Supercritical fluid extraction of daidzein and genistein isoflavones from soybean hypocotyl after hydrolysis with endogenous β-glucosidases

An optimal condition of supercritical fluid extraction (SFE) for isoflavone aglycones (daidzein and genistein) in soybean hypocotyls previously subjected to thermohydration at pH 5.0 and a temperature of 50 °C for 6, 12 and 18 h was developed. Different temperatures, pressures and cosolvents (methanol, ethanol, and acetonitrile) was tested and compared with solid–liquid extraction using aqueous methanol solution (80% v v⁻¹) conducted in parallel for comparison. The extraction conditions were 50–70 °C, 176–380 bar, adding 0, 5, 10 mol% of cosolvents 80% in water as a modifier. The results from SC–CO₂ showed that the cosolvent and pressure have significant effects in the extraction efficiency. It was found that the extraction conditions promoting the highest extraction of daidzein and genistein were at the temperature of 60 °C, pressure of 380 bar and both static and dynamic extraction of 15 min with the addition of 10% acetonitrile (80% v v⁻¹). The maximum amounts of daidzein and genistein extracted by each method were solid–liquid extraction (70.07 mg 100 g⁻¹) and carbon dioxide–acetonitrile (17.97 mg 100 g⁻¹). The yield of daidzein and genistein achieved by a 30 min SC–CO₂ extraction on soybean hypocotyls after 12 h soaking time was markedly improved by the addition of a modifier (acetonitrile) to the CO₂ fluid. HPLC analysis of the obtained extracts revealed that extraction of isoflavone aglycones by SC–CO₂ was 4.78 and 13.19 mg 100 g⁻¹ for daidzein and genistein, respectively. The contents of daidzein and genistein obtained in the solid–liquid extraction were superior to 86% and 63%, respectively, compared to supercritical extraction.     

Extraction of phenolics from Vitis vinifera wastes using non-conventional techniques

Polyphenols, the well known naturally occurring antioxidants, are the most abundant secondary metabolites in grape wastes. Herein we investigate several non-conventional extraction methods vs classic solid–liquid extraction (SLE) to obtain phenolic compounds from grape seeds and skins. We compared SLE, ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE) and high pressure and temperature extraction (HPTE) in term of extraction yield and antioxidant power of the extract. Solvent of choice between methanol and ethanol was the former, both for skins and seeds. Quali-quantitative analyses were performed using colorimetric and HPLC methods. The highest content in total polyphenols, o-diphenols and flavonoids, both for seeds (108.3, 47.0 mg_GAE g_DW⁻¹, 47.2 mg_CE g_DW⁻¹) and skins (34.2, 10.1 mg_GAE g_DW⁻¹, 21.6 mg_CE g_DW⁻¹) was obtained with HPTE working in a Parr reactor. While the highest antiradical power was determined in seeds extracts from MAE (78.6 μl_extract μg_DPPH⁻¹). Prolonged extraction times (over 30 min) further increased the amount of total polyphenols, while progressively decreased the amount of flavonoids and the antiradical power.     

Effects of parameters on yield for sub-critical R134a extraction of palm oil

Extraction of palm oil from its palm mesocarp using sub-critical 1,1,1,2-tetrafluoroethane (R134a) as an alternative solvent to supercritical CO₂ was studied. The effects of pressure, temperature, flowrate and sample pre-treatment method on extraction yield were examined. Dynamic method was used to determine palm oil solubility at flowrate between 0.5 and 5.0 ml/min. Extraction was performed at pressures between 45 and 100 bar which was lower than those required by CO₂ solvent, and at temperatures between 40° and 80 °C. The results show that extraction yields using R134a increased with pressure and temperature, and that temperature effect (p-value = 0.0000) on palm oil solubility prevailed over that of pressure (p-value = 0.0087). The maximum yield of 66.06 w/w% was obtained at 100 bar and 80 °C which was the best pressure and temperature. Substantial oil yields at relatively lower pressure proved that sub-critical R134a is a viable alternative to CO₂ for extraction of palm oil.     

Supercritical fluid extraction of oil from potato chips: Two scale comparison and mathematical modeling

Oil was extracted from fried chipped potatoes using supercritical carbon dioxide. The goals of the study were to determine the effect of process parameters on the extraction, explore the scalability of the process, and determine useful kinetic parameters. Extraction conditions range 27.6–41.4 MPa (4000–6000 PSI), 35–80 °C and solvent flow rate of 0.5–5.0 g CO₂/min. Up to 100% of the oil was recovered from the potato chips at the highest pressure and temperature conditions. Two process conditions were chosen for comparison of performance with a larger scale (1:5) system, maintaining the same CO₂ flow rate to feed mass ratio. Good agreement between scales was seen at the higher pressure and temperature settings. Kinetic parameters, calculated using a literature model, indicated that, as expected, the extraction is limited by internal diffusion.     

Identification and quantification of polyphenolic compounds from okra seeds and skins

The aims of the present work were to identify and quantify the polyphenolic profile of okra (Abelmoschus esculentus), a vegetable almost worldwide consumed. Since the knowledge about the okra polyphenolic compounds is limited, the seeds and the skins of okra were separately analyzed. The seeds, which represent the 17% of the vegetable and are richer in phenolic compounds, were mainly composed by oligomeric catechins (2.5 mg/g of seeds) and flavonol derivatives (3.4 mg/g of seeds). The skins polyphenolic profile was composed principally by hydroxycinnamic and quercetin derivatives (0.2 and 0.3 mg/g of skins). These findings in associations with the high content of okra in carbohydrates and proteins enhance the importance of this foodstuff in the human diet.     

Inactivation of Saccharomyces cerevisiae in conference pear with high pressure carbon dioxide and effects on pear quality

This work explores the use of high pressure carbon dioxide (HPCD) for the inactivation of Saccharomyces cerevisiae in fresh-cut conference pears. This fruit was chosen as an example of a ready to eat and minimally processed food. Assays were carried out with continuous CO₂ flow at different pressures (6–30 MPa), temperatures (25–55 °C), and exposure times (10–90 min). Heat treatments at similar temperatures and times were compared to the use of HPCD, wherein it was observed that HPCD was more effective. The total inactivation (5 log₁₀ cycles) of the yeast took place at 55 °C with HPCD while it was necessary to reach 70 °C when only heat was applied. Required pressures and exposure times were relatively low (?6 MPa and on the order of minutes) because of the direct contact between the CO₂ and the pear. The pH and °Brix were not affected by the HPCD treatment; however, the pears lost their texture and became darker due to a decrease in vitamin C and enzymatic browning. Peroxidase activity was only partially reduced. The addition of an antioxidant did not help to prevent darkening. Therefore, HPCD could be a low temperature conservation method that is superior to conventional thermal treatments for the preparation of fruit preserves where a firm texture is not essential.     

The optimisation of extraction of antioxidants from potato peel by pressurised liquids

Response Surface Methodology was used to optimise the solid–liquid extraction and Pressurised Liquid Extraction of polyphenols from industrially generated potato peel. Efficiency of extraction was optimised by measuring antioxidant activity, phenol content and the level of caffeic acid. Conditions for optimal antioxidant activity as measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay were 75% ethanol, 80 °C and 22 min with solid–liquid extraction, resulting in an optimum activity of 352 mg Trolox Equivalents/100 g DW potato peel. In comparison, the use of Pressurised Liquid Extraction resulted in an optimum activity of 339 mg Trolox Equivalents/100 g DW potato peel at 70% ethanol and 125 °C. Therefore the use of Pressurised Liquid Extraction did not enhance extraction in comparison to solid–liquid extracts, but using aqueous ethanol as extraction solvent recovered a higher level of polyphenols than when using 100% methanol.     

Extraction Methods of Citrus Peel Phenolic Compounds

Several extraction methods have been reported in the literature for the extraction of phenols from citrus peel. Extraction methods may cause a degradation of phenolic compounds due to high applied temperature and pressure or long extraction times (conventional solvent extraction, high-pressure extraction). However, other extraction methods are limited by the polarity of phenolic compounds (supercritical CO₂ extraction). Novel techniques of extraction of bioactive compounds have been developed in order to shorten the extraction time, increase the extraction yield, and prevent the degradation of the phenolic compounds. This review provides a critical comparison of the different extraction methods of citrus peel phenolic compounds. The review compiles valuable data that could be useful for the choice of an appropriate extraction method for bioactive compounds from vegetables sources. The main parameters influencing the extraction yield are also discussed.     

Optimization of SC–CO2 extraction of zerumbone from Zingiber zerumbet (L) Smith

Response surface methodology (RSM) was applied to optimize the variables affecting the Supercritical carbon dioxide (SC–CO₂) extraction of non-polar compounds from Zingiber zerumbet using the Box-Behnken design (BBD). Dependent variables were the percentage of the chemical components in the ginger vis a vis α-caryophyllene (y₁), camphene (y₂), and zerumbone (2,6,10-cycloundecatrien-1-one, 2,6,9,9-tetramethyl-) (y₃). Pressure was the most significant parameter affecting the amount of each compound extracted. When temperature was kept constant and pressure was increased, all of the dependent variables increased concomitantly. Since pressure and temperature are two of the major influential factors in the extraction using SC–CO₂, any combination of these two parameters could be selected to ascertain the optimum combination for a particular compound in the extract. Extraction at 30 °C and 55 MPa with total amount of 30 g of CO₂ used was found to maximize all the responses.     

Supercritical CO2 Extraction of Annatto (Bixa orellana) Pigments and Some Characteristics of the Color Extracts

Supercritical CO₂ (SC-CO₂) was used to extract natural food colors from Annatto (Bixa orellana) seeds with different pressures and temperatures. Results from the reverse thin-layer chromatographic separation and GC/MS revealed that SC-CO₂ extracts were composed mainly of bixin, geranylgeraniol and norbixin. Differences in absolute values of solubility parameters (SPD) between these main compounds and SC-CO₂ indicated bixin was more easily extracted than norbixin under conditions used. A higher yield of total pigments was obtained at extraction pressure above 310 bar than at 210 bar and 50°C.     

Protein valorization from ora-pro-nobis leaves by compressed fluids biorefinery extractions

Ora-pro-nobis (Pereskia aculeata) leaves were selected as raw material for biorefinery application based on compressed fluid extractions with green solvents. The fractionation followed the sequential processes: (1) Supercritical fluid extraction (SFE) with CO₂, (2) Gas expanded liquid extraction (GXL), using CO₂:Ethanol, and pressurized liquid extraction (PLE) with ethanol, and (3) Subcritical water extraction (SWE). In vitro analysis of total phenolic and total carotenoid contents and antioxidant activity (DPPH and ORAC) were assessed, stipulating PLE method as the second step of the sequential extractions. Total protein and carbohydrate contents were evaluated from SWE samples. Phenolics profile from PLE and SWE samples was assessed by LC-DAD-ESI-MS/MS, identifying 8 phenolic compounds, with quercetin, isorhamnetin and kaempferol as the main detected flavonoids. SWE at 150 and 185 °C provided the highest protein recovery, as the third step in a sequential process.     

Extraction of Volatile Compounds from Aqueous Solution using Micro Bubble, Gaseous, Supercritical and Liquid Carbon Dioxide

When separating volatile compounds from aqueous solution using supercritical carbon dioxide (SC CO₂), methods to bring SC CO₂ into contact with volatile compounds in the solution are very important. An extraction using micro bubble, gaseous, supercritical, and liquid CO₂ generated by a filter nozzle was carried out. Under optimal conditions (20 MPa, 35°C), > 95% of volatile compounds consisting of 6 to 12 carbon atoms could be removed by extraction for 40 min at CO₂ flow rate, 4.0 g/min. Extraction at 35°C could achieve either a selective or nonselective separation by adjusting the extraction pressure, since the effect of pressure on extraction ratio was most significant at 35°C.     

Chocolate and cocoa: New sources of trans-resveratrol and trans-piceid

trans-Resveratrol and trans-piceid were found for the first time in dark chocolate (at least 0.4 ppm trans-resveratrol and 1 ppm trans-piceid) and cocoa liquor (at least 0.5 ppm trans-resveratrol and 1.2 ppm trans-piceid). Because these compounds are highly sensitive to light, a specific extraction procedure was required to recover them, involving delipidation with toluene and cyclohexane and ethanol/water (80/20, v/v) solid–liquid extraction at 60 °C before reverse-phase HPLC-MS/MS analysis (atmospheric pressure chemical ionization [APCI] in the positive mode). Thanks to an exceptionally high procyanidin content, chocolate products displayed higher antioxidant activity than much more concentrated commercial stilbene extracts.     

Grape marc phenolics: Extraction kinetics,quality and stability of extracts

The solid–liquid extraction of phenolic compounds from different vintage marcs with 60% ethanol was studied and experimental data were elaborated according to the different mathematical models applied in literature to recovery of bioactive constituents from plant materials. A first-order kinetics model was finally selected and the total phenolics diffusivities were calculated (1.23–1.50 × 10⁻¹⁰ m² s⁻¹).     

Supercritical Carbon Dioxide Extraction of Valuable Compounds from Citrus junos Seed

Extraction of Citrus junos seed was carried out at temperatures of 40–70 °C, pressures of 20–50 MPa, and CO₂ flow rate of 3 ml/min with supercritical carbon dioxide to obtain the valuable compounds. Seed oil was also extracted by using Soxhlet extraction with hexane as the solvent during 360 min for comparison with the efficiency of supercritical carbon dioxide extraction. Gas chromatography–mass spectrometry (GC–MS) was used to analyze the components present in the seed oil and Gas chromatography-flame ionization detector (GC-FID) was used to quantify their amounts. Among the conditions studied, the highest extraction yield was obtained at higher pressure and temperature (50 MPa and 70 °C). The extraction yield was about 29.5% of the seed, which was almost comparable to that of hexane Soxhlet extraction (33.8%). The results of the GC–MS analyses showed that the seed oil extracted contained N-methylanthranyl acid methyl, fatty acids (such as palmitic, stearic, oleic, linoleic, and linolenic acid), and physiologically active substances of β-sitosterol and squalene.     

Supercritical carbon dioxide-treated blood orange juice as a new product in the fresh fruit juice market

The work described here deals with the effectiveness of using high-pressure carbon dioxide treatment (HPCD) to stabilise freshly squeezed blood orange juice. Technical planning of a continuous high-pressure supercritical carbon dioxide pilot system, suitable for development on an industrial scale, was carried out in our lab. To determine the optimal operating conditions (temperature, pressure, and CO₂/juice ratio), three different experimental trials were carried out. The first trial was conducted at 230 bar, 36 ± 1 °C, 5.08 L/h juice flow rate, and 3.91 L/h CO₂ flow rate, corresponding to a g_CO2/g_juice ratio of 0.770. The second trial utilised the same conditions except that the operative pressure was reduced (130 bar). The third trial was carried out at 130 bar, 36 ± 1 °C, 5.08 L/h juice flow rate, 1.96 L/h CO₂ flow rate, corresponding to a 0.385 g_CO2/g_juice ratio. The effects of processing were evaluated by determining physicochemical, antioxidant, and microbiological parameters of the treated juices. In addition, once the best operative parameters had been determined, physicochemical, antioxidant, microbiological and sensory evaluation of fresh blood orange juice stabilised by HPCD treatment was carried out during refrigerated storage of juices at 4 ± 1 °C for thirty days. The results showed that HPCD treatment cannot be considered as an alternative to traditional thermal methods but as a new mild technology for producing a stabilised blood orange juice with a shelf-life of 20 days.Industrial relevanceBlood oranges are the main cultivated varieties of Citrus sinensis (L.) Osbeck in Italy. Freshly squeezed blood orange juice exert a high antiradical and antioxidant activity, due to its rich phenolic profile, but its preservation is usually assured by thermal treatment which affects its nutritional and sensory value. In this study we proposed a “milder” continuous HPCD process suitable for implementation on an industrial scale. The HPCD stabilised juice retains its physicochemical, antioxidant, and sensory properties and could be placed within a new retail framework, namely, that of fresh juices with a shelf-life of 20 days.     

Optimisation of mild-acidic protein extraction from defatted sunflower (Helianthus annuus L.) meal

For protein isolation from defatted sunflower meal, mild-acidic extraction was investigated to minimise concomitant oxidation and polymerisation of phenolic compounds and their irreversible binding to proteins. Because of the impaired solubility of sunflower proteins at low pH, the potential of sodium chloride (NaCl) to improve protein extractability was firstly screened for pH 2–11. Increasing NaCl concentrations of the aqueous solvent (c_NaCl) up to 2.8 mol/L enhanced the relative protein yield to almost 80% at ambient temperature and pH 5.6–7.4. As to improved protein recovery at minimal interactions with phenolic acids, the concerted effects of pH (3.2–7.4), c_NaCl (1–3 mol/L), temperature (T, 15–45 °C), and meal-to-solvent ratio (MSR, 0.03 and 0.05 g/mL) on the protein concentration of the extract (c_PE) and the relative protein yield (RPY) were examined, using response surface methodology (RSM). Aside from the prevailing influence of pH value and salt concentration, elevated temperature slightly enhanced protein extraction, whereas MSR mainly influenced c_PE, but hardly RPY. Calculated models proved suitable for the evaluation of extraction processes and the prediction of optimum conditions in terms of high protein yields at the lowest pH possible. Extraction at pH 6.0 was shown to be an appropriate compromise yielding 76–83% of the meal protein, depending on the constraints given. With elevated NaCl concentrations compensating for unfavourable pH conditions, mild-acidic extraction was found to be suitable for the recovery of high-quality sunflower protein in terms of light-coloured protein isolates.     

Characterisation of oleoresins extracted from tomato waste by liquid and supercritical carbon dioxide

Tomato waste is characterised by high amounts of bioactive compounds, such as carotenoids, polyphenols and essential fatty acids. In this work, oleoresins were produced from tomato waste (skins + seeds) and skins using carbon dioxide under liquid and supercritical conditions. The use of ethanol as a co-solvent was also evaluated. Compositions of the oleoresins were analysed and compared to oleoresin obtained by means of an organic solvent, which was used as the control. Oleoresin extraction from the waste by liquid and supercritical CO₂ provided the highest yields (12.5%–12.9%). The extracts obtained from the skins with supercritical CO₂ without ethanol had the highest content of α- and γ-tocopherols (80 and 575 mg/100 g oleoresin, respectively), while lycopene and β-carotene were found at 205 and 75 mg/100 g oleoresin, respectively. The highest quantity of polyphenols (9305 mg GAE/100 g oleoresin) was found in the oleoresin extracted from skin by supercritical CO₂ with ethanol. All oleoresins were characterised by a high antioxidant activity, which improved with the synergy of carotenoids, tocopherols and polyphenols.     

Supercritical CO2 extraction of trans-lycopene from Portuguese tomato industrial waste

Supercritical CO₂ extraction of trans-lycopene from Portuguese tomato industrial wastes (skins and seeds) was carried out in a flow apparatus. The effects of moisture content, feed initial composition, particle size, solvent flow-rate, pressure and temperature on the extraction yield and recovery were evaluated.     

The contribution of high pressure carbon dioxide in the inactivation kinetics and structural alteration of myrosinase

High pressure carbon dioxide (HPCD) has been verified to be an efficient way of inactivating enzyme activity. This work investigates the influence of temperature (T), pressure (P), exposure times (t) on the activity of commercial myrosinase (MYR) submitted to HPCD. Results showed that only 1.00% of MYR activity retained at 22 MPa and 65 °C for 5 min. Moreover, the first‐order reaction kinetic data of MYR inactivation as influenced by pressure of HPCD were analysed. With the pressure rising from 8 to 22 MPa at 55 °C, the inactivation rate constant (k) increased from 0.015 to 0.024 min^?1, while the decimal reduction time (D) decreased from 157.2 to 96.1 min. Additionally, a series of exploratory experiments were conducted to investigate the contribution of the HPCD parameters (T, P and CO₂ dissolution), with analysing circular dichroism spectroscopy and tryptophan fluorescence spectra, illustrate that CO₂ dissolution plays a dominant role in MYR inactivation and structural alteration.