Min Static Time Research Articles

Accelerated Solvent Extraction of Phenols from Lyophilised Ground Grape Skins and Seeds

The efficient extraction of phenols from grapes is an important step for their reliable quantification. The aim was to optimise the lyophilisation process and the extraction of phenols from grape skins and seeds. The phenol extraction yield from lyophilised tissues was investigated with different accelerated solvent extraction (ASE) operating conditions. Skins and seeds were separated from frozen berries and lyophilised without being ground. The weight loss during lyophilisation was followed daily. Phenols were extracted from lyophilised, cryo-ground seeds and skins with ASE at room temperature and 10.3 MPa using 80% aqueous acetone and 60% aqueous methanol. The effects of ASE operational parameters (the number of extraction cycles (ECs) and static time (ST) duration) were investigated. The yield of extracted phenols was evaluated spectrophotometrically by determining total phenolic index at 280 nm (TPI). The weight of skins and seeds significantly dropped after 24 h of lyophilisation and continued to decrease, although not significantly, up until the 9th day. The optimal lyophilisation time was estimated to be 3 days and 5 days for skins and seeds, respectively. The phenol extraction yield was significantly affected after changes of ASE conditions. Based on TPI, the optimal ASE conditions were as follows: (i) lyophilised seeds—eight ECs with 10 min ST using aqueous acetone and then four ECs with 20 min ST using aqueous methanol; (ii) lyophilised skins—eight ECs with 1 min ST using aqueous acetone and then one EC with 20 min ST using aqueous methanol.

Pressurized Liquid Extraction as a Novel Technique for the Isolation of Laurus nobilis L. Leaf Polyphenols.

Laurus nobilis L., known as laurel or bay leaf, is a Mediterranean plant which has been long known for exhibiting various health-beneficial effects that can largely be attributed to the polyphenolic content of the leaves. Pressurized liquid extraction (PLE) is a green extraction technique that enables the efficient isolation of polyphenols from different plant materials. Hence, the aim of this research was to determine optimal conditions for PLE (solvent, temperature, number of extraction cycles and static extraction time) of laurel leaf polyphenols and to assess the polyphenolic profile of the optimal extract by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) as well as to evaluate the antioxidant activity determined by FRAP, DPPH and ORAC assays. The optimal PLE conditions were 50% ethanol, 150 °C, one extraction cycle and 5 min static time. The polyphenolic extract obtained at optimal PLE conditions comprised 29 identified compounds, among which flavonols (rutin and quercetin-3-glucoside) were the most abundant. The results of antioxidant activity assays demonstrated that PLE is an efficient green technique for obtaining polyphenol-rich laurel leaf extracts with relatively high antioxidant activity.

Targeted analysis of sea buckthorn oil extracted by accelerated solvent extraction technique using green and conventional solvents

Sea buckthorn oil (SBO) was extracted using accelerated solvent extraction with two green solvents (ethanol and 2-methyltetrahydrofuran), and two petroleum-based solvents (hexane and diethyl ether) at four different temperatures (60, 90, 120 and 150 °C), using one extraction cycle at 103.42 bar, 10 min static time, 5 min preheating and 30% flushing volume. β-carotene and α-tocopherol were quantified jointly using HPLC-UV/VIS and the fatty acid profile was analyzed using GC-MS. Extraction yield was also measured and the data was compared to a conventional extraction using Soxhlet. The extraction with ethanol achieved significantly higher concentrations of β-carotene in the final oil (1.67 mg/g oil) compared to the results of any other solvent (e.g. 1.23 mg/g oil for hexane ASE) or to the conventional extraction (1.27 mg/g oil). However, the use of green solvents led to the dragging of unwanted polar compounds (sugars), leading to lower oil yield values after purification.

Accelerated Solvent Extraction of Antioxidant Compounds from Gardeniae Fructus and Its Acetylcholinesterase Inhibitory and PC12 Cell Protective Activities.

Gardeniae fructus is a common neuroprotective medicinal food in China, however the extraction efficiency and mixture activities are rarely mentioned. In this study, accelerated solvent extraction (ASE) parameters were optimized by a response surface methodology to extract antioxidants from Gardeniae fructus. Neuroprotective activity was evaluated using H2O2 and amyloid-β25–35 peptide-treated PC12 cells. By comparing with three other extract methods (i.e., heated refluxing extraction (HRE), ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE)), it was found that the yield (35.10%), total iridoids (27.69%), total flavonoid (6.12%) content, antioxidant activities (IC50 on DPPH, 164.46 µg/mL; FRAP value 4703.54 μmol/L), and acetylcholinesterase inhibitory ability (IC50 92.58 µg/mL) of ASE extract under the optimal condition (150 °C temperature, 10 min static time, 60% ethanol, 2 extract cycles) were significantly higher than other extract methods. The strongest ability to protect PC12 cells from damage was also present in ASE extract, as evidenced by decreasing lactate dehydrogenase and malondialdehyde levels, elevating superoxide dismutase and glutathioneperoxidase activities. Compositional analysis indicated that the extremely high crocetin level in ASE extract (1.30 μg/mg) may offer great potential. Our results indicated that ASE is a proper extraction method that could offer great potential for finding the neuroprotective ability of Gardeniae fructus for the treatment of AD.

Polycyclic Aromatic Hydrocarbon (PAHs) Analyses in Marine Tissues Using Accelerated Solvent Extraction (ASE) in Tandem with In-Cell Purification and GC‑MS

The aim of this research was the replacement of conventional sample extraction techniques for polycyclic aromatic hydrocarbons (PAH) in tissue samples for a reliable, fast and ecofriendly procedure. The method was developed using a pressurized solvent extraction method and assessing two different standard reference materials (fish and mussel) and freeze-dried and fortified sardine samples (Sardinella sp.). Five different extraction procedures were evaluated and the best performance comprised 1 g of lyophilized tissue, 5 g of deactivated (5%) silica, a dichloromethane:methanol (4:1 v/v) mixture, a temperature of 80 °C, three cycles, 10 min of static time and 90 s of purge time. The method selected following these tests was further validated through the analysis of nine replicates of the National Institute of Standard and Technology (NIST) reference material No. 2976, resulting in an effective recovery of 83 ± 14%. The means and uncertainties attained for each PAH were equivalent to those of the reference material, corroborating the reliability of the developed method. A shorter processing time, less use of solvents and reagents and lower extract manipulation produced an effective method aligned with green-chemistry guidelines.

Extraction of Brazil nut kernel oil using green solvents: Effects of the process variables in the oil yield and composition

AbstractThe extraction efficiency of isopropyl alcohol and ethyl alcohol was studied, and the effects of the process variables, static time (St), solvent‐to‐feed (S/F) ratio, and temperature (T) in the oil extraction yield (X0), free fatty acids (FFA) content, total phenolic content (TPC), and β‐sitosterol content were statistically investigated. The amount of solvent used (S/F) did not influence theX0, but St andTsignificantly influenced (p< .05) theX0. The condition that maximized theX0(60.40%) occurred when a St of 9 min and aTof 86°C were used, and low level of FFA were observed in all oils obtained. The process variables studied did not significantly influence (p> .05) in the content of FFA (0.25–0.28%), TPC (29.10–38.47 mg GAE/kg oil), and β‐sitosterol content (9.51–40.32 mg/100 g oil). Its oil is a high source of unsaturated fatty acid. Pressurized liquid extraction is a promising technique for rapid extraction of oil including Brazil nut kernel. Its process recovered 90.58% of oil in 27 min (3 cycles for 9 min of static time).Practical ApplicationsThe optimization of pressurized green solvent extraction has been studied in order to demonstrate that this technology, even employing polar solvents, provides high yields in oil extraction. It is a technology under study that leaves no residue of toxic solvents on vegetable oils. In this study, the use of pressurized liquid extraction with isopropyl alcohol (IPA) and ethyl alcohol (EtOH) as solvents was employed to extract oil from Brazil nut kernel, focusing on the oil rapid recovery as an alternative process to increase the oil yield and improve the use of non‐timber forest product of Amazon biome.

Determination of toxic organic pollutants in fine particulate matter using selective pressurized liquid extraction and gas chromatography–tandem mass spectrometry

Studies investigating toxic organic pollutants in fine dust (PM2.5), such as polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs), are insufficient, despite the pollutants’ potent toxicity. The objective of this study is to develop an analytical method for determining PAHs, OCPs and PCBs in ambient PM2.5 using selective pressurized liquid extraction (SPLE). To maximize the extraction efficiency of target analytes, the extraction parameters of SPLE, particularly solvent type, temperature, static time, and cycle number, were optimized. The highest recoveries were observed under the conditions of dichloromethane:acetone (9:1), 100℃, 5 min of static time, and 1 cycle extraction, which is selected as the optimal method of SPLE. In the method validation, the results showed that the suggested method can quantify 17 PAHs, 20 OCPs, and 63 PCBs in PM2.5. Using urban dust SRM (1648a) and ambient PM2.5 samples, the applicability of the method was also confirmed. Total concentration of PAHs was the highest (2639.42–7377.75 pg/m3) followed by OCPs (80.57–674.69 pg/m3) and PCBs (1.39–9.34 pg/m3). Most of the PAHs were detected, whereas 2–7 compounds among 20 OCPs and 2–6 compounds among 63 PCBs were determined. The developed analytical method is highly efficient in terms of process (a one-step extraction process), time (15 min extraction time per one sample) and solvent usage (less than 30 mL per one sample), showing good performance. This method can be applied to investigate the organic toxicants in PM2.5, and it can contribute to monitoring and risk assessment, leading to an effective risk management policy for PM2.5 in Korea.

Optimization, Bio accessibility of Tricin and Anti-oxidative Activity of Extract from Black Bamboo Leaves

Introduction: Bamboo and black bamboo leaves have long been used as folk remedies for the treatment of hypertension, cardiovascular disease in oriental regions. Many studies have shown that bamboo leaves extract and its main phenolic compounds have antioxidant activity before bio accessibility assessment. Methods: The optimized extraction methods for biological compound tricin by pressuriz ed liquid extraction (PLE) from black bamboo leaves were studied and tricin abundant black bamboo extracts were obtained. Subsequently, the digestive stability, bio accessibility and anti-oxidative activity (total phenolics, DPPH, ABTS) of tricin and tricin abundant extract were examined by in vitro simulated digestion system. Results: The optimized extraction methods for biological compound tricin by pressurized liquid extraction (PLE) from black bamboo leaves were obtained at 200oC, 50% ethanol, 20 min static time, and 425 μm particle size, achieving high extraction efficiency of 249 mg/100 g dry leaves. In simulated digestion process, tricin was unstable in duodenum and jejunum phases but recovery reached 100% after ileum phase and released to the maximum in ileum phase. Total phenolics of the extract was increased at the end of digestion. Moreover, ABTS assay showed the best anti-oxidative activity at the ileum phase with opposite result of DPPH assay. Conclusion: High temperature is beneficial to extract tricin from black bamboo leaves. Tricin was kept stable and the crude extract improved the antioxidant activity during gastrointestinal digestion. The information provided a scientific basis for further study pharmacological activity of tricin and the application of bamboo extract as an antioxidant additive.

Selective fractionation of cholesterol from whole milk powder: optimisation of supercritical process conditions

SummarySupercritical fluid extraction (SCFE) of cholesterol from whole milk powder (WMP) was investigated in this study. The combined effects of temperature (40–80 °C) and pressure (150–250 bar) on the efficacy of cholesterol extraction (mg 100 g−1), modifications in the fat content (FC) (%) and solubility index (SI) (%) of WMP were studied and optimised by the application of response surface methodology (RSM). Variations in the free fatty acids (FFAs) (mg oleic acid per 100 g of milk fat) and lightness value (L*) were also investigated after SCFE process. About 55.8% reduction in cholesterol was achieved at the optimised condition of 68 °C, 207 bar with 40 min static time and 2 h dynamic time at flow rate of 6 L min−1. Extraction at the optimised conditions maximised the yield of cholesterol while retaining the FC, SI, FFA and L* at moderate limits of 23.7%, 85.1%, 7.7 mg per 100 g milk fat and 95.4, respectively.

The solubility of sebacic acid in subcritical water using the response surface methodology

The solubility of sebacic acid in subcritical water was investigated. Response surface methodology was used to study the combined effects of the different parameters such as temperature, static and dynamic time to optimize the process conditions for the maximum solubility of sebacic acid. Solubility results of sebacic acid were observed to be highly correlated with quadratic model according to analysis of variance. The solubility of sebacic acid was found as 500 g L−1 in the optimum conditions at 400 K temperature, 4 min dynamic time and 28 min static time. Approximation models were employed for determining solubility of sebacic acid at elevated temperatures.

Supercritical fluid extraction of uranium and thorium employing dialkyl amides

N,N-dialkyl aliphatic amides with varying alkyl groups viz. N,N-dibutyl-2-ethyl hexanamide (DBEHA), N,N-dibutyl-3,3-dimethyl butanamide (DBDMBA), N,N-dihexyl octanamide (DHOA), N,N-di-sec-butyl pentanamide (DBPA), N,N-dibutyl octanamide (DBOA), have been evaluated for supercritical fluid extraction (SFE) of uranium and thorium from nitric acid medium as well as from tissue paper matrix. The trend in extraction efficiency could be correlated to the structure of the amides and the amide complex of uranium/thorium. Straight chain amide DBOA was found suitable for SFE of both uranium and thorium. Under optimised conditions (200atm. pressure, 323K temperature, 30min static time followed by 30min dynamic time, 0.5M amide in methanol), (94±3)% uranium and (85±4)% thorium could be extracted from 4M nitric acid medium. For tissue paper matrix, under optimised conditions (200atm pressure, 323K temperature, 45min static time followed by 30min dynamic time, 0.2M amide in methanol) (87±7)% uranium and (88±7)% thorium were extractable. For the amide having branching at Cα, DBEHA, extraction efficiency of thorium was as low as 7% for 2M HNO3 medium while extraction efficiency of uranium was still 95%. For tissue paper matrix, extraction efficiency for uranium with 0.2M DBEHA (in methanol) was 86% while only 11% thorium was extractable under same conditions. This fact could be exploited for separation of thorium and uranium.

Determination of lutein from green tea and green tea by-products using accelerated solvent extraction and UPLC.

We aimed to identify the optimum conditions for the extraction of lutein from green tea using accelerated solvent extraction, and achieve improved analytical resolution and sensitivity between lutein and zeaxanthin using an ultra performance liquid chromatography (UPLC) system. The optimized method employed 80% ethanol as the extraction solvent, 160 °C as the temperature, 2 static cycles, and 5 min of static time. In the validation of the UPLC method, recovery was found to be in the range approximately 93.73 to 108.79%, with a correlation coefficient of 0.9974 and a relative standard deviation of <9.29% in inter- and intraday precision analyses. Finally, the lutein contents of green tea and green tea by-products were measured as 32.67 ± 0.70 and 18.18 ± 0.68 mg/100g dw, respectively. Furthermore, we verified that green tea by-products, which are discarded after producing green tea beverages, might be used as a great resource for massive lutein production. We have demonstrated that the common problem of inadequate resolution between lutein and zeaxanthin during carotenoid analyses can be overcome by optimizing the combined techniques of accelerated solvent extraction and ultra performance liquid chromatography (UPLC). UPLC was highly effective for saving time, solvent, and labor, as well as providing better resolution. The results in this study demonstrated that green tea by-products could be used as new sources for industrial lutein production owing to their massive production during the extraction of green tea beverages.

Novel Method to Produce Emulsions Containing Essential Oils from Saponin-Rich Pressurized Aqueous Plant Extracts

Abstract In this work, a novel on-line process for production of emulsions containing essential oils from saponin-rich pressurized aqueous plant extracts in one step was developed. Named by our research group as Emulsion from Pressurized Liquid Extraction (EPLE) the feasibility of this process was demonstrated under optimum PLE conditions (12 MPa, temperature of 393 K and 10 min of static time) using Pfaffia glomerata roots as source of saponin-rich extract, water as extracting solvent and clove essential oil as a model dispersed phase. The optimization study of PLE was performed with extraction pressure (12 MPa), temperature (353-433 K) and static extraction time (5-15 min) as independent variables. Only the extraction temperature significantly influenced the extraction yield and extract emulsification index. As expected, the essential oil/aqueous solution (extract dissolved in water) ratio had a strong influence on the emulsion droplet size (d 32 ), and as this ratio is increased, the d

A one-step extraction/clean-up method for determination of PCBs, PBDEs and HBCDs in environmental solid matrices

A selective pressurized liquid extraction (S-PLE) method was developed for rapid determination of 3 classes of halogenated organic contaminants in indoor dust, soil and sediment samples. The optimised method used 3 : 2 v/v n-hexane-dichloromethane for extraction of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecanes (HBCDs). Extraction was performed at 90 °C for 5 min followed by 4 min static time under 1500 psi. Good recoveries of target analytes were obtained after 3 extraction cycles. In-cell cleanup was performed using 10 g of 44% H2SO4 acid silica and 5 g of florisil (secondary fat retainer), while copper powder was used to remove elemental sulfur. The method was validated using NIST SRM2585 and SRM 1941b in addition to an in-house previously characterised soil sample. Measured concentrations of target compounds showed good agreement with the certified values with RSD < 20% indicating the good accuracy and precision of the S-PLE method. Clean extracts provided low noise levels resulting in low method detection limits (<0.03 ng g(-1)) and LOQs (<0.1 ng g(-1)). The method developed was applied successfully to real environmental samples and it provided various advantages over traditional methods including reduced solvent consumption and analysis time, minimal sample contamination and high sample throughput which can be beneficial for environmental monitoring programs dealing with large numbers of samples.

Use of pressurized hot water extraction and high performance liquid chromatography–inductively coupled plasma–mass spectrometry for water soluble halides speciation in atmospheric particulate matter

The feasibility of pressurized hot water extraction (PHWE) has been novelty investigated to speed up water soluble halide species (bromide, Br−; bromate, BrO3−; iodide, I− and iodate, IO3−) leaching from atmospheric particulate matter (PM10 and PM2.5). Total bromine and iodine and total water soluble bromine and iodine have been assessed by inductively coupled plasma—mass spectrometry (ICP–MS). Water-soluble bromine and iodine species were also measured by ICP–MS after anion exchange high performance liquid chromatography (HPLC). Variables inherent to the pressurized hot water extraction process (temperature, modifier concentration, static time, pressure, number of cycles and dispersing agent mass) were fully studied. Results showed that the pressurized leaching procedure can be performed in 9min (5min for pre-heating, 2min of static time, 1min of purge time, and 1min of end relief time). The use of diluted acetic acid as a modifier did not improve the target recoveries. Dispersing agent (diatomaceous earth) was not needed, which reduces the time for filling the cells. Water-soluble halides were reached under the following extraction conditions: extraction temperature of 100°C, pressure of 1500psi, static time of 2min and 1 extraction cycle. Optimized HPLC conditions consisted of an isocratic elution with 175mM ammonium nitrate plus 15% (v/v) methanol as mobile phase (optimum flow rate of at 1.5mLmin−1). Analytical performances, such as limits of detection and quantification, repeatability and analytical recoveries of the over-all procedure have been established. Results obtained show water soluble halides accounted for approximately 20.9±1.3 and 11.8±0.6% of the total bromine and total iodine, respectively. A 79 and 89% of bromine and iodine was non-water soluble, which may be organic non-water soluble species. Br− and IO3− were found to be the major species, and they accounted for 100% of the total water-soluble bromine and iodine.

Development of a sensitive and robust liquid chromatography coupled with tandem mass spectrometry and a pressurized liquid extraction for the determination of aflatoxins and ochratoxin A in animal derived foods

A liquid chromatography–tandem mass spectrometry (LC–MS/MS) with a pressurized liquid extraction (PLE) was developed for the simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2, and ochratoxin A in the muscle, liver, kidney and fat of swine, bovine and sheep, muscle and liver of chicken, muscle and skin of fish, as well as in hen eggs and dairy milk. Samples were extracted with PLE and cleaned-up with solid phase extraction (SPE) on HLB cartridges. The optimum extraction conditions were obtained as a 11ml ASE cell, acetonitrile/hexane as the extraction solvent, 1500psi, 100°C, a 5min static time and a 60% flush volume. A cheaper and widely used SPE column (Oasis HLB) was applied during clean up. The detection and quantification of the 7 mycotoxins were performed by a reversed-phase liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (LC/ESI-MS/MS). The limits of detection defined as CCα varied from 0.07μg/kg to 0.59μg/kg. The recoveries of spiked samples from 0.25μg/kg to 1μg/kg ranged from 68.3% to 105.7% with the relative standard deviations of less than 17.6%. Performances of the whole analytical procedure met the criteria established by the European Commission for mass spectrometric detection.

Analysis of coenzyme Q10 in bee pollen using online cleanup by accelerated solvent extraction and high performance liquid chromatography

A method for the determination of coenzyme Q10 in bee pollen has been developed applying an online cleanup of accelerated solvent extraction and using environmentally acceptable organic solvents. The extracted samples were analysed by high performance liquid chromatography with diode array detection. The optimised method employed 10mL extraction cells, 1g sample size, absolute ethanol as extraction solvent, 80°C of extraction temperature, one extraction cycle, 5min of static time, Cleanert Alumina-N as sorbent and 60% flush volume. The method was validated by means of an evaluation of the matrix effects, linearity, limit of detection (LOD) and quantification (LOQ), trueness, precision and stability. The assay was linear over the concentration range of 0.25–200mg/L and the LOD and LOQ were 0.16 and 0.35mg/kg, respectively. The recoveries were above 90%. The inter- and intra-day precision was below 6.3%. The method has been successfully applied to the analysis of bee pollen samples. For 20 bee pollen products, the coenzyme Q10 content varied from not detectable to 192.8mg/kg.

Determination of Capsaicin and Dihydrocapsaicin in Some Chilli Varieties using Accelerated Solvent Extraction Associated with Solid-Phase Extraction Methods and RP-HPLC-Fluorescence

Reversed phase-HPLC with fluorescence detection of two major capsaicinoids was described. Isocratic elution using a ratio of methanol and deionized water (66:34, v/v) as mobile phase was used at a flow rate of 0.9 mL/min with well achieved separation within 6 min. Under optimum conditions, their analytical figures of merit for the HPLC method were validated. The linearity was in the range of 1.0-25.0 μg/mL with multiple determination coefficients of higher than 0.995. The limit of detection was ranged of 0.008-0.01 μg/mL. The repeatability and reproducibility of the retention time and peak area for these compounds were in good precision with their relative standard deviations (RSDs) lower than 1% and 5%, respectively. Both capsaicin and dihydrocapsaicin were extracted using an accelerated solvent extraction (ASE) of methanol as an extraction solvent for 5 min static time with 3 cycles. The methanolic extracts were subjected to clean up with C18solid-phase extraction (SPE) with its recoveries ranking of 90.2-98.0%. The method recoveries of real samples were found to be 60.7-98.6%. The optimized extraction method were applied for the determination of the two capsaicinoids in ten vareities of hot chilli pepper samples. Total contents of capsaicinoids were found in the range of 2,307.0-9047.3 μg/g DW with their corresponding Scoville heat unit (SHU) of 34,600-135,700. Additionally, the contents of capsaicinoids using external calibration method comparing with those of standard addition were not significantly different, indicating accuracy of the method. Mostly, the contents of capsaicin found in these real samples were rather higher than those of dihydrocapsaicin.

Pressurized Liquid and Superheated Water Extraction of Active Constituents from Zingiber cassumunar Roxb. Rhizome

Active compounds, terpinene-4-ol and (E)-1-(3,4-dimethoxyphenyl) butadiene (DMPBD), displaying anti-inflammatory activity, were successfully extracted from the rhizome of Zingiber cassumunar Roxb. by pressurized liquid extraction (PLE) using methanol or ethanol and by superheated water extraction (SWE). Optimum conditions of both techniques were 1 mL/min flow rate and 5 min static time. The extraction temperature and dynamic time of the PLE and the SWE were 100°C 5 min and 140°C 20 min, respectively. Compared with water distillation, solvent sonication, and Soxhlet extraction, the PLE produced the highest amount of DMPBD and terpinene-4-ol, whereas the SWE and water distillation provided similar results.

Separation of γ‐linolenic and other polyunsaturated fatty acids from Boraginaceae via supercritical CO2

Essential fatty acids were extracted from Echium amoenum (Boraginaceae) seed oil via supercritical carbon dioxide and the results were compared with conventional Soxhlet method as the base case of 100% recovery. The response surface methodology was used to optimize the effective extraction parameters. The chemical composition of recovered oil was analyzed by polar and non-polar gas chromatograph-flame ionization detector columns. The experimental results indicated that echium seed contained 25 wt% oil and the maximum extraction oil recovery of 92% was obtained via supercritical CO(2) at optimal operating conditions (43°C, 280 bar, 1.5 mL/min, 25 min static time and 130 min dynamic time). At similar operating conditions, applying 2.5 and 5 mol% ethanol as a modifier enhanced the recovery to 96 and 112%, respectively. The results showed that supercritical fluid extraction is a viable technique for separation of constituents such as γ-linolenic acid (7-8%), palmitic acid (6-7%), stearic acid (3-4%), oleic acid (12-13%), linoleic acid (19-20%), α-linolenic acid (40-41%) and stearidonic acid (8-9%) from Boraginaceae.