Supercritical Fluid Extraction Research Articles

A short review of supercritical fluid extraction of plant extracts

A short review of supercritical fluid extraction of plant extracts

Unveiling the mechanisms for the development of rosehip-based dermatological products: an updated review

Rosa spp., commonly known as rosehips, are wild plants that have traditionally been employed as herbal remedies for the treatment of a wide range of disorders. Rosehip is a storehouse of vitamins, including A, B complex, C, and E. Among phytonutrients, vitamin C is found in the highest amount. As rosehips contain significant levels of vitamin C, they are perfect candidates for the development of skincare formulations that can be effectively used in the treatment of different skin disorders (i.e., scarring, anti-aging, hyperpigmentation, wrinkles, melasma, and atopic dermatitis). This research focuses on the vitamin C content of several Rosa sp. by their botanical and geographic origins, which according to research studies are in the following order: R. rugosa > R. montana > R. canina > R. dumalis, with lower levels in R. villosa and R. arvensis, respectively. Among rosehip species, R. canina is the most extensively studied species which also displays significant amounts of bioactive compounds, but also antioxidant, and antimicrobial activities (e.g., against Propionibacterium acnes, Staphylococcus aureus, S, epidermis, and S. haemolyticus). The investigation also highlights the use of rosehip extracts and oils to minimise the harmful effects of acne, which primarily affects teenagers in terms of their physical appearance (e.g., scarring, hyperpigmentation, imperfections), as well as their moral character (e.g., low self-confidence, bullying). Additionally, for higher vitamin C content from various rosehip species, the traditional (i.e., infusion, maceration, Soxhlet extraction) and contemporary extraction methods (i.e., supercritical fluid extraction, microwave-assisted, ultrasonic-assisted, and enzyme-assisted extractions) are highlighted, finally choosing the best extraction method for increased bioactive compounds, with emphasis on vitamin C content. Consequently, the current research focuses on assessing the potential of rosehip extracts as medicinal agents against various skin conditions, and the use of rosehip concentrations in skincare formulations (such as toner, serum, lotion, and sunscreen). Up-to-date studies have revealed that rosehip extracts are perfect candidates as topical application products in the form of nanoemulsions. Extensive in vivo studies have revealed that rosehip extracts also exhibit specific activities against multiple skin disorders (i.e., wound healing, collagen synthesis, atopic dermatitis, melasma, and anti-aging effects). Overall, with multiple dermatological actions and efficacies, rosehip extracts and oils are promising agents that require a thorough investigation of their functioning processes to enable their safe use in the skincare industry.

Supercritical fluid extraction of ginger herbal dust bioactives with an estimation of pharmacological potential using in silico and in vitro analysis

Extensive filter tea production in the last decade has increased the amount of generated waste. Due to the particle size (less than 0.315 mm), this material, also known as herbal dust, cannot be packed into the filter tea bags. According to previous studies, valuable bioactive compounds have been recovered from herbal dusts. Therefore, this study is focused on the valorization of ginger (Zingiber officinale) herbal dust, which is commonly used for the treatment of digestive disorders, colds, and various health issues. To obtain highly valuable extracts rich in non-polar and low-polar bioactive compounds novel Supercritical carbon dioxide (SFE-CO2) extraction was applied. As a reference technique, Soxhlet extraction (methylene chloride) was employed. The chemical profiles of all extracts were determined by GC-MS. The most dominant compounds, according to the results, were hydrocarbon sesquiterpene zingiberene (7.51%–14.28%), sesquiterpenes β-sesquiphellandrene (3.70%–7.34%), β-bisabolene (3.54%–6.93%), ar-curcumene (3.02%–6.19%) and non-volatile phenolic compounds 6-gingerol (25.33%–31.11%), 6- shogaol (12.21%–14.91%). Based on the chemical profile of the extracts, the in silico study was carried out where the effects of ginger compounds with a focus on diabetes were specified. Besides, the most potential SFE-CO2 extract was selected for the investigation of effects on short- and long-term cell viability on liver cancer cells.

Toxicological and Sedative Effects of Chipilin (Crotalaria longirostrata) Leaf Extracts Obtained by Maceration and Supercritical Fluid Extraction

Toxicological and Sedative Effects of Chipilin (<i>Crotalaria longirostrata</i>) Leaf Extracts Obtained by Maceration and Supercritical Fluid Extraction

Optimizing the Supercritical Carbon Dioxide Extraction of Hibiscus Flower Essential Oil using Response Surface Analysis

The development of Supercritical Fluid Extraction (SFE) has opened the door to the harvesting of plants for a wide range of chemical compounds. This study distilled essential oil from hibiscus flowers utilizing the supercritical CO2 method. Different extraction parameters, including pressure (100-300 bar) and temperature (300-350 K), were studied to visualize how they affected oil recovery. Response surface analysis was used to fine-tune the extraction process. The chemical composition of the recovered oil was analyzed by Gas Chromatography-Mass Spectrometry (GC-MS). According to the findings, 13.11% per 80 g of dry flowers is the ideal oil extracted from Hibiscus flowers, using SFE at 200 bar pressure and 325 K extraction temperature. Six compounds were provisionally identified in the extracted oil from hibiscus flowers under optimum SFE conditions.

Advancement in extraction and characterization techniques of natural dyes from dye yielding plants sources: a review

Purpose Everyone is extremely concerned about environmental protection and health safety due to the rise in living standards. Plant-derived natural dyes have garnered much industrial attention in food, pharmaceutical, textile, cosmetics, etc. owing to their health and environmental benefits. The present study aims to focus on the elimination of the use of synthetic dyes and provides brief information about natural dyes, their sources, extraction procedures with characterization and various advantages and disadvantages. Design/methodology/approach In producing natural colors, extraction and purification are essential steps. Various conventional methods used till date have a low yield, as these consume a lot of solvent volume, time, labor and energy or may destroy the coloring behavior of the actual molecules. The establishment of proper characterization and certification protocols for natural dyes would improve the yielding of natural dyes and benefit both producers and users. Findings However, scientists have found modern extraction methods to obtain maximum color yield. They are also modifying the fabric surface to appraise its uptake behavior of color. Various extraction techniques such as solvent, aqueous, enzymatic and fermentation and extraction with microwave or ultrasonic energy, supercritical fluid extraction and alkaline or acid extraction are currently available for these natural dyes and are summarized in the present review article. Originality/value If natural dye availability can be increased by the different extraction measures and the cost of purified dyes can be brought down with a proper certification mechanism, there is a wide scope for the adoption of these dyes by small-scale dyeing units.

Advanced sample preparation and chromatographic techniques for analyzing plant-based bioactive chemicals in nutraceuticals

Since ancient times, humanity has used plants to cure diseases or maintain health. Nowadays, obtaining the bioactive components of plants and researching their effects continues at great speed. Plant-derived chemicals are studied with great interest in the food, pharmaceutical and cosmetic industries. The complexity of city life limits access to plants in their natural habitat. Nutraceuticals are indispensable products of our age to protect health. Many methods have been developed and successfully applied for extracting crude plant extracts and pure active substances included in nutraceuticals from plant tissues and their quantitative analysis. This study compiled the methods for obtaining plant extracts and active ingredients and the techniques developed for analyzing bioactive compounds. The extraction methods of plant-based bioactive chemicals (maceration, Soxhlet extraction, decoction, infusion, hydro distillation, ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), enzyme-assisted extraction (EAE), pressurized liquid extraction (PLE), and supercritical fluid extraction (SFE)) were explained in detail, and some important plant-based bioactive chemicals and source plants were summarized in table form. Additionally, the studies using different analytical techniques such as high-performance liquid chromatography (HPLC), gas chromatography (GC), thin layer chromatography (TLC), mass spectrometry (MS), and ultra-performance liquid chromatography (UPLC) were discussed. Finally, evaluating its effectiveness in the extraction of different bioactive compounds and the prospects for more environmentally friendly methods alternative to traditional solvents were emphasized. This review covers the latest practices and new trends in analyzing plant-based bioactive chemicals in nutraceuticals over the last three years.

Tetraselmis chuii Edible Microalga as a New Source of Neuroprotective Compounds Obtained Using Fast Biosolvent Extraction.

Tetraselmis chuii is an EFSA-approved novel food and dietary supplement with increasing use in nutraceutical production worldwide. This study investigated the neuroprotective potential of bioactive compounds extracted from T. chuii using green biobased solvents (ethyl acetate, AcOEt, and cyclopentyl methyl ether, CPME) under pressurized liquid extraction (PLE) conditions and supercritical fluid extraction (SFE). Response surface optimization was used to study the effect of temperature and solvent composition on the neuroprotective properties of the PLE extracts, including anticholinergic activity, reactive oxygen/nitrogen species (ROS/RNS) scavenging capacity, and anti-inflammatory activity. Optimized extraction conditions of 40 °C and 34.9% AcOEt in CPME resulted in extracts with high anticholinergic and ROS/RNS scavenging capacity, while operation at 180 °C and 54.1% AcOEt in CPME yielded extracts with potent anti-inflammatory properties using only 20 min. Chemical characterization revealed the presence of carotenoids (neoxanthin, violaxanthin, zeaxanthin, α- and β-carotene) known for their anti-cholinesterase, antioxidant, and anti-inflammatory potential. The extracts also exhibited high levels of omega-3 polyunsaturated fatty acids (PUFAs) with a favorable ω-3/ω-6 ratio (>7), contributing to their neuroprotective and anti-inflammatory effects. Furthermore, the extracts were found to be safe to use, as cytotoxicity assays showed no observed toxicity in HK-2 and THP-1 cell lines at or below a concentration of 40 μg mL-1. These results highlight the neuroprotective potential of Tetraselmis chuii extracts, making them valuable in the field of nutraceutical production and emphasize the interest of studying new green solvents as alternatives to conventional toxic solvents.

Comparison of antimicrobial activities of oregano, lavender, sage, anise and clove extracts obtained by supercritical fluid carbon dioxide extraction and essential oils obtained by hydrodistillation

ABSTRACT The aim of the study was to compare the extraction yields, chemical compositions and antimicrobial activities of essential oils obtained by hydrodistillation (HD) and volatile extracts obtained by supercritical fluid extraction extraction (SFE) from oregano (Origanum onites), lavender (Lavandula angustifolia), sage (Salvia officinalis), anise (Pimpinella anisum) and clove (Syzygium aromaticum). Essential oil compounds (EOCs) of the obtained materials from both techniques (HD and SFE) were determined by GC-MS/FID. Agar well diffusion method was used for the determination of antimicrobial activities on five different foodborne microorganisms (Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli Enterococcus faecalis and Klebsiella pneumoniae). According to the results; extraction yields of SFE were found to be higher than HD for all tested aromatic plants. Active ingredient ratios (carvacrol, camphor, anethole, eugenole and linalol) varied depending on the extraction methods. SFE extracts were higher antimicrobial activity than HD essential oils, except clove extract on E. faecalis.

Anti-inflammatory Evaluation of Cannabidiol from Cannabis sativa L. Obtained Using Supercritical Fluid Extraction and Cannabidiol Extract-based Cream

Purpose: This study evaluated the anti-inflammatory properties of a cannabidiol (CBD) extract obtained by supercritical fluid extraction (SFE) from &lt;i&gt;Cannabis sativa&lt;/i&gt; L. and a cream containing the extract. Methods: The CBD extract was quantitatively analyzed using high-performance liquid chromatography. We used a lipopolysaccharide (LPS)-induced RAW 264.7 cell model of inflammation to determine the anti-inflammatory effect of the CBD extract by measuring nitric oxide (NO) production. We evaluated the anti-inflammatory properties of a cream containing the CBD extract by measuring interleukin-1 alpha (IL-1α) levels induced by sodium dodecyl sulfate (SDS) in KeraSkin&lt;sup&gt;TM&lt;/sup&gt;, a human skin model. The cytotoxicity of the CBD and CBD-containing cream was assessed by MTT assay in each model. Results: The average purity of the CBD extracted and refined by SFE was 99.40%. The CBD extract and CBD cream showed concentration-dependent inhibition of inflammation. At 4 μg/mL, NO production in the RAW 264.7 cell model was significantly decreased by 64.98% compared with the negative control. IL-1α production induced by SDS in the KeraSkin&lt;sup&gt;TM&lt;/sup&gt; model decreased by 54.17% in the group treated with cream containing 0.4% CBD compared with the negative control and 22.09% compared with the positive control groups. Conclusion: The medicinal use of CBD is attracting interest in Korea. Our study confirmed its exceptional anti-inflammatory properties and effectiveness in a cream product. Should additional research confirm its safety and efficacy, CBD is expected to replace existing anti-inflammatory ingredients.

Extraction processes of high added value molecules from wood waste: overview and comparison of processes for an eco-friendly recovery from paper industry by-product

The work aims to optimize an industrial greener extraction process of chemical components in wood waste from the paper industry. Extracts from a mixture of two species (Picea abies and Abies alba) were studied. Several parameters were found to play a key role in the extraction yield: to the conventional parameters such as the chemical composition of the extracting medium or the temperature, a key criterion was the choice of a process procedure in order to take into account the absorption of the solvent by the wood. The implementation of several operating modes, namely solid-liquid single-batch extraction, countercurrent extraction, percolation extraction, near supercritical, and supercritical fluid extraction is discussed in terms of yield and ease of implementation when scaling up.

Extension of ultrafiltered cheese shelf life using edible coatings containing supercritical rosemary, thyme and coriander extracts as antimicrobial agents

Currently, supercritical extraction allows for the “green” isolation of plant antimicrobial components and their subsequent use in the food industry. The study aims to extend the shelf life of ultrafiltered cheese by using edible coatings containing supercritical fluid extracts (SFEs) of Rosmarinus officinalis L., Thymus vulgaris and Coriandrum sativum L. The SFEs chemical profiling was studied using GC-MS. Ten emulsion coating formulations were prepared using whey protein concentrate and the SFEs. The antimicrobial activity of coatings was assessed using well diffusion assay. Their effect on shelf life, physicochemical and sensory characteristics of the ultrafiltered cheese samples was investigated. GC/MS analysis of Rosmarinus officinalis L., Thymus vulgaris and Coriandrum sativum L. showed that 1,8-Cineol, m-Cymene, and Linalool are the major compounds with 35.4, 41.6, and 61.3 % respectively. The particle size of all formulations ranged from 157.3 to 453.7 nm and had a broad size distribution with a low Polydispersity Index values ranging from 0.35 to 0.49. The obtained Zeta potential values from −27.8 to −36.0 mV indicate the stability of the SFE-emulsion formulations. The antimicrobial trials showed that formulations, that mainly contained Thymus vulgaris, demonstrated more inhibition effect against all tested strains than other formulas with inhibition zone diameter ranged between 10.7 and 16.3 mm and followed by formulations, that mainly contained Rosmarinus officinalis L., with inhibition zones diameter between 6.7 and 11.7 mm depending on microbial type. Combining rosemary and coriander extracts synergistically enhanced their antimicrobial effect and extended the shelf life of cheese beyond 40 days.

Re-Valorization of Red Habanero Chili Pepper (Capsicum chinense Jacq.) Waste by Recovery of Bioactive Compounds: Effects of Different Extraction Processes

Inadequately managed agricultural waste significantly impacts the environment, health, and economy. This pollution stems from the underutilization, inadequate awareness, and insufficient treatment of agricultural waste. Fruit and vegetable wastes are valuable sources of bioactive compounds. This study aimed to revalorize discarded waste from red habanero chili peppers (Capsicum chinense Jacq.) by extracting bioactive compounds through different extraction processes: maceration (ME), maceration assisted by ultrasound (US), Soxhlet extraction (SE), supercritical fluid extraction (SFE), and supercritical fluid extraction with a co-solvent (SFEC). The extraction processes had significant effects on extraction efficiency and phytochemical profile (capsaicinoids and carotenoids recovery). The results indicated that the highest-efficiency process was SFEC, in addition to its high phytochemicals recovery (14.9 mg of total capsaicinoids and total carotenoids 292.09 µg per gram of sample). Concerning the phytochemical profile of the extract, the maceration process yielded the highest concentration of compounds, followed by US and SFEC. These data reveal that the use of the SFE and SFEC processes is recommended for extracting phytochemicals with biological activity from red habanero chili pepper waste for diverse industrial applications.

Green Extraction Approach for Isolation of Bioactive Compounds in Wild Thyme (Thymus serpyllum L.) Herbal Dust—Chemical Profile, Antioxidant and Antimicrobial Activity and Comparison with Conventional Techniques

The aim of this study was to provide a chemical profile and determine the antioxidant and antimicrobial activity of the essential oil (EO) and lipid extracts of Thymus serpyllum L. herbal dust obtained via conventional (hydrodistillation (HD) and Soxhlet extraction (SOX)) and novel extraction techniques (supercritical fluid extraction (SFE)). In addition, a comparative analysis of the chemical profiles of the obtained EO and extracts was carried out, as well as the determination of antioxidant, antibacterial and antifungal activity of the lipid extracts. According to the aforementioned antioxidant and antimicrobial activities and the monoterpene yield and selectivity, SFE provided significant advantages compared to the traditional techniques. In addition, SFE extracts could be considered to have great potential in terms of their utilization in the pharmaceutical and cosmetic industries, as well as appropriate replacements for synthetic additives in the food industry.

Supercritical fluid and pressurized liquid extraction of spent tucumã-do-Amazonas (Astrocaryum aculeatum) almonds

Supercritical fluid (SFE) and pressurized liquid (PLE) extraction were applied to tucumã-do-Amazonas (Astrocaryum aculeatum) almonds (TAA). SFE was performed with CO2 at different pressures and temperatures, and the extracts were characterized for global yield, fatty acid and triacylglycerol profile, carotenoids, tocols concentration and thermal behavior. PLE was applied to the SFE-defatted almond at different temperatures and ethanol:water mixtures. The PLE extracts were characterized for global yield, total phenolics and antioxidant capacity. SFE global yield was 33 g extract/100 g dry TAA at 25 MPa and 40 °C, containing mainly triacylglycerols from lauric and myristic acids. Regarding tocols, SFE at 25 MPa and 50 °C resulted in 58 µg/g extract. PLE from defatted TAA achieved 24 g extract/100 g defatted TAA and the greatest phenolic content and antioxidant capacities with 75% ethanol at 110 °C. Therefore, TAA is a potential source of valuable compounds to be extracted by SFE and/or PLE.

Preliminary investigation of the safe dose of oleandrin when administered orally to Beagles.

The purpose of the study reported herein was to determine the dose of oleander extract and oleandrin (the key pharmacologically active constituent) that could be safely administered PO to dogs. 42 purebred Beagle dogs were used to study an extract of Nerium oleander. 3 studies were performed in 42 purebred young adult (ages 12 months or older) Beagle dogs using a supercritical fluid extract of N oleander leaves. The first study was an 8-day initial dose-ranging study in 2 dogs, a second 7-day repeat-dosing study was performed in 4 dogs, and the final study was performed in 32 dogs where test subjects were given extract or placebo once daily for 28 consecutive days via oral (gavage) administration followed by a 14-day recovery period. At 2.3 µg/kg of oleandrin, there were no observable adverse effects during the duration of the study. Adverse effects were not seen until doses exceeded 6.9 µg/kg of oleandrin, at which time mild, reversible clinical signs were noted. However, a dose > 460 µg of oleandrin/kg was fatal in 1 of 2 dogs in this study. The studies reported here, taken in totality, suggest that doses exceeding 6.9 µg/kg of oleandrin may be associated with cardiac abnormalities. An estimated no treatment effective adverse event oral dose of oleandrin appears to be 4.6 µg of oleandrin/kg. Higher doses may be tolerable but should be used with appropriate monitoring.

Silicon's Influence on Polyphenol and Flavonoid Profiles in Pea (Pisum sativum L.) under Cadmium Exposure in Hydroponics: A Study of Metabolomics, Extraction Efficacy, and Antimicrobial Properties of Extracts.

The current study aimed to investigate the impact of silicon (Si) supplementation in the form of Na2SiO3 on the metabolome of peas under normal conditions and following exposure to cadmium (Cd) stress. Si is known for its ability to enhance stress tolerance in various plant species, including the mitigation of heavy metal toxicity. Cd, a significant contaminant, poses risks to both human health and the environment. The study focused on analyzing the levels of bioactive compounds in different plant parts, including the shoot, root, and pod, to understand the influence of Si supplementation on their biosynthesis. Metabolomic analysis of pea samples was conducted using a targeted HPLC/MS approach, enabling the identification of 15 metabolites comprising 9 flavonoids and 6 phenolic acids. Among the detected compounds, flavonoids, such as flavon and quercetin, along with phenolic acids, including chlorogenic acid and salicylic acid, were found in significant quantities. The study compared Si supplementation at concentrations of 1 and 2 mM, as well as Cd stress conditions, to evaluate their effects on the metabolomic profile. Additionally, the study explored the extraction efficiency of three different methods: accelerated solvent extraction (ASE), supercritical fluid extraction (SFE), and maceration (MAC). The results revealed that SFE was the most efficient method for extracting polyphenolic compounds from the pea samples. Moreover, the study investigated the stability of polyphenolic compounds under different pH conditions, ranging from 4.0 to 6.0, providing insights into the influence of the pH on the extraction and stability of bioactive compounds.

Comparing Quality and Functional Properties of Protein Isolates from Soybean Cakes: Effect of De-Oiling Technologies

Driven by growing concerns about food supply and the environment, research on alternative protein sources has become increasingly important. In this context, de-oiled seed cakes, particularly soybean cakes, have emerged as a promising option. However, the conventional methods, such as organic solvent extraction, from which these cakes are obtained present several limitations. This study aims to evaluate the efficiency of supercritical fluid extraction (SFE) as an alternative method for de-oiling soybean seeds and obtaining related protein isolates. By using SFE for de-oiling, it was possible to achieve 19% more protein isolates from soybean cakes than the conventional de-oiling method using hexane. Moreover, protein isolates from the SFE de-oiled cake reported significantly improved (p &lt; 0.05) emulsifying abilities and water absorption capacity. Gel electrophoresis and differential scanning calorimetry indicated the presence of a higher concentration of proteins in their native state in the SFE de-oiled flour. Finally, results from the sulfhydryl group content, surface hydrophobicity, and protein dispersibility index also supported these conclusions. The SFE process produced de-oiled soybean cakes with superior functional characteristics and lower environmental impact. Thus, this study provided important information for the food industry to develop more sustainable and healthier production methods.

Effects of different preparation methods on the physical, chemical and functional properties of protein powders from Skipjack tuna (Katsuwonus pelamis) liver

Tuna livers (TL), which are often discarded as waste, are a valuable source of protein for human consumption. However, the preparation method used affects the nutritional and functional characteristics of protein powders. This work aimed to investigate the effects of different preparation methods on the physical, chemical, and functional properties of tuna liver protein powders (TLPPs) following heat (H) treatment, heat and ultrasound-assisted (HU) extraction, alkaline pH shift (APS) process, and supercritical carbon dioxide fluid (SC-CO2 ) extraction. H at 85°C (H85), HU at 80 kHz and 100 W (HU-80-100), APS at pH 11.5 (APS 11.5), and SC-CO2 at 350 bars (SC-CO2 -350) resulted in the remarkably highest total protein content among the different preparation conditions. All TLPPs, except for APS 11.5, showed lighter color characteristics. The most abundant amino acids in all TLPPs were glutamic acid, aspartic acid and alanine. The protein solubility and foaming capacity were efficiently improved by SC-CO2-350. Nevertheless, the emulsion properties and oil holding capacity were greatly enhanced by H85 and HU-80-100, and a significant foaming stability and water holding capacity were found in APS 11.5. Therefore, the TLPPs obtained following different preparation methods are unique and could be potentially utilized as a source of protein ingredients in several food systems.

The molecular mechanism of action for the potent antitumor component extracted using supercritical fluid extraction from Croton crassifolius root

Ethnopharmacological relevanceThe root of Croton crassifolius has been used as a traditional Chinese medicine (TCM), called Radix Croton Crassifolius, and commonly known as “Ji Gu Xiang” in Chinese. Its medicinal value has been recorded in several medical books or handbooks, such as “Sheng Cao Yao Xing Bei Yao”, “Ben Cao Qiu Yuan” and “Zhong Hua Ben Cao”. It has been traditional employed for treating sore throat, stomach-ache, rheumatism and cancer. Aim of the studyAt present, there are limited studies on the evaluation of low-polarity extracts of roots in C. crassifolius. Consequently, the aim of this study was to evaluate the antitumor effect of the low-polarity extract of C. crassifolius root. Materials and methodsExtracts were obtained by supercritical fluid extraction. The extracts were tested for antitumor effects in vitro on several cancer cell lines. A CCK-8 kit was used for further analysis of cell viability. A flow cytometer and propidium iodide staining were used to evaluate the cell cycle and apoptosis. Hoechst staining, JC-1 staining and the fluorescence probe DCFH-DA were used to evaluate apoptotic cells. Molecular mechanisms of action were analyzed by quantitative RT‒PCR and Western blotting. Immunohistochemistry was used for the evaluation of xenograft tumors in male BALB/c mice. Finally, molecular docking was employed to predict the bond between the desired bioactive compound and molecular targets. ResultsEleven diterpenoids were isolated from low-polarity C. crassifolius root extracts. Among the compounds, chettaphanin II showed the strongest activity (IC50 = 8.58 μM) against A549 cells. Evaluation of cell viability and the cell cycle showed that Chettaphanin II reduced A549 cell proliferation and induced G2/M-phase arrest. Chttaphanin II significantly induced apoptosis in A549 cells, which was related to the level of apoptosis-related proteins. The growth of tumor tissue was significantly inhibited by chettaphanin II in experiments performed on naked mice. The antitumor mechanism of chettaphanin II is that it can obstruct the mTOR/PI3K/Akt signaling pathway in A549 cells. Molecular docking established that chettaphanin II could bind to the active sites of Bcl-2 and Bax. ConclusionsTaken together, the natural diterpenoid chettaphanin II was identified as the major antitumor active component, and its potential for developing anticancer therapies was demonstrated for the first time by antiproliferation evaluation in vitro and in vivo.