Abstract Maximizing the extraction efficiency of bioactive compounds from botanical matrices must inherently align with environmental sustainability. This study evaluates the ecological profile of a previously established, optimized ultrasound-assisted extraction process for Artocarpus altilis leaves, designated as the A. altilis extraction process (AAEP) using a gate-to-gate Life Cycle Assessment (LCA). Executed via openLCA software and the ILCD 2011 Midpoint+ method, the assessment quantified 15 impact categories. Results indicated that while grid electricity and the ethanol supply chain drove climate change (2.59E-01 kg CO₂ eq) and freshwater ecotoxicity (9.92E-02 CTUe), the overall environmental burdens remained remarkably low. Normalized and weighted analyses pinpointed human toxicity (cancer effects), photochemical ozone formation, and water resource depletion as the primary hotspots; however, their absolute magnitudes were infinitesimal (below the 10^-4 threshold). By completely eliminating hazardous organic solvents, the AAEP strictly adheres to Green Chemistry principles and Sustainable Development Goals (SDGs 3 and 12), establishing a highly benign, eco-friendly framework for the greener valorization of natural products.

Keywords: Artocarpus altilis, Environmental impacts, Green chemistry, Life cycle assessment

Citation:  Duc, C.K.T. and Tran Linh, T.C. 2026. Life cycle assessment of the optimized extraction process from Artocarpus altilis leaves. Natural and Life Sciences Communications. 25(4): e2026078.

Graphical Abstract:

INTRODUCTION

Artocarpus altilis, a plant species belonging to the Moraceae family and widely distributed across tropical regions, has long been recognized as a prolific source of bioactive compounds, particularly polyphenols (including flavonoids) (Lan et al., 2013; Soifoini et al., 2018). These compounds are well-known for their diverse biological activities, including potent antioxidant, anti-inflammatory, and antidiabetic properties (Musa et al., 2025; Auranwiwat et al., 2026; Ayub et al., 2026). To maximize the pharmacological value of A. altilis leaves, our previous study successfully developed an ultrasound-assisted extraction (UAE) process coupled with Response Surface Methodology (RSM) employing a Box-Behnken design (Lan and Linh, 2022). The results demonstrated that under the optimized operating conditions, the recovery yields of total polyphenol content (TPC) and total flavonoid content (TFC) reached their maxima at 227.38 ± 1.27 mg GAE/g extract and 113.36 ± 2.07 mg QE/g extract, respectively (Lan and Linh, 2022). These experimental values were highly consistent with the theoretical predictive models, and the resulting extracts also exhibited remarkable in vitro antioxidant activity (Lan and Linh, 2022).

Although the aforementioned process maximized the yield of TPC and TFC from A. altilis leaves, focusing solely on recovery efficiency is insufficient to affirm the technological sustainability through the lens of Green Chemistry (Kurul et al., 2025). In practice, laboratory-scale extraction processes for medicinal plants frequently consume substantial amounts of energy and organic solvents (Phongthai et al., 2025; Yulianti et al., 2026). These factors introduce potential ecological burdens that are often underestimated or overlooked in conventional optimization studies. Therefore, conducting follow-up analyses on the environmental profile of the extraction process is an urgent requirement, aligning with the United Nations Sustainable Development Goals (SDGs), notably SDG 3 (Good Health and Well-being), and SDG 12 (Responsible Consumption and Production) (Cordella et al., 2023). Strikingly, to date, there have been no scientific reports comprehensively evaluating the environmental impacts of the extraction processes for bioactive compounds from A. altilis leaves.

Stemming from the aforementioned research gap, this study was conducted to apply the Life Cycle Assessment (LCA) framework in accordance with the international ISO 14040/14044 standards to analyze the life cycle of the established optimal extraction process. The core objective of this paper is to quantify the environmental impacts and identify the “hotspots” concerning resource consumption and emissions throughout the entire process. Based on this foundation, the study will provide insightful discussions and propose feasible improvement scenarios. This approach aims to develop a greener technological process that ensures environmental benignity while maintaining high recovery efficiency of pharmacologically valuable compounds (such as natural antioxidants and active pharmaceutical ingredients) (Duc et al., 2025). This comprehensive approach not only provides guidance for future scale-up operations but also reinforces the profound interdisciplinary synergy between Natural Product Chemistry and Green Chemistry.

MATERIALS AND METHODS

Optimized extraction process

The present study utilizes the optimal parameters established in our previously published work on the extraction of A. altilis leaves (Lan and Linh, 2022), as the baseline model for the LCA. Briefly, the extraction process was conducted on dried A. altilis leaves using a 40 kHz heated ultrasonic bath (Derui DR-MH30, China). The specific operational conditions were set at an ethanol concentration of 69%, a solid-to-solvent ratio of 1/30 (w/v), a temperature of 69°C, and a sonication time of 15.98 min (Lan and Linh, 2022), serves as the foundational inventory for evaluating the environmental impacts in this study.

Life cycle assessment

Goal and scope definition
This LCA aims to evaluate the potential environmental impacts associated with the laboratory-scale optimized extraction process of A. altilis leaves, previously developed by our research group (Lan and Linh, 2022). The functional unit (FU) was defined as 1 g of dried raw material (dried A. altilis leaves). While a product-based FU is often used to evaluate end-product efficiency, this input-based FU was specifically selected to maintain consistency with our previous LCA studies (Duc et al., 2025; Duc et al., 2026; Thanh et al., 2026; Thien Duc and Linh, 2026; Linh et al., 2026). Furthermore, it provides a clearer baseline for objectively assessing the environmental impacts of the early processing steps (e.g., raw material grinding and preparation) before the actual extraction phase. A “gate-to-gate” approach was adopted for the system boundary (Figure 1), which strictly encompasses the operations from the intake of dried leaves into the laboratory through to the final extract recovery (Duc et al., 2025; Duc et al., 2026; Linh et al., 2026; Thanh et al., 2026). This explicitly includes milling, the optimized extraction process, gravity filtration, and vacuum rotary evaporation. Upstream agricultural processes (plant cultivation) and downstream end-of-life waste management were excluded from the scope.

Figure 1. LCA framework for the optimized extraction process of A. altilis leaves.

Life cycle inventory (LCI)

Primary inventory data (input), encompassing the consumption of chemicals (water and ethanol) along with the electrical energy required by the processing equipment (milling machine, ultrasonic bath, and vacuum rotary evaporator) (Figure 1), were directly acquired from our preceding optimization trials (Lan and Linh, 2022). Secondary data associated with all these inputs were sourced from the ELCD 3.2 background database (Duc et al., 2026; Thanh et al., 2026).

Life cycle impact assessment (LCIA)

The LCIA (output) (Figure 1) was conducted using the openLCA 2.4.1 software, employing the ILCD 2011 Midpoint+ method (Duc et al., 2026). The impact categories include: acidification; climate change; freshwater ecotoxicity; freshwater eutrophication; human toxicity, cancer effects; human toxicity, non-cancer effects; ionizing radiation HH; land use; marine eutrophication; mineral, fossil & ren resource depletion; ozone depletion; particulate matter; photochemical ozone formation; terrestrial eutrophication; water resource depletion. Following the characterization analysis, the impact values were normalized to convert the results into a unified, comparable scale. This normalization phase was conducted using the EC-JRC Global, equal weighting, integrated within the openLCA 2.4.1.

Interpretation  

In the final phase of the LCA framework, an interpretation was conducted to systematically identify, quantify, and evaluate the most significant environmental hotspots within the optimized extraction process. The normalized LCIA results and inventory data were cross-analyzed to determine the proportional contributions of each operational input, specifically, electrical energy consumption and chemical solvents, to the selected impact categories. Ultimately, based on these analytical findings, the study proposes actionable recommendations for process improvements and sustainable scale-up strategies aligned with Green Chemistry principles (Martinengo et al., 2024; Kurul et al., 2025).

Software and data visualization

The LCA modeling and impact analysis were executed utilizing openLCA 2.4.1 software (GreenDelta, Berlin, Germany), while all flowchart diagrams and structural system boundaries were visualized using Microsoft PowerPoint 2016 (Microsoft Corporation, WA, USA).

RESULTS AND DISCUSSION

Rationale and framework for environmental assessment

While optimizing operational parameters is essential for maximizing the recovery of bioactive compounds from botanical sources (Linh et al., 2025), modern extraction processes must transcend mere efficiency criteria. In our previous work, an optimized protocol for extracting active compounds from A. altilis leaves was successfully established (Lan and Linh, 2022). However, to realize a truly sustainable technological process, one that safeguards human health while mitigating ecological burdens, the implementation of a Life Cycle Assessment (LCA) is imperative (Duc et al., 2025). Notably, our established procedure employs ultrasound-assisted extraction (UAE) coupled with an ethanol/water solvent system; both are highly regarded for their environmental benignity (Lajoie et al., 2022; Shen et al., 2023), aligning closely with the core principles of Green Chemistry (Martinengo et al., 2024; Kurul et al., 2025).

This LCA framework has been successfully implemented by our research group in a series of recent publications to analyze the environmental profiles of extraction processes from diverse biomass sources, including medicinal plants, agricultural by-products, rhizomes, and flowers, as well as the fabrication of biomedical materials. Notable examples encompass comprehensive environmental sustainability assessments of the optimized recovery of polyphenols and flavonoids from Ehretia asperula leaves (Duc et al., 2025); cellulose extraction from lotus seedpod (Nelumbo nucifera) waste (Duc et al., 2026); and flavonoid-enriched extractions from Curcuma zedoaria rhizomes (Thanh et al., 2026) and Mansoa alliacea flowers (Linh et al., 2026). Particularly, this methodology has also been effectively deployed to evaluate the green preparation of silk fibroin-based nanoparticles (Giang et al., 2025; Pham et al., 2025).

To the best of our knowledge, no prior scientific literature has applied LCA to evaluate the environmental impacts of extraction processes targeting A. altilis leaves. This scarcity of species-specific data makes direct comparisons challenging. To overcome this barrier, we benchmarked our findings against previously published LCA values from other botanical matrices processed under comparable operational conditions (Duc et al., 2025). Specifically, because the reference study also utilized UAE, this selection ensures a more equitable evaluation. Comparing UAE directly with another UAE process eliminates the severe biases in baseline electricity consumption associated with fundamentally different equipment architectures, such as Soxhlet or microwave-assisted extraction. This comparative approach furnishes an objective baseline, enabling a precise quantification of the “greenness” and overall sustainability of the A. altilis extraction protocol proposed herein. The following flowchart outlines the extraction protocol established in our previous research (Lan and Linh, 2022), detailing the specific electricity and chemical consumption (Figure 2). These parameters constitute the fundamental inputs for the LCI, facilitating a rigorous quantification of the potential environmental footprints and human health risks (LCIA) associated with the A. altilis extraction process (AAEP).

Figure 2. Flowchart of the A. altilis extraction process (AAEP) outlining specific electricity and chemical consumption per functional unit.

Environmental impacts

The life cycle impact assessment (LCIA) results of the AAEP are detailed in Table 1. An analysis of the primary environmental hotspots (high-impact categories) reveals that Climate change (2) emerges as a notable ecological burden, recording a value of 2.59E-01 kg CO₂ eq. This is closely followed by the indicators for Freshwater ecotoxicity (3) (9.92E-02 CTUe) and Ionizing radiation HH (7) (2.01E-02 kBq U235 eq). The root causes of these hotspots do not stem from the chemical system itself, but primarily arise from two indirect factors. First, the process currently relies heavily on the national power grid to operate energy-intensive equipment such as the sample mill, the UAE system, and notably, the prolonged rotary evaporation process during the solvent recovery stage (Figure 2). Given that the background grid still operates predominantly on fossil fuel combustion, this electricity consumption translates directly into substantial emissions of greenhouse gases (CO₂, CH₄) and acidifying oxides (Majumdar and Gajghate, 2011; Jeon, 2022). Second, the elevated toxicity and radiation indicators are an inevitable consequence of the underlying energy production infrastructure, coupled with the agricultural cultivation activities within the ethanol solvent supply chain (Osman et al., 2024). However, these hotspots must be viewed in a relative context. When benchmarked against our research group's previously published optimal UAE process for E. asperula leaves (Duc et al., 2025), the core impact indicators of the AAEP, particularly climate change potential and freshwater ecotoxicity, are noticeably lower. This comparison provides objective evidence that the AAEP generally maintains a highly favorable environmental profile, does not impose severe ecological burdens.

An interesting highlight in the environmental profile of the AAEP is the moderate level of impact in categories related to ecosystems and natural resources. This includes Land use (8) at 2.37E-03 kg C deficit, Water resource depletion (15) reaching 9.95E-03 m³ water eq, and Eutrophication indicators, which recorded 1.83E-03 molc N eq for the terrestrial environment (14) and 1.50E-04 kg N eq for the marine environment (9) (Table 1). This moderate ecological burden is intrinsically linked to the upstream supply chain of the chosen extraction solvent, ethanol (Figure 2). The production life cycle of ethanol typically encompasses resource-intensive phases, including substantial water consumption and processes involving nitrogen and phosphorus emissions (Bhatia et al., 2012; Falano et al., 2014; Osman et al., 2024). These background production phases are the primary contributors to soil and freshwater eutrophication, as reflected in indicator (4) at 4.17E-06 kg P eq. Furthermore, the inherent volatility of ethanol during the extraction and solvent recovery stages directly drives Photochemical ozone formation (13) (1.14E-02 kg NMVOC eq) within the operating area. Acknowledging these environmental trade-offs underscores that no single solvent system is absolutely perfect; rather, it requires making the most suitable choice for a specific botanical matrix. Despite these inherent background burdens, the AAEP maintains a highly competitive environmental profile when benchmarked against a comparable reference process. Specifically, the terrestrial and marine eutrophication indicators of the AAEP are 2.2 times and 2.4 times lower, respectively, than those of the reference process (which reached 4.1E-03 mol N eq and 3.6E-04 kg N eq, respectively) (Duc et al., 2025). Most notably, the freshwater eutrophication burden of the AAEP (only 4.17E-06 kg P eq) proves highly superior, being nearly 20 times lower than the 7.9E-05 kg P eq recorded for the reference process (Duc et al., 2025).

Disregarding energy-related limitations, the greatest success of the AAEP process is reflected in the dramatic reduction of toxicity indicators. The complete elimination of traditional toxic organic solvents (such as chloroform or n-hexane) has brought Human toxicity (5) for cancer effects and Human toxicity (6) for non-cancer effects down to negligible levels (8.94E-09 and 9.49E-09 CTUh, respectively). Simultaneously, the process demonstrates excellent preservation of the ozone layer, with Ozone depletion (11) at 1.46E-08 kg CFC-11 eq, and Mineral, fossil & ren resource depletion (10) at 2.57E-08 kg Sb eq. These extremely modest figures not only confirm strict adherence to the principles of Green Chemistry (use of safe solvents) (Martinengo et al., 2024; Kurul et al., 2025) but also ensure absolute feasibility and safety when applying this process to produce pharmaceuticals or functional foods for direct human consumption.

In summary, the LCIA results confirm that the AAEP is a highly sustainable extraction strategy. By minimizing the footprint across 15 categories, notably human toxicity and resource preservation, this process strictly aligns with the Principles 5 and 6 of Green Chemistry (Martinengo et al., 2024; Kurul et al., 2025). Furthermore, the AAEP directly supports SDG 3 and SDG 12 (Giannetti et al., 2020; Cordella et al., 2023), by eliminating hazardous waste, offering a robust, eco-friendly model for producing safe bioactive compounds within planetary boundaries.

Table 1. LCIA characterization results of the AAEP.

Note: molc H⁺ eq: Moles of charge of hydrogen equivalent; kg CO₂ eq: Kilograms of carbon dioxide equivalent; CTUe: Comparative Toxic Unit for ecosystems; kg P eq: Kilograms of phosphorus equivalent; CTUh: Comparative Toxic Unit for humans; kBq U235 eq: Kilobecquerels of uranium-235 equivalent; kg C deficit: Kilograms of carbon deficit; kg N eq: Kilograms of nitrogen equivalent; kg Sb eq: Kilograms of antimony (Stibium) equivalent; kg CFC-11 eq: Kilograms of trichlorofluoromethane (CFC-11) equivalent; kg PM_2.5 eq: Kilograms of particulate matter (diameter ≤ 2.5 µm) equivalent; kg NMVOC eq: Kilograms of non-methane volatile organic compounds equivalent; molc N eq: Moles of charge of nitrogen equivalent; m³ water eq: Cubic meters of water equivalent.

Normalization and weighting analysis of environmental impacts

To further evaluate the relative significance of the environmental burdens identified in the LCIA stage (Table 1), a normalization and weighting analysis was performed (Table 2). This step is essential to transform characterized results with different units into a dimensionless format (Matuštík et al., 2024), enabling a direct comparison across all 15 impact categories. In this study, the ILCD 2011 Midpoint+ method was utilized, integrated within the openLCA 2.4.1 software platform. The normalization and weighting factors were based on the EC-JRC Global, equal weighting set, which provides a balanced perspective on global environmental priorities (Linh et al., 2026). By aggregating these results into a single score (measured in Pt), the primary environmental “hotspots” of the AAEP can be objectively identified, offering a clear baseline for assessing its overall sustainability and guiding future process optimizations.

The normalized results presented in Table 2 facilitate a comprehensive cross-category comparison by expressing each environmental impact relative to a standardized reference load. Based on these values, Human toxicity, cancer effects (5) was identified as the most dominant impact category, yielding a value of 7.21E-04. This magnitude is substantially higher than all other assessed categories, signifying that emissions of substances with carcinogenic potential represent the primary environmental bottleneck within the AAEP. However, from a toxicological and life-cycle perspective, this pronounced impact is not derived from direct emissions during the AAEP, but is rather an indirect consequence of upstream resource consumption, predominantly electricity and chemical manufacturing (Figure 2). The second most significant contribution was observed for Photochemical ozone formation (13), with a value of 2.53E-04. This suggests a notable influence of atmospheric precursor emissions, such as volatile organic compounds (VOCs) or nitrogen oxides (NO_x) (David and Niculescu, 2021; An et al., 2024), which are critical factors in urban air quality modeling. This was followed by Water resource depletion (15), which reached 1.44E-04, indicating that the cumulative water consumption during the production stages constitutes a relevant environmental pressure that warrants further optimization. Furthermore, moderate normalized impacts were recorded for Ionizing radiation HH (7) (8.34E-05) and Human toxicity, non-cancer effects (6) (6.12E-05). While Climate change (2) remains a global priority, it presented a relatively lower but still statistically notable contribution of 3.67E-05 in this specific system. In stark contrast, several impact categories exhibited negligible normalized values, most notably Freshwater eutrophication (4) (6.38E-07) and Land use (8) (4.56E-10). These infinitesimal values confirm that such environmental pressures play a minor role in the overall ecological profile of the AAEP, allowing researchers to focus mitigation strategies on the higher-ranking toxicological and resource-related categories.

Upon applying weighting factors, the environmental impacts are expressed as single scores (Pt), providing a more robust evaluation of each category's contribution to the overall environmental burden of the system. Consistent with the normalized findings, all weighted values remained extremely low (Table 2). Among the assessed impact categories, Human toxicity, cancer effects (5) continued to be the most dominant factor, yielding a weighted value of 4.81E-05 Pt, the highest recorded in the entire profile. This was followed by Photochemical ozone formation (13) at 1.68E-05 Pt and Water resource depletion (15) at 9.63E-06 Pt. These categories represent relatively higher contributions compared to the rest of the profile, although their absolute magnitudes remain within a low environmental impact threshold. Furthermore, several categories showed notable but lower weighted contributions, including Ionizing radiation HH (7) at 5.56E-06 Pt and Human toxicity, non-cancer effects (6) at 4.08E-06 Pt. Categories such as Freshwater ecotoxicity (3) (1.77E-06 Pt), Acidification (1) (1.45E-06 Pt), and Terrestrial eutrophication (14) (7.42E-07 Pt) exhibited relatively minor contributions to the total single score. The remaining categories showed negligible weighted values; for instance, Marine eutrophication (9) reached 3.28E-07 Pt, Ozone depletion (11) reached 7.97E-08 Pt, and Freshwater eutrophication (4) stood at 4.25E-08 Pt. Most notably, Land use (8) recorded an infinitesimal value of 3.04E-11 Pt, indicating that this impact is virtually insignificant within the overall environmental profile of the system. In summary, the weighted and single score results indicate that the primary environmental impacts of the assessed system are linked to human toxicity, atmospheric pollution processes, and water resource consumption. However, the fact that all obtained values are remarkably low reflects that the evaluated production process exerts a relatively minimal environmental footprint within the defined research scope.

Table 2. Normalized and weighted results of the AAEP.

Overall, the comprehensive LCA demonstrates that the AAEP possesses a remarkably benign environmental profile. Both the normalized and weighted single-score analyses consistently pinpoint human toxicity (cancer effects), photochemical ozone formation, and water resource depletion as the system's primary environmental hotspots. However, the extraordinarily low absolute magnitudes of these impacts, remaining below the 10^-4 threshold even for the most dominant categories, substantiate the minimal ecological footprint of the assessed system. These findings validate the environmental viability of the proposed extraction protocol, confirming that the optimized process from A. altilis leaves (Lan and Linh, 2022) aligns seamlessly with the core principles of Green Chemistry as well as Sustainable Development Goals (Giannetti et al., 2020; Cordella et al., 2023; Martinengo et al., 2024; Kurul et al., 2025). By effectively mitigating toxicological risks and resource consumption, the AAEP establishes a sustainably sound framework for advancing its subsequent pharmacological applications and guiding its potential scale-up. Nonetheless, translating this lab-scale framework into industrial reality must carefully navigate the inherent “scale-up gap,” where pilot-scale validations and efficient solvent recovery engineering are essential.

Limitations and future perspectives

While the current AAEP exhibits a highly benign environmental profile, several limitations present opportunities for future optimization. First, although the established protocol is ecologically sound, future investigations could explore the substitution of current solvents with emerging green extraction media, such as natural deep eutectic solvents (NADES) (Stanisz et al., 2024), room-temperature ionic liquids (RTILs) (Azarhoosh and Pirsa, 2026), or supercritical carbon dioxide (Čižmek et al., 2021). The integration of these alternative solvents could further drive down the already low human toxicity and ecotoxicity indicators. Second, the residual environmental burdens, particularly concerning photochemical ozone formation and water resource depletion, could be substantially alleviated by transitioning the manufacturing energy supply from conventional fossil-fuel-reliant grids to renewable energy sources, such as solar photovoltaics or wind power (Dupré la Tour, 2023). Furthermore, the system boundary of this study was conceptually restricted to a gate-to-gate approach, focusing exclusively on the extraction and processing phases. To provide a more holistic environmental panorama, subsequent life cycle assessments should broaden this scope to a cradle-to-gate or cradle-to-grave framework (Zhou et al., 2022). Finally, another limitation of this study is the reliance on the European-centric ELCD 3.2 database due to the lack of localized LCI datasets. While this geographical discrepancy may underestimate absolute environmental impacts (e.g., due to differences in local electricity grid mixes), applying this uniform baseline does not affect the relative comparison or the final conclusions regarding the optimized extraction process.

CONCLUSION

This study successfully quantified the environmental profile of the AAEP using a comprehensive gate-to-gate LCA. The quantitative findings definitively substantiate that the AAEP a remarkably benign ecological footprint. Although normalized and weighted single-score analyses pinpointed human toxicity (cancer effects), photochemical ozone formation, and water resource depletion as the system's primary environmental hotspots, their absolute magnitudes remained exceptionally low (below 10^-4 for normalized values and 10^-5 Pt for weighted scores). By eliminating traditional toxic organic solvents, this extraction protocol effectively mitigates severe ecological burdens, aligning seamlessly with the core principles of Green Chemistry and the UN Sustainable Development Goals (SDGs 3 and 12). Ultimately, the AAEP establishes a robust, sustainable foundation for the valorization of A. altilis leaves into pharmacologically valuable extracts. To further drive down the residual environmental impacts, future research and industrial scale-up strategies should focus on substituting conventional solvents with emerging green media (such as natural deep eutectic solvents or supercritical carbon dioxide), transitioning to renewable energy grids, and expanding the assessment boundary to a holistic cradle-to-gate framework.

AUTHOR CONTRIBUTIONS

Chong Kim Thien Duc: Conceptualization (Equal), Methodology (Equal), Investigation (Equal), Data Curation (Equal), Validation (Equal), Writing – Original Draft (Equal), Writing – Review & Editing (Equal); Tran Chi Linh: Conceptualization (Equal), Methodology (Equal), Investigation (Equal), Data Curation (Equal), Validation (Equal), Project Administration (Lead), Resources (Lead), Writing – Original Draft (Equal), Writing – Review & Editing (Equal). All authors have read and agreed to the published version of the manuscript.

CONFLICT OF INTEREST

The authors declare that they have no conflicts of interest.

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