Investigation of Oleuropein Content in Olive Leaf Extract Obtained by Supercritical Fluid Extraction and Soxhlet Methods

The separation processes of the phenolic compounds from solid plant matrixes are of great importance. In the scope of developing more efficient methods to separate olive leaf extract, dried and ground olive tree leaves from Aegean region of Turkey were extracted by means of Soxhlet and supercritical fluid extraction (SFE) methods. In the Soxhlet method, different types of solvents (hexane, water, ethanol, methanol, and methanol/hexane (3:2, v/v) mixture) were used to determine the effect of the solvent type on the extraction performance. In the SFE method, the effect of pressure (100–300 bar), temperature (50 and 100°C), and type of co-solvent on the amount of both extract and oleuropein were investigated. Ethanol, methanol and water were selected as co-solvent in 20% (v/v) amount. Quantitative analysis was performed by using a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) technique. The results of SFE were compared with those obtained by the Soxhlet method. Whereas the highest oleuropein yield was achieved via the Soxhlet method through methanol with the value of 37.84 mg/g dried leaf, the best oleuropein yield was achieved with the value of 14.26 mg/g dried leaf by using CO₂ modified by methanol at 300 bar and 100°C in the SFE method.     

Enrichment,in vitro,and quantification study of antidiabetic compounds from neglected weed Mimosa pudica using supercritical CO2 and CO2-Soxhlet

Supercritical fluid extraction (SFE) using carbon dioxide (CO₂) and liquid CO₂ using Soxhlet (CO₂-Soxhlet) extraction were employed to extract three (3) antidiabetic compounds viz. stigmasterol, quercetin, and avicularin from Mimosa pudica. Various extraction parameters were studied. Extracts were analyzed pharmacologically, qualitatively and quantitatively to ascertain enrichment levels. All three antidiabetic compounds were effectively enriched under optimized conditions of temperature 60°C, pressure 40 MPa, co-solvent ratio 30%, and CO₂ flow rate of 5 ml min^?1. SFE was found to be the better method for enrichment of the antidiabetic compounds than the CO₂-Soxhlet method. Extraction conditions were seen to affect the enrichment of desired compounds.     

Extraction of bioactive enriched fractions from Eruca sativa leaves by supercritical CO2 technology using different co-solvents

Supercritical fluid extraction from freeze-dried Eruca sativa leaves is assessed with the aim of studying the feasibility to obtain bioactive enriched fractions containing different classes of valuable compounds. Total extraction yields and compositions using pure CO₂ and CO₂ + selected co-solvents are compared. Overall extraction curves, fitted by the model of broken and intact cells developed by Sovová, are reported and the influence of the main parameters that affect the extraction process is analysed. The extract with the highest content in glucosinolates and phenols was collected at 30 MPa and 75 °C using 8% (w/w) of water with respect to the CO₂ flow rate, whereas the fraction richest in lipids was obtained using 8% (w/w) of ethanol as co-solvent at 45 °C and 30 MPa. A process including a first step with supercritical CO₂ extraction using water as co-solvent followed by a second step, where a fraction rich in lipids is extracted using ethanol as co-solvent, is proposed. SCCO₂ results are compared with Soxhlet and other methods that combine organic solvents with ultrasounds.     

Extraction of peanut skin oil by modified supercritical carbon dioxide: Empirical modelling and optimization

ABSTRACT

Peanut skin is a waste by-product from peanut industries. It is rich in antioxidants and bioactive compounds. Therefore, the objective of this study was to empirically model and optimize supercritical CO₂ extraction of oil from peanut skin. The extraction conditions were pressure (100, 200 and 300 bar), temperature (313, 328 and 343 K) and rate of modifier ethanol (0.075, 0.15 and 0.225 mL/min). The extraction process was subsequently examined using modified Brunner and Esquivel models. The optimum conditions for extraction peanut skin oil were 279 bar, 70°C and rate of modifier of 7.5% with a maximum yield of peanut skin oil of 0.83 g and initial slope of 0.568 g/min.     

Phase Behavior of Supercritical CO2 Microemulsion with AOT and its Solubilization Properties of 1,3-propanediol

The phase behaviors of three microemulsion systems were studied in a custom-made volume-variable view cell, including the ternary system of CO₂/EtOH (ethanol)/1,3-PDO (1,3-propanediol), quaternary system of CO₂/EtOH/H₂O/1,3-PDO and quinary system of AOT/CO₂/EtOH/H₂O/1,3-PDO, using 1,3-PDO as the model compound, AOT (sodium bis(2-ethylhexyl) sulfosuccinate) as surfactant and ethanol as co-solvent in the CO₂ continuous phase. The phase behaviors of the AOT/CO₂/EtOH/H₂O/1,3-PDO quinary system were mainly discussed in the temperature range of 29.1°C–44.3°C and pressure range of 6.41 MPa-15.95 MPa. The result shows that a thermodynamically stable microemulsion can be formed by controlling the operating pressures and temperatures, which can provide basic thermodynamics data for industrial design and proper operating conditions for selectively solubilizing 1,3-PDO from dilute aqueous solution.     

A food grade approach for the isolation of major alkylresorcinols (ARs) from rye bran applying tailored supercritical carbon dioxide (scCO2) extraction combined with HPLC

Tailored supercritical carbon dioxide (scCO₂) extraction of alkylresorcinols (ARs) from rye bran resulting in pre-purification of ARs already during extraction process. The pre-purification was achieved by splitting the extraction process in 2 steps._. In both steps, the CO₂ pressure was set at 25 MPa with a flow rate of 10 g/min. The first step was carried out at 70 °C, 0.06% of ethanolic co-solvent for 2 h, followed by the second step at 45 °C using 10% of ethanolic co-solvent for 4 h, resulting in a pre-purified ARs extract. From the pre-purified scCO₂ extract, chromatographed on C8 column, were obtained pure ARs homologues of C17, C19 and C21 (68% of total) eluting separately in a linear gradient of ethanol. Additionally, a rapid method for the isolation of high purified pools of ARs homologues using a disposable solid phase extraction SPE-C18 column and a step gradient of ethanol was developed.     

Study on supercritical extraction of lipids and enrichment of DHA from oil-rich microalgae

The present study aims to isolate the lipids from microalgae by supercritical CO₂ (SC-CO₂) extraction followed by a further enrichment of crude lipids to produce high-purity docosahexenoic acid (DHA) by an urea complexation method. Our systematic approach indicates the optimum conditions of supercritical CO₂ extraction were obtained as follows: 35 MPa, 40 °C, ethanol (95%, v/v) as the co-solvent, and the mass ratio of material to co-solvent 1:1. Under these conditions, 33.9% of lipid yield and 27.5% of DHA content were achieved. Despite the relatively low lipid yield, supercritical CO₂ extraction has exhibited many advantages over the Soxhlet extraction for the DHA enrichment such as high DHA purity and superb product quality. Furthermore, urea complexation method on DHA enrichment considerably increased the DHA purity from 29.7% to 60.4% with an enrichment ratio of 60.6%, under the optimum complexation conditions of urea/fatty acid 2:1, complexation time 8 h, and the complexation temperature of −10 °C.     

Molecular Imprinted Solid-Phase Extraction System for the Selective Separation of Oleuropein from Olive Leaf

Oleuropein has many antimicrobial, antiviral, and anticancer features found in olive leaf. Therefore, its isolation from olive leaf is very important in such kinds of applications. In this study, a solid-phase extraction system based on the molecularly imprinted polymer (MIP) was proposed for the selective separation of oleuropein from olive leaf. First, oleuropein imprinted polymer has been prepared by the suspension polymerization using methacrylolamidoantiprine–iron (III) metal-chelate monomers. After that, the oleuropein adsorption capacity and selectivity of the prepared imprinted polymer has been determined. The maximum adsorption capacity of oleuropein has found to be 140 mg g^?1. Finally, MIP has been used as a sorbent in the solid-phase extraction for the separation of oleuropein from crude extract of olive leaves. The oleuropein analyses have been realized by high performance liquid chromatography. The obtained results indicated that the prepared molecularly imprinted sorbent could be used for at least 10 times for purification of oleuropein from olive leaf. The application of the proposed system in the real sample showed that 24.2 mg pure oleuropein could be obtained from 1.0 g of crude olive leaf extract. As a result, the low cost, simple, and selective adsorbent has been developed for oleuropein adsorption. Supplemental materials are available for this article. Go to the publisher's online edition of Separation Science & Technology to view the supplemental file.     

Guava (Psidium guajava L.) seed oil obtained with a homemade supercritical fluid extraction system using supercritical CO2 and co-solvent

In this work we designed and built a homemade supercritical fluid extraction (HM-SFE) system, in which pure CO₂ and CO₂ with co-solvents were used. The HM-SFE was made by means of thermal dilatation-contraction (TDC). This HM-SFE system was used for obtaining guava (Psidium guajava L.) seed oil, using supercritical CO₂ adding ethanol as co-solvent (CO₂ SC/EtOH), extractions were performed at 313 K and different pressures (10, 20 and 30 MPa), each one in four stages of 30 min, the extract with higher yield was subjected to transesterification and high-resolution gas chromatography (HRGC) analysis. The highest extraction yield was obtained at 30 MPa (17.30% w/w), this yield was higher than one observed in a previous work using SC-CO₂, and near to the one obtained by Soxhlet extraction (20.2% w/w). HRGC enabled the identification of components of the derivatized extract as methyl esters of palmitic, oleic, linoleic, and stearic fatty acids. The results obtained with HM-SFE system was compared with a commercial SFE system, obtained very similar results. In this work was possible to construct a low cost and simple manner HM-SFE system which was employed for obtaining guava seed oil, using CO₂ SC/EtOH.     

Influence of pretreatments for extraction of lipids from yeast by using supercritical carbon dioxide and ethanol as cosolvent

Saccharomyces cerevisiae is one of the most studied and industrially exploited yeast. It is a non-oleaginous yeast whose lipids are mainly phospholipids. In this work, the extraction of yeast lipids by supercritical carbon dioxide (SCCO₂) and ethanol as a co-solvent was studied. In particular our attention was focused on the selectivity toward triglycerides, and in a subsequent extraction of the phospholipids present in the yeast. Indeed CO₂ is a non-polar solvent and is not an efficient solvent for the extraction of phospholipids. However, SCCO₂ can be used to extract neutral lipids, as triglycerides, and the addition of polar co-solvents like ethanol, at different compositions, allows a more efficient extraction of triglycerides, and also an extraction-fractionation of phospholipids. In this work SCCO₂ extractions of a specific membrane complex of S. cerevisiae, obtained from an industrial provider, were carried out at 20 MPa and 40 °C, using ethanol as a co-solvent (9%, w/w). It was shown that different pretreatments are necessary to obtain good extraction yields and have a great impact on the extraction. The kinetic of the extractions were successfully modeled using Sovova's model. From the fitting of the main parameters of the model it was possible to compare the effects of the pretreatments over the yeast material, and to better understand the extraction process. Among the seven tested pretreatments the more appropriate was found to be an acid hydrolysis followed by a methanol maceration.     

Effect of extraction method and extraction solvent on recovery of phenolic compounds from olive leaves in Kemalpaşa-İzmir (Turkey): Oleuropein recovery as a case example

Oleuropein, especially in its leaves, has an important role in durability of olive tree with its protective effect against diseases and pests. In this study, oleuropein has been extracted from olive leaves by using different extraction methods. Effects of extraction solvent and the extraction method that were obtained from Kemalpa?a-?zmir, Turkey, were studied. Results showed that the proposed method and the solvent contained more amount of extracts than existing extraction methods. Efficient extraction without degradation was achieved in 2 h. It has also been found that modified superheated liquid extraction in 50:52:25 ethanol:isopropanol:water composition gave the highest recovery.     

Extracts from pitanga (Eugenia uniflora L.) leaves: Influence of extraction process on antioxidant properties and yield of phenolic compounds

Different extraction processes were employed to extract the polyphenolic compounds from pitanga (Eugenia uniflora L.) leaves: a one-step process using water, ethanol or supercritical CO₂ as solvents, and a two-step process using supercritical CO₂ followed by either water or ethanol. The total polyphenolic compounds, total flavonoids and antioxidant activity were determined in all the extracts obtained. The process performance was evaluated with respect to three variables: global extraction yield, concentration and yield of both polyphenols and flavonoids in the extracts. For the one-step extraction, the results showed that the extraction yield increased with solvent polarity. For the two-step process, the results suggested that water was more efficient in extracting the phenolic compounds from E. uniflora when the matrix was previously extracted with scCO₂. With respect to the antioxidant activity, the ethanolic extracts obtained from both processes, using either the DPPH radical scavenging method or the β-carotene bleaching method, presented high antioxidant activities.     

Supercritical fluid extraction of jojoba oil

Supercritical fluid extraction of jojoba oil from Simmondsia chinensis seeds using CO₂ as the solvent is presented in this study. The effects of process parameters such as pressure and temperature of extraction, particle size of jojoba seeds, flow rate of CO₂, and concentration of entrainer (hexane) on the extraction yield were examined. Increases in the supercritical CO₂ flow rate, temperature, and pressure generally improved the performance. The extraction yield increased as the particle size decreased, indicating the importance of decreasing intraparticle diffusional resistance. The maximum extraction yield obtained was 50.6 wt% with a 0.23-mm particle size and a 2 mL/min CO₂ flow rate at 90°C and 600 bar. Use of an entrainer at a concentration of 5 vol% improved the yield to 52.2 wt% for the same particle size and also enabled the use of relatively lower pressure and temperature, i.e., 300 bar and 70°C.     

Comparison of different extraction methods for the extraction of major bioactive flavonoid compounds from spearmint (Mentha spicata L.) leaves

Different bioactive flavonoid compounds including catechin, epicatechin, rutin, myricetin, luteolin, apigenin and naringenin were obtained from spearmint (Mentha spicata L.) leaves by using conventional soxhlet extraction (CSE) and supercritical carbon dioxide (SC-CO₂) extraction at different extraction schemes and parameters. The effect of different parameters such as temperature (40, 50 and 60 °C), pressure (100, 200 and 300 bar) and dynamic extraction time (30, 60 and 90 min) on the supercritical carbon dioxide (SC-CO₂) extraction of spearmint flavonoids was investigated using full factorial arrangement in a completely randomized design (CRD). The extracts of spearmint leaves obtained by CSE and optimal SC-CO₂ extraction conditions were further analyzed by high performance liquid chromatography (HPLC) to identify and quantify major bioactive flavonoid compounds profile. Comparable results were obtained by optimum SC-CO₂ extraction condition (60 °C, 200 bar, 60 min) and 70% ethanol soxhlet extraction. As revealed by the results, soxhlet extraction had a higher crude extract yield (257.67 mg/g) comparing to the SC-CO₂ extraction (60.57 mg/g). Supercritical carbon dioxide extract (optimum condition) was found to have more main flavonoid compounds (seven bioactive flavonoids) with high concentration comparing to the 70% ethanol soxhlet extraction (five bioactive flavonoids). Therefore, SC-CO₂ extraction is considered as an alternative process compared to the CSE for obtaining the bioactive flavonoid compounds with high concentration from spearmint leaves.     

SOLUBILITY OF HIPPOPHAE RHAMNOIDES L. SEED OIL IN SC CO2 AND SCFE PROCESS ENHANCEMENT BY ULTRASOUND

Dynamic method was applied to measure the solubility of oil from Hippophae rhamnoides L. seed in supercritical CO₂. The experiments were carried out at pressures and temperatures ranging from 10 to 30 MPa and 303 to 323 K, respectively. No marked changes in composition of extracted oil in the course of extraction were observed. The solubility was correlated with CO₂ density and temperature by adjusting the constants of the del Valle-Aguilera equation. In order to enhance the extraction efficiency of supercritical fluid extraction (SCFE) for natural products, the SCFE process coupled with power ultrasound (20 kHz, 150 W) was investigated. The reactor with a power ultrasound transducer was specially designed and set up. For Hippophae rhamnoides L. seed oil, soybean seed oil, and licorice root oil extraction, the extraction rate and the oil yield were increased by 16.9–33.2% and 30% respectively, with the coupling of SCFE and power ultrasound.     

Application of a Supercritical CO2 Extraction Procedure to Recover Volatile Compounds and Polyphenols from Rosa damascena

Abstract

A supercritical CO₂ (Sc-CO₂) extraction procedure to recover volatile compounds and polyphenols from Rosa damascena is investigated. It consists of two steps: the first by Sc-CO₂ at 16 MPa and 313.15 K and on-line fractionation using two separators (S1: 7 MPa/ 298.15 K; S2: 5 MPa/ 288.15 K) for volatile compounds, the second by Sc-CO₂ added with 10% ethanol-water mixture (57% v/v) at 8 MPa and 313.15 K for polyphenols. Sc-CO₂ extract obtained in S2 resulted of high quality compared with essential oil. Polyphenol yield by SC-CO₂ added with co-solvent resulted about 80 % of methanol extraction (3250 mg GAE/100 g dw).     

Supercritical fluid extracts from tamarillo (Solanum betaceum Sendtn) epicarp and its application as protectors against lipid oxidation of cooked beef meat

The possibility of using the tamarillo (Solanum betaceum (Cav.) Sendtn (syn. Cyphomandra betacea)) epicarp as source of compounds with antioxidant activity in cooked beef meat (CBM) was explored. Extracts from tamarillo by supercritical fluid extraction (SFE) and Soxhlet extraction (SE) were obtained. The SFE was performed using pure CO₂ at different temperatures and pressures (40 and 50 °C; 10, 20 and 30 MPa) and CO₂ added with ethanol (CO₂/EtOH) as co-solvent (2, 5 and 8%, w/w). The SFE kinetics and mathematical modeling of the overall extraction curves (OEC) were also investigated. EtOH and hexane were used in the SE. The antioxidant activity (AA) of extracts was evaluated in CBM as well as the protection against lipid oxidation was determined by measuring lipid hydroperoxides (LHP) and thiobarbituric acid reactive species (TBARS). The extract obtained by SFE with CO₂/EtOH (50 °C/30 MPa and 2% of EtOH) showed the highest AA. In SFE, the co-solvent addition improved considerably the AA and the extraction yield. The extracts obtained by SFE with CO₂/EtOH showed a better AA compared with the synthetic antioxidant TBHQ. The highest yield values were achieved by SE with ethanol (7.7 ± 0.4%) and by SFE with 5% EtOH (1.9 ± 0.1%). The results indicate that extracts of tamarillo epicarp are a potential source of antioxidant compounds.     

Supercritical fluids extraction and anti-solvent purification of carotenoids from microalgae and associated bioactivity

This study investigated co-solvent modified supercritical carbon dioxide extraction of lipids and carotenoids from the microalgal species of Nannochloropsis oculata. Supercritical carbon dioxide (SCCO₂) anti-solvent precipitation of carotenoids from the extracts following purification of Zeaxanthin was also examined. Continuous modification by ethanol of supercritical carbon dioxide extractions showed that the addition ratio was important for extraction efficiency of lipids and carotenoids. SCCO₂ extraction at 350 bar, 323 K and 16.7 wt% of ethanol addition yielded 239.7 mg of triglycerides and 7.61 mg of carotenoids per gram extract with a total yield of 15.5%. SCCO₂ anti-solvent experiments showed that the content of Zeaxanthin in the precipitate was greater than that in the fraction of normal phase column chromatography. The purest Zeaxanthin (93.8%) was then successfully isolated from the purified fraction by using a reverse-phase HPLC column chromatography. Rat macrophages treated by ultra-sonicated water extracts of the SCCO₂ defatted algae showed a positive phagocytotic activity.     

Influence of operation variables on quality parameters of olive husk oil extracted with CO<Subscript>2</Subscript>: Three-step sequential extraction

Supercritical CO₂ extraction is a viable alternative process for the extraction of high-quality oil from olive husk (also known as olive pomace), a residue obtained in the production of olive oil. We analyzed the effect of pressure (100–300 bar), temperature (40–60°C), solvent flow (1–1.5 L/min), and particle size (0.30–0.55 mm) on four important quality parameters of the oil extracted with CO₂: tocopherol concentration, extinction coefficients at 232 and 270 nm, and saponification value. Response surface methodology was used to obtain mathematical expressions related to the operating variables and parameters studied. Results from these experiments were also used to design a three-step sequential CO₂ extraction procedure to obtain a higher-quality extract. The optimal operational sequence consisted of a first extraction step at 75 bar for 1 h using 1% (vol/vol) ethanol modifier, followed by a second extraction stage at 350 bar for 2.5 h without ethanol and a third step, also at 350 bar, for 2.5 h but using ethanol. These extraction conditions obtained an intermediate fraction of oil with 64% yield and all normal parameters according to European Commission food legislation. This fraction is suitable without any further refining. On the contrary, the oils obtained by hexane extraction and by conventional supercritical CO₂ extraction at optimal conditions are suitable for human consumption after further refining. This last finding may result in improved economics of the sequential CO₂ extraction process compared to the conventional extraction method with hexane.     

Oxidative stability of virgin olive oils

An investigation was carried out on virgin olive oils of the Gentile (Larino), Gentile (Colletorto), Coratina, and Leccino varieties, harvested at different times, to assess their oxidation stability. The olive oils were analyzed by means of peroxide, K_232′ and K₂₇₀ values at 1, 6, 12, and 18 mon of storage in green bottles, in the dark, at temperatures ranging from a mean of 6°C in winter to 12°C in summer. A subsample was also oven-tested at 75°C and then analyzed on a weekly basis using the same oxidative parameters. The less ripe the olives (harvested in the same area during 1 mon), the more resistant the olive oils were to forced oxidation. The amount of total phenols in the oils was found to be directly related, even if to a low degree, to the oleuropein content in the olives and inversely related, to the same degree, to (3,4-dihydroxyphenyl)ethanol. The latter is a derivative of oleuropein; (3,4-dihydroxyphenyl)ethanol content increases as the olives ripen, but it is very low in fresh virgin olive oils, owing to the hydrophilic nature of the phenolic alcohol, which goes mainly into the waste-water during processing. Among the varieties considered, Coratina oils showed the highest resistance to forced oxidation because of their high total phenol content.