Supercritical Fluid Extraction Method Research Articles

Metabolic Activity of Anthocyanin Extracts Loaded into Non-ionic Niosomes in Diet-Induced Obese Mice.

Anthocyanins (ACNs) are polyphenols that might reduce pathological processes associated with type 2 diabetes mellitus and other chronic diseases, but their bioavailability is still controversial. In this study, the metabolic activity of oral delivery of ACN-loaded niosomes was investigated and evaluated in a diet-induced obesity (DIO) mice model. ACNs extracted from Vaccinium Meridionale by the supercritical fluid extraction method were loaded in niosomes. The niosomal formulation was physically characterized and further administrated in drinking water to obese, insulin resistant mouse. We evaluated the effect of ACN loaded niosomes on hyperglycemia, glucose and insulin intolerance and insulin blood levels in C57BL/6J mice fed with a high-fat diet. The ACN-loaded particles were moderately monodisperse, showed a negative surface charge and 57% encapsulation efficiency. The ACN-loaded niosomes ameliorated the insulin resistance and glucose intolerance in the DIO mice model. Additionally, they reduced animal weight and plasma insulin, glucose, leptin and total cholesterol levels in obese mice. ACN-loaded niosomes administration, as a functional drink, had a beneficial effect on the reversal of metabolic abnormalities associated with obesity.

Extraction of pumpkin peel extract using supercritical CO2 and subcritical water technology: Enhancing oxidative stability of canola oil

In this study, subcritical water extraction (SWE) and the supercritical fluid extraction (SFE) methods were used for the extraction of pumpkin peel extract. Total phenolic content and carotenoid compounds of extracts were measured. The extracts were added to canola oil at a concentration of 400 ppm and were stored at 30 °C for 60 days. The peroxide, carbonyl and acid values of the oil samples were measured, then compared with 100 ppm of tert-butylhydroquinone (TBHQ) synthetic antioxidants. The results showed that the total phenol content of obtained extract by SFE (353.5 mg GA/100 g extract) was higher than by SWE (213.6 mg GA/100 g extract), while the carotenoid content was higher for obtained extract by SWE (15.22 mg/100 g extract) compared to SFE (11.48 mg/100 g extract). The result of oil oxidation showed that the oxidative stability of the oil containing the mixed extract (SFE–SWE) is higher than the separate extract, consequently showing higher performance in preventing oil oxidation compared to TBHQ.

Effect of melanin reduction by extracts from Ulmus davidiana by supercritical fluid extraction

Nowadays, many people prefer products containing functional substances extracted by using harmless extraction methods from environmentally friendly substances. In addition, according to the aging of the population, it consumes a lot in external parts regardless of gender. Accordingly, in this study, we would like to check the whitening function by applying extracts from Ulmus davidiana by supercritical fluid extraction method to melanocytes (B16F10) cells in order to utilize it as a functional material. Here, USCF was extracted from Ulmus davidiana branch (with barks) by supercritical extraction fluid and USCFR was extracted with 60% alcohol at room temperature and filtered from the residues acquired after supercritical extraction. USCFREA was fractionated with ethyl acetate, and the ethyl acetate layer extract was concentrated under vacuum concentration and freeze-drying to obtain final product. First, it was confirmed that the optimal concentration of DMSO for melanocytes was 1% or less. In addition, by using the extracts from the Ulmus davidiana, the melanin content for extracellular and intracellular was checked, and the best effect of melanin treatment was confirmed in sample 2 treated with 50 µg.

Addition of pumpkin peel extract obtained by supercritical fluid and subcritical water as an effective strategy to retard canola oil oxidation

In the present study, the supercritical fluid extraction (SFE) and subcritical water extraction (SWE) methods were used to obtain the pumpkin peel extract. The obtained extracts were compared together. The results showed that total phenol content in the SFE extract (353.5 mg GA/100 g extract) was higher than the SWE extract (213.6 mg GA/100 g extract). However, carotenoid content was higher in the SWE extract (15.22 mg/100 g extract) compared to the SFE extract (11.48 mg/100 g extract). The extracts were added to canola oil at 400 ppm concentration separately and in combined form (SFE+SWE) and TBHQ was added at 100 ppm. The thermal stability of oil samples at the thermal condition of 180 °C was measured using the Rancimat test, conjugated diene and the polar value. The sample containing the combined SFE and SWE extract had the highest oxidative stability, consequently showing higher performance in preventing oil oxidation during the frying process compared to TBHQ. As a result, pumpkin peel extract can be used as a natural antioxidant in edible oils.

Physicochemical characteristics of Actinostemma lobatum Maxim. kernel oil by supercritical fluid extraction and conventional methods

There are limited studies available regarding the characteristics of Actinostemma lobatum Maxim. kernels. In the present study, the kernels were analyzed for their nutritional composition, especially their oil characteristics. Expressed on the dry weight basis, the kernel oil accounted for 47 % of the kernel, whereas the content of protein represented 26 %. The major unsaturated fatty acids of the kernel oils were linoleic acid and oleic acid (43 %–44 % and 37 %–38 %, respectively). The kernel oils were found to be rich in squalene, tocopherols, and phytosterols (2327–2975, 676–759, and 504–676 mg/kg, respectively). More specifically, δ-tocopherol was the most abundant tocopherol and Δ5-avenasterol was the major sterol in the kernel oils. Furthermore, three different oil extraction methods (supercritical CO2 extraction, Soxhlet extraction, and cold pressing) were applied and the physicochemical properties of the extracted oils were compared. No significant differences (P > 0.05) were found in tocopherol and sterol contents for the three extraction methods. Higher oil yield and oil recovery were obtained by supercritical CO2 extraction. Overall, A. lobatum kernel is a promising valuable source of bioactive substances and essential fatty acids for future applications in food, pharmaceutical, and cosmetic industries.

Evaluation of supercritical fluid extraction for the determination of neonicotinoid pesticides in green onion

A supercritical fluid extraction (SFE) method was presented for the determination of neonicotinoid pesticides in green onion. We optimized the SFE parameters, namely the pressure, temperature, type and concentration of the modifier solvent, and extraction time, by testing on the incurred green onion applied with four commercial neonicotinoid formulations at the harvesting stage. The analytical values of the four neonicotinoids, obtained by 5 min dynamic SFE with a 25% (v/v) methanol/supercritical carbon dioxide mixture at 80 °C and 25 MPa, were in good agreement with those obtained by solid-liquid extraction using a homogenizer. This latter homogenization method is employed as a Japanese official method for the analysis of pesticide residues in food. The SFE-to-homogenization analytical value ratios were in the range 98.8–100.0%. Spike-and-recovery testing with spiked green onion was also performed by optimized SFE. The ratios of the analytical-to-spiked concentrations for six neonicotinoids obtained by the SFE method were 96.1–102.1% for 0.4 mg/kg and 93.9–104.1% for 4.0 mg/kg spiking levels. These values satisfied the Japanese validation guidelines for the testing method of pesticides in food. These results indicate that SFE is applicable for the analysis of neonicotinoids in green onion and enables automatic extraction with a small amount of solvent.

High Density Supercritical Carbon Dioxide for the Extraction of Pesticide Residues in Onion with Multivariate Response Surface Methodology.

The excessive use of pesticides is a serious health problem due to their toxicity and bioaccumulation through the food chain. Due to the complexity of foods, the analysis of pesticides is challenging often giving large matrix effects and co-extracted compounds. To overcome this problem, a selective and “green” supercritical fluid extraction method was developed, using neat carbon dioxide as a solvent at pressures of up to 800 bars. A Box–Behnken response surface experimental design was used, with the independent variables of density (0.70−1.0 g mL−1), temperature (40−70 °C), and volume (10−40 mL) of solvent, and the dependent variable of extracted amount of pesticides. The optimum extraction condition was found at the use of 29 mL of supercritical CO2 at 0.90 g mL−1 and 53 °C (corresponding to 372 bars of pressure). It was observed that increasing the density of CO2 significantly increased the extraction recovery of endrin and 2,4′-dichlorodiphenyldichloroethane. Matrix-matched calibration curves showed satisfactory linearity (R2 ≥ 0.994), and LODs ranged from 0.2 to 2.0 ng g−1. Precision was lower than 11% and recoveries between 80%–103%. Thus, the developed method could efficiently be used for trace analysis of pesticides in complex food matrices without the use of organic solvents.

Effect of Static Extraction Time on Supercritical Fluid Extraction of Bioactive Compounds from Phyllanthus niruri

Supercritical fluid extraction (SFE) is an effective method to extract active components from plants. SFE process can be conducted by using a static or/and dynamic extraction. Static extraction is important for the diffusion of solvated analyte to the matrix surface. Therefore, the main objective of this study is to evaluate the static extraction time influence on SFE extraction of Phyllanthus niruri and the main bioactive compounds. The extraction was conducted by using supercritical carbon dioxide (SC-CO2) with 50% ethanol-water as a co-solvent at selected operating conditions (200 bar, 60 °C, 10% co-solvent content and 3 ml/min solvent total flow rate). The static extraction time was investigated in the range of 15 min–75 min by allowing the plant matrix to immerse in a mixture of SC-CO2 and 50% ethanol-water at the operating condition. The quantification of targeted compounds, which were gallic acid (GA), corilagin (CO) and ellagic acid (EA) were analysed by using high performance liquid chromatography (HPLC). The observation on the physical characteristics of the plant matrix was performed by using the scanning electron microscopy (SEM). The best result for total extraction yield was obtained at 60 min of static extraction time, which was 20.7% g/g sample. It also gave a better result for all three targeted compounds in terms of total content percentage (0.35% g GA/g extract, 3.05% g CO/g extract, and 5.17% g EA/g extract). In conclusion, static extraction time was crucial in extracting bioactive compounds in P. niruri by using SFE method and should be considered as the main parameter other than pressure and temperature.

Supercritical fluid extraction followed by supramolecular solvent microextraction as a fast and efficient preconcentration method for determination of polycyclic aromatic hydrocarbons in apple peels.

In this work, reverse micelle-based supramolecular solvent microextraction method coupled with supercritical fluid extraction and used for determining trace amounts of polycyclic aromatic hydrocarbons in apple peels. The extract was analyzed by high-performance liquid chromatography equipped with a fluorescence detector. Coupling supramolecular solvent microextraction with supercritical fluid extraction method, resolve low preconcentration factor of supercritical fluid extraction method, improved limit of detection of polycyclic aromatic hydrocarbons and allow the use of supramolecular solvent microextraction in solid matrices. The effective parameters on the supramolecular solvent microextraction and supercritical fluid extraction efficiency were optimized using one variable at a time and face centered design methods, respectively. Under the optimum condition, the limits of detection and limits of quantifications were in the range of 0.34-1.27 and 1.03-3.82µg/kg, respectively. Analysis of polycyclic aromatic hydrocarbons in apple peels showed that the supercritical fluid extraction/ supramolecular solvent microextraction method provide great potential for trace analysis of polycyclic aromatic hydrocarbons in fruit samples (RSDs<7.7%).

Effect of temperature and pressure on antimycobacterial activity of Curcuma caesia extract by supercritical fluid extraction method.

Supercritical fluid extraction (SFE) is an advanced technique using CO2as a solvent and plant-based drug exploration is a topic of growing interest. Curcuma caesia is a medicinal herb with many medicinal potential. Hence, in the present study, the effect of temperature (40°C-60°C) and pressure (10-20 MPa) on extraction yield and antimycobacterium potential of C. caesia Roxb. dry rhizome powder using supercritical fluid extraction method were evaluated. The extract of C. caesia by SFE was accomplished using temperature range (40°C-60°C) and pressure range (10-20 MPa). The chemical profile of the extracts were investigated by Gas Chromatography Mass Spectrometry (GCMS) and the antimycobacterial activity of the extracts were analyzed against Mycobacterium smegmatis strains MTCC06 and MTCC994. Compounds found in the extract were further checked by in silico analyses with two protein target 4DRE and 3UCI. Extraction yield ranged from 3.0 to 5.6 g/25g dry substrate, with the highest value being achieved at 50°C and 15 MPa. The results of GCMS analyses revealed the presence of beta-elemene, curzerenone, boldenone, and 2-cyclohexen-1-one, 4-ethynyl-4-hydroxy-3, 5, 5-trimethyl in the extracts. The extract obtained at 50°C temperature and 15 MPa pressure showed the highest zone of inhibition against M. smegmatis strains MTCC06 and MTCC994, that is, 15.6 mm and 13.6 mm, respectively. Active constituents present in the extracts showed good binding energy with 4DRE and 3UCI by in silico analysis. This study identified the effect of temperature and pressure on yield C. caesia extract by SFE method. Furthermore, the effect of different extracts on antimycobacterial potential and docking study validated the antimycobacterial potential.

Comparison of Different Extraction Techniques of Zingiber officinale Essential Oil

Abstract This study aims to find the best conditions for the extraction of Zingiber officinale essential oil using the supercritical fluid extraction (SFE), steam distillation (SD) and hydrodistillation (HD) techniques, regarding the maximum oil yield. For the HD technique is evaluated the best ratio between plant mass and water volume and for SFE and SD the pressure condition was investigated. Principal Component Analysis (PCA) was used to evaluate the similarity between the composition of the essential oil in different pressures and extraction methods. The experimental extraction curve was plotted and three different mathematical models were used to fit the data for SD and SFE methods, obtaining the relevant mass transfer parameters. The essential oil compounds were identified by gas chromatography coupled with mass spectrometry (GC-MS), being α-zingiberene the main component with different contents (from 11.9 to 28.9%). The best condition for the SFE was 100 bar, 40 °C (0.0508 goil/gplant) with 19.34% of α-zingiberene; for the SD, 3 bar (133 °C) (0.00616 goil/gplant) with 28.9% of α-zingiberene; and HD, the volume of 750 mL (0.006988 goil/gplant) with 15.70% of α-zingiberene, all measured on a dry basis.

The Phenolic Compounds, Metabolites, and Antioxidant Activity of Propolis Extracted by Ultrasound-Assisted Method.

The influences of ultrasound-assisted, pharmacopeia, and supercritical fluid extraction methods on bioactive compounds and biological activities of propolis were evaluated. Results showed that propolis extracted by ultrasound-assisted method contained more phenolic compounds, and showed the highest total phenolic content (245.84±6.41mg GAE/g DW), total flavonoids content (198.82±5.74mg RE/g DW), and stronger in vitro antioxidant activity (DPPH·: 1.03±0.04mmol Trolox/g DW, ABTS+·: 2.19±0.05mmol Trolox/g DW, and FRAP: 1.48±0.12mmol FeSO4 /g DW) than those of pharmacopoeia and supercritical fluid methods. A total of 36 phenolic compounds were identified in propolis. Among them, quercetin, quercetin-3-methyl-ether, kaempferol, isorhamnetin, luteolin-methyl-ether, and quercetin-7-methyl-ether could only be found in ultrasound-assisted and pharmacopoeia methods. Moreover, the phenolic compounds had the similar metabolic pathways in rats and were mainly metabolized by sulfation and glucuronidation pathways. Additionally, ultrasonic-treated propolis have good in vivo antioxidant activity and could repair D-galactose-induced oxidative damage in rats. Therefore, ultrasound-assisted method could replace pharmacopeia method to be considered as bioactive compounds extraction from propolis, taking into consideration of yield, short extraction time, and high antioxidant activity.

One-step coextraction method for flavouring soybean oil with the dried stipe of Lentinus edodes (Berk.) sing by supercritical CO2 fluid extraction

Dried Lentinus edodes (Berk.) sing stipe (LES) is mostly discarded owing to its high fibre content and bad palatability, causing significant waste of resources. The aim of this study is to sustainably use LES for the production of flavour soybean oil using the supercritical CO2 fluid extraction (SFE) method. The quality properties of the obtained soybean oil flavoured by LES (OFLS) were evaluated in this study. Profiles obtained with an electronic nose and gas chromatography results obtained by quadrupole time-of-flight mass spectrometry and with a mass-selective detector indicated that mushroom-like aroma compounds were mainly concentrated into the OFLSs from separation kettle A (9 MPa). Interestingly, off-flavour compounds from soybean oil, such as o-xylene and trans-2-nonenal, were mostly distilled into separation kettle B (4 MPa). More importantly, the developed SFE method effectively improved the colour quality of the OFLSs. Additionally, the decrease of the viscosity of the OFLSs indicated that the thermal stability of the edible oil could be improved. In summary, the OFLSs prepared by the green and sustainable SFE method significantly improved the quality of the edible oil.

Evaluation of supercritical carbon dioxide extraction to obtain bioactive compounds from Vernonia amygdalina Delile leaves

Literature reports have shown that supercritical extraction is a promising method to obtain bioactive compounds used to develop drugs based on natural products. This work investigated the yield, antioxidant activity, and phytochemical constituents of the extracts obtained from Vernonia amygdalina Delile leaves with supercritical CO2. The supercritical extraction was examined at temperatures of 40, 50 and 60?C and at pressures of 200 and 250 bar. The supercritical fluid extraction (SFE) method was compared with the classical Soxhlet using different solvents at atmospheric pressure. The overall yield obtained using SFE varied from 0.69 to 1.24%. The extraction conditions which favored the highest yield were 60?C and 250 bar. The overall SFE curves were fitted using three mathematical models. All of them were able to describe satisfactorily the extraction kinetics. Spline model presented the best fit to experimental data and was able to characterize constant and decreasing extraction rate periods. Extracts were characterized by gas chromatography coupled with mass spectrometry technique and some major components were identified. The antioxidant activity tests showed that the SFE extracts had low antioxidant activity exhibiting the estimated concentration of extract required to reduce 50% of the stable free radical DPPH (EC50) values to be between 622.62 and 937.88 ?g mL-1.

Sequential Green Extraction of Caffeine and Catechins from Green Tea

Separation of caffeine and catechins from tea extracts usually requires conventional liquid-liquid extraction employing chloroform. This work was planned to improve a green extraction technique to distunguish these important chemicals from green tea. Extraction of caffeine and catechins from Turkish green tea firstly employing MAE, and then to separate these compounds from each other using a SFE method were purpose of this study. Microwave assisted extraction was applied to extract tea components from green tea then i) conventional liquid-liquid extraction or ii) supercritical carbon dioxide fluid extraction (SFE) method was charged with the effective separation of caffeine and catechins. Initially, an ethanol: water mixture was used in a close microwave system under the particuler extraction situations of green tea samples (fresh, frozen or dried) picked up in three collection periods (first, second and third collection periods). MAE of tea samples was exerted under a controlled 600 W microwave power for 4 min irradiation time at 80 oC temperature. Then MAE crude aqueous extract was divided in to two portions. The first portion was fractionated first with chloroform to distunguish caffeine then ethyl acetate for catechins. Caffeine and catechins were successfully separated. Second portion was freeze-dried and obtained lyophilized solid was used for SFE. Caffeine (3.68% extract yield) was successfully separated from catechins with SFE at 250 bar and 60°C for 180 min. Constituents of the extracts were determined (caffeine and four catechins namely EGC, EC, C, EGCG) by HPLC to evaluate the effectiveness of the separation.

Quickly Identifying High-Risk Variables of Ultrasonic Extraction Oil from Multi-Dimensional Risk Variable Patterns and a Comparative Evaluation of Different Extraction Methods on the Quality of Forsythia suspensa Seed Oil.

Generally, essential oils and components of interest are extracted from plants using organic solvent, distillation, ultrasound and supercritical extraction methods. Ultrasonic extraction (UE) has the advantage of high efficiency, but its process is complicated and it has numerous variables. In this study, an L18-Hunter experimental design was applied for the first time to investigate the practicability of applying UE to Forsythia suspensa seed oil. Six potential high-risk variables, including numerical and non-numeric types, were obtained from the risk analysis and their impacts on global yield and antioxidant activity were screened. Furthermore, oils obtained by different extraction processes (i.e., UE, supercritical fluid extraction (SFE), soxhlet extraction (SE) and hydrodistillation extraction (HD)) were analyzed. A comparative study of these oils was characterized and compared by FT-IR, GC-MS and antioxidant activity. The obtained results show that the type of solvent, solvent-to-solid ratio, extraction power and time were the significant variables affecting the extraction yield, whereas antioxidant activity was only affected by the type of solvent. The regression coefficients of the yield and antioxidant activity models were 0.79 and 0.91, and the ANOVA of the models were 0.013 and <0.0001, respectively. Beta-Pinene was the main abundant component in the oils for the UE, SFE, SE and HD methods and the content was about 46%~52.4%. In conclusion, the L18-Hunter design could be used as an effective experimental design method for rapid screening of high-risk variables. Regarding extraction efficiency, chemical composition and biological activity, UE not only offered a robust Forsythia suspensa seed oil extraction process, but also provided a time- and cost-effective advantage to the food and pharmaceutical industry when compared to the SFE, SE and HD extraction processes.

Supercritical CO2 oil extraction from Bauhinia forficata link subsp. pruinosa leaves: Composition, antioxidant activity and mathematical modeling

The subspecies B. forficata subsp. pruinosa is practically unexplored scientifically. Thus, the aim of this study was to evaluate a supercritical fluid extraction method and to compare it with the conventional Soxhlet method. The supercritical fluid extraction was performed with CO2, at temperatures of 40–60 °C and pressures of 18–22 MPa, while the Soxhlet method was conducted using ethanol, ethyl acetate and n-hexane as solvents. The extract obtained applying the conventional method with ethanol showed the best results for extraction yield, phenolic and flavonoid contents and antioxidant activity. In the case of the supercritical extraction, the highest yield was obtained at 22 MPa and 60 °C. A mathematical model developed by Sovová was applied to describe the experimental extraction curves and they showed a good fit with the experimental data. The major compounds found in the B. forficata extracts were tocopherols, octacosanol and phytol.

Ultrahigh-pressure supercritical fluid extraction and chromatography of Moringa oleifera and Moringa peregrina seed lipids

An ultrahigh-pressure supercritical fluid extraction method was optimized and applied to extract seed oil lipids from two moringa species, namely Moringa oleifera (MO) and Moringa peregrina (MP). A full-factorial design was used to investigate the direct and interaction influence of pressure and temperature in the range of 40 to 80 MPa and 40 to 70 °C, respectively, on the extracted amount of oil from crushed seeds. The results revealed that pressure has a significant positive influence on the extracted amount of oil. The best extraction condition using neat CO2 was found at 80 MPa and 57 °C, yielding 396 ± 23 and 529 ± 26 mg oil per gram of seeds for MO and MP, respectively. An extraction kinetics study revealed a mainly solubility-controlled extraction of oil, and 28 g of CO2 was required to extract 400 mg of oil per gram of seeds of MO using the developed method. Addition of ethanol to the sample prior to the extraction increased the proportion of extractable polar lipids as well as the total amount of extracted oil. The developed method increased the extracted amount of oil twofold compared to a reference method based on solvent sonication. The obtained oil consisted mainly of glycerolipids, sterol esters, and phospholipids. Phospholipids, campesterol, and stigmasterol ester concentrations were found to be higher in MO while cholesterol ester was more abundant in MP.

Installation for the implementation of the supercritical fluid extraction regeneration of nickel-molybdenum catalyst

The process of supercritical fluid extraction recovery of the catalyst at pressures of 10, 20 MPa and temperatures of 343, 373, 433, 473 K with pure supercritical carbon dioxide on the original device has been implemented. An increase in pressure and temperature has a positive effect on the catalyst regeneration process. With increasing pressure increases the density of CO2, and, consequently, its dissolving ability. With increasing temperature, the pressure of saturated vapors of compaction products increases and, as a result, their concentration in CO2 increases. In order to increase the efficiency of the supercritical CO2-extraction process for the regeneration of catalyst samples, carbon dioxide modification with various additives (chloroform, dimethylsulfoxide, acetone, ethanol, and non-polar inert with respect to the catalyst composition hexane) was used. Modification of supercritical carbon dioxide with ethanol and hexane at a temperature of 473 K and a pressure of 20 MPa led to an increase in the mass loss of catalyst samples by 12 and 14%, respectively, which indicate the removal of deactivating substances during regeneration by the method of supercritical fluid extraction.

Xenopus Oocyte's Conductance for Bioactive Compounds Screening and Characterization.

Background: Astaxanthin (ATX) is a lipophilic compound found in many marine organisms. Studies have shown that ATX has many strong biological properties, including antioxidant, antiviral, anticancer, cardiovascular, anti-inflammatory, neuro-protective and anti-diabetic activities. However, no research has elucidated the effect of ATX on ionic channels. ATX can be extracted from shrimp by-products. Our work aims to characterize ATX cell targets to lend value to marine by-products. Methods: We used the Xenopus oocytes cell model to characterize the pharmacological target of ATX among endogenous Xenopus oocytes’ ionic channels and to analyze the effects of all carotenoid-extract samples prepared from shrimp by-products using a supercritical fluid extraction (SFE) method. Results: ATX inhibits amiloride-sensitive sodium conductance, xINa, in a dose-dependent manner with an IC50 of 0.14 µg, a maximum inhibition of 75% and a Hill coefficient of 0.68. It does not affect the potential of half activation, but significantly changes the kinetics, according to the slope factor values. The marine extract prepared from shrimp waste at 10 µg inhibits xINa in the same way as ATX 0.1 µg does. When ATX was added to the entire extract at 10 µg, inhibition reached that induced with ATX 1 µg. Conclusions: ATX and the shrimp Extract inhibit amiloride-sensitive sodium channels in Xenopus oocytes and the TEVC method makes it possible to measure the ATX inhibitory effect in bioactive SFE-Extract samples.