential of targeted valorization strategies to significantly reduce food system waste and contribute to achieving circular economy goals. This approach enhances the sustainability of food supply chains by generating economic value from biomass and reducing reliance on land-intensive primary crops.

8.3. Contribution to Sustainable Development Goals (SDGs)

The application of EOs in food preservation aligns with several United Nations SDGs, specifically, the following:

• SDG 3 (Good Health and Well-Being): EOs contribute to safer food by reducing microbial contamination and potentially mitigating DNA damage from foodborne genotoxins.
• SDG 12 (Responsible Consumption and Production): Using natural, renewable preservatives promotes sustainable food systems and reduces chemical dependency.
• SDG 13 (Climate Action): Adoption of green extraction and processing methods using EOs can lower energy use and greenhouse gas emissions.
• SDG 9 (Industry, Innovation and Infrastructure): Advances in EO encapsulation, packaging, and formulation stimulate innovation across the food and biotech sectors.

By fostering safer, more natural, and environmentally friendly food preservation strategies, EOs hold the potential to be key enablers of the food industry’s sustainability transformation [187,192].

8.4. Research Needs and Innovation Gaps

Despite promising results, the transition to wide-scale application of EOs in food preservation demands further research and standardization. Key gaps include the following:

• Dose Optimization: There is a need for detailed studies to determine minimum effective concentrations in complex food matrices, while ensuring sensory acceptability and safety.
• Consumer Acceptance: While natural preservatives are generally welcomed, consumer perception of intense EO flavors or odors can be a barrier. Sensory studies and transparent communication are crucial for building trust.
• Life Cycle Assessment (LCA): Comprehensive LCA studies comparing EO-based systems to synthetic preservatives are still limited. These assessments are crucial for quantifying environmental benefits and informing decision-making.
• Synergistic Formulations: Research into EO combinations and their interactions with other natural agents (e.g., organic acids, bacteriocins) can enhance efficacy and reduce the required dosages.

Continued collaboration between academia, industry, and regulatory bodies is essential to translate laboratory findings into scalable, consumer-friendly, and environmentally responsible applications.

8.5. Addressing Inconsistencies and Standardization Needs

Although numerous studies highlight the potential of essential oils in food preservation, outcomes can vary significantly due to differences in oil composition, extraction methods, test organisms, food matrices, and experimental conditions. Some applications have failed to demonstrate consistent efficacy, particularly under real food system constraints. This variability highlights the urgent need for harmonized testing protocols, including standardized EO characterization, bioassays, and validation in model and commercial food products. Establishing such guidelines would enhance reproducibility, facilitate regulatory approval, and accelerate responsible adoption of EO-based preservatives.

9. Conclusions

EOs have emerged as versatile and promising natural agents for food preservation, offering antimicrobial, antioxidant, and antigenotoxic properties that support both food safety and consumer health. Their broad-spectrum bioactivity, coupled with their natural origin, positions EOs as viable alternatives to synthetic preservatives in the context of an increasing demand for clean-label and eco-conscious food products.

This review has highlighted the complex chemical composition of EOs, their mechanisms of action, and the array of assays used to evaluate their efficacy. It has also examined their role in reducing microbial contamination, oxidative spoilage, and genotoxic risk from foodborne contaminants—each of which is central to long-term food safety. Importantly, the integration of EOs into green food processing technologies and their potential extraction from agricultural byproducts offers a clear link to sustainability goals and circular economy principles. This review stands out by integrating the emerging concept of antigenotoxicity into the narrative of food preservation.

Nevertheless, several challenges remain, including sensory impact, stability issues, compositional variability, regulatory hurdles, and potential toxicity at high doses. Addressing these challenges will require coordinated research efforts focused on formulation science, dose optimization, and consumer perception. Additionally, life cycle assessments and real-world validation in various food matrices are essential for a more complete understanding of EO-based preservation systems.

Looking forward, EOs have the potential to reshape food preservation practices by aligning scientific innovation with public health and environmental priorities. Future efforts must focus on bridging regulatory gaps, enhancing consumer education, and developing scalable encapsulation technologies that are effective and sustainable. With further research and strategic implementation, they can make meaningful contributions to the development of safer, more natural, and sustainable food systems.