Combination of supercritical fluid extraction with high-speed countercurrent chromatography for extraction and isolation of ethyl p-methoxycinnamate and ethyl cinnamate from Kaempferia galanga L.

Supercritical fluid extraction (SFE) with high-speed countercurrent chromatography (HSCCC) was successfully used for the extraction and isolation of ethyl p-methoxycinnamate (EPMC) and ethyl cinnamate (EC) from Kaempferia galanga L. The SFE parameters including extraction temperature, extraction pressure and entrainer volume were optimized by central composite design (CCD). Then the crude extract was separated by HSCCC with a two-phase solvent system composed of n-hexane:ethyl acetate:methanol:water (7:3:8:2, v/v/v/v) in one-step within 60 min. As a result, 13 mg of EPMC and 2 mg of EC were isolated from 100 mg of crude extract with purities of 98.4% and 98.1%, as determined by HPLC. The structural identification was carried out by UV, MS and NMR spectra.     

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.     

Comparison of Supercritical Fluid Extraction and Liquid Solvent Extraction on Antitumor Diterpenoid from Pteris semipinnata L.

Extraction techniques using Supercritical Fluid Extraction (SFE) and Liquid Solvent Extraction (LSE) were evaluated for the extraction of Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5 F), the antitumor diterpenoid from Pteris semipinnata L. The extracts were analyzed by high performance liquid chromatography (HPLC). SFE experiments showed that many factors had a great impact on the yield and purity of the diterpenoid, such as extraction temperature, pressure, fluid flow rate, extraction time, and modifier. For the SFE process, the optimum operation conditions were as follows: extraction temperature of 328.15 K, extraction pressure of 30 MPa, supercritical CO₂ flow rate of 160 kg/h, extraction time of 4 h, and 10% ethanol as the modifier. Under such a condition, the diterpenoid was almost completely extracted from the material and the yield was approximately 0.504 g/kg dry herb by HPLC analysis. The yield was approximately 3 fold higher than that by liquid solvent extraction. The purity of 5F was 5.148 g/kg dried extract with SFE, it was about 9 fold higher than that by LSE. Mass spectrum data indicated there were two correlative compounds, 5F and its derivative with glycose, in both the extracts, and the ratio of the signal strength of 5F and its derivative was about 3:1 in the SFE extract while that ratio was 1:3 in the LSE extract. The results demonstrated that the supercritical fluid extraction was selective, highly efficient, and with less consumption of organic solvents.     

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.     

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.     

Influence of Extraction Method on Yield,Physicochemical Properties and Tocopherol Content of Manketti (<Emphasis Type="Italic">Schinziophyton rautanenii</Emphasis>) Nut Oil

The influence of extraction method on yield, physicochemical characteristics and tocopherol content of manketti nut oil extracted by four different methods has been determined. Soxhlet (SE) and supercritical fluid (SFE) extractions yielded 45.3 and 44.8%, respectively, while screw press and mechanical shaking extractions had 39.7 and 27.3%, respectively. SPE and SE extractions gave oils that had lower values of unsaponifiable matter (0.70; 0.74%) indicating lower amounts of minor components such as tocopherols (233.13; 290.68 µg/g oil), a greater extent of lipid peroxidation parameters; peroxide values (6.25; 3.01 mequiv O₂/kg), para‐anisidine values (10.22; 9.94), totox value (22.72; 15.96), flavour score (?0.25; 2.11), and high acid values (1.23; 1.03 mg KOH/g oil), respectively, compared to SFE and MSE oils. This was attributed to the high processing temperatures of SPE and SE extractions compared to SFE and MSE oils. Refractive indices (1.485–1.487), iodine values (127.97–129.07, Wijs) and density (0.908–0.914 g/cm³) were not affected by extraction method indicating that the oils generally had the same double bond content. Saponification values (182.98–192.95 mg KOH/g oil) and ester values (181.95–192.11), were not affected by extraction method except for SE oil which had lower values that were speculated to be due to co‐extraction with colour pigments.     

A sequential separation of linderane and norisoboldine using supercritical fluid and ionic liquid-based ultrasonic-assisted extraction from Lindera aggregate

ABSTRACT

Linderane (LDR) and norisoboldine (NOR) are two typical active compounds in Lindera aggregate (Sims.) Kosterm. In this study, a new method of sequential extraction of LDR and NOR from L. aggregate was developed by supercritical fluid extraction (SFE) and ionic liquid-based ultrasonic-assisted extraction (IL-UAE) for the first time. The suitable SFE with CO₂ conditions for LDR were 60 min dynamic extraction time, 40°C temperature and 30 MPa pressure. And the optimal IL-UAE factors for NOR were 2.06 mol/L 1-butyl-3-methylimidazolium bromide ([Bmim] Br) aqueous solution, 44 mL/g liquid–solid ratio, and 67 min ultrasonic time. Compared with the traditional extraction process, the sequential methods not only can obtain higher extraction efficiency, but also can realize the selective extraction for two different kinds of constituents with less consumption of traditional organic solvent. In addition, this environmentally friendly method could be used in a large-scale industry.     

Separation and Purification of Arctiin,Arctigenin, Matairesinol,and Lappaol F from Fructus Arctii by High-Speed Counter-Current Chromatography

Arctiin (I), arctigenin (II), matairesinol (III), and lappaol F (IV) were isolated and purified from the traditional Chinese medicine Fructus Arctii by high-speed counter-current chromatography (HSCCC). The crude extracts from Fructus Arctii were treated with D101 macroporous resin first and divided into two parts: fraction 1 and fraction 2. Fraction 1 was separated by ethyl acetate-n-butanol-water (4:0.5:5, v/v/v) and yielded 164 mg of I from 250 mg of fraction 1. Fraction 2 was separated by n-hexane-ethyl acetate-methanol-water (2:3:2:3, v/v/v/v) and yielded 27 mg of II, 5 mg of III, and 3 mg of IV from 150 mg of fraction 2. The purities of the four compounds were 99.64%, 98.48%, 96.16%, and 91.41%, respectively, as determined by HPLC-DAD. The chemical structures of the isolated compounds were identified by MS, UV, ¹H NMR, and ¹³C NMR analysis.     

Isolation and Purification of Kaempferol-3,7-O-α-L-Dirhamnopyranoside from Siraitia grosvenori Leaves by High-Speed Counter-Current Chromatograph and Its Free Radical Scavenging Activity

Semi-preparative high-speed counter-current chromatography (HSCCC) was successfully used for the isolation and purification of flavonoid glycoside from the leaves of Siraitia grosvenori by using a two-phase-solvent system composed of ethyl acetate–n-butanol–water (4:1:5, v/v/v). kaempferol-3,7-O-α-L-dirhamnopyranoside was obtained in one-step separation and less than 5.5 h from 90 mg of crude extract from the S. grosvenori leaves. The chemical structure of this compound was identified by MS, ¹H NMR, and ¹³C NMR. Free radical scavenging activity of kaempferol-3,7-O-α-L-dirhamnopyranoside was also evaluated and the results showed that it had good free radical scavenging activity with its IC₅₀ value being 3.97 mg/ml.     

Enrichment and Separation of Sinomenine and Acutumine from Sinomenium acutum by pH-Zone-Refining Counter-Current Chromatography

Enrichment and separation of alkaloids from a chloroform extract of Sinomenium acutum has been successfully performed for the first time using pH-zone-refining counter-current chromatography. The two-phase solvent system used for enrichment was composed of Methyl tert-butyl ether (MtBE)–acetonitrile (CH₃CN)–water (4:1:5, v/v), where 10 mM triethylamine (TEA) was added to the upper organic stationary phase as a retainer and 10 mM hydrochloric acid (HCl) to the aqueous mobile phase as an eluter, which could enrich the alkaloids from the crude extract well. For the preparative separation, the solvent system consisted of MtBE–CH₃CN–water (4:0.5:5, v/v) with 10 mM TEA in organic stationary phase and 5 mM HCl in the aqueous mobile phase, which could separate and purify the enriched crude alkaloids successfully. 0.82 g of crude alkaloids was enriched from 1.60 g of chloroform extract in the first step separation. From the enriched crude alkaloids, 376 mg of sinomenine and 85 mg of acutumine were obtained in the second step separation with the purity of 98.1% and 98.7%, respectively. The chemical structures of the isolated compounds were identified by UV, ESI-MS and ¹H NMR.     

An Efficient Strategy Based on Macroporous Resins and Semi-Preparative High Performance Liquid Chromatography for Rapid Separation of Five Flavonoids Components From Flaveria Bidentis(L.) Kuntze

In this study, a simple, rapid, and efficient purification method of five major flavonoids from crude Flaveria bidentis extracts was established. Five flavonoids components were initially obtained by ethanol-water extraction of Flaveria bidentis (L.) Kuntze, followed by using D4020 resin-based column chromatography and semi-preparative high performance liquid chromatography. 9.3 mg hyperoside, 6.5 mg patuletin-3-O-glucoside, 1.7 mg isorhamnetin 3-sulphate, 8.4 mg astragalin, and 4.9 mg 6-methoxykaempferol-3-O-galactoside at high purity of over 94% were obtained from 580 mg Flaveria bidentis extracts. This method is more efficient than HSCCC comparing with the results of these two methods regarding analysts’ purity, recovery, and running time.     

Extraction and Enrichment of Pesticides for Analysis using Binary Supercritical Fluid Mixtures

Abstract

To improve the extraction selectivity in analytical supercritical fluid extraction (SFE), binary mixtures of supercritical fluids have been used to selectively produce lipid‐free extracts of trace pesticide residues for chromatographic analysis. Mixtures of nitrogen and HC‐134a have been used in conjunction with carbon dioxide to selectively extract both fortified and incurred residues from a variety of fatty foods using both experimental and commercially‐available extraction instrumentation. Extraction of fortified pesticides in butter fat showed high recoveries for both organo‐chlorine and ‐phosphorus pesticides with recoveries declining for specific pesticide moieties at the 1 ppb spiking level for CO₂/N₂ mixtures. For CO₂ and HC‐134a mixtures, there was poor and little improvement in analyte recovery relative to using neat SC‐CO₂ for fortified organochlorine pesticides in butter fat at the 0.5–5.0 ppm level. However using a CO₂/HC‐134a mixture for organophosphorus pesticides in butter fat improved recoveries (>90%) of the analytes at the 1–3.5 ppm level relative to using neat SC‐CO₂. Excellent recoveries were obtained for incurred organochlorine and ‐phosphorus pesticides from a variety of food products at ppb levels using either CO₂/N₂ or CO₂/HC‐134 mixtures. Results from these and additional experiments suggest that binary fluid mixtures can significantly reduce the need for additional sample cleanup prior to chromatographic analysis, allowing in some cases, direct injection of the extract after dilution into the gas chromatograph.     

Study on Oleuropein Extraction from Olive Tree (Olea europaea) Leaves by means of SFE: Comparison of Water and Ethanol as Co-Solvent

The present study focuses on developing methods for olive leaf extraction and deals with obtaining extract, rich in oleuropein, which is the most abundant phenolic compound in olive leaves. Supercritical fluid extraction (SFE) was applied to the dried and ground olive leaves by using CO₂ as supercritical (SC) fluid in the presence of water and ethanol as co-solvent. The influences of operating parameters by means of co-solvent content (0-1 mL/min), temperature (50 and 100°C) and pressure (100-300 bar) on both extract and oleuropein yields were investigated. Quantitative analysis was performed by using a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) technique. The experimental results obtained by using SC-CO₂ alone were not satisfactory, and it was seen that addition of a polar modifier is necessary in order to improve yield and selectivity of the process. It was observed that CO₂ modified by water and ethanol showed nearly the same extract performance, where CO₂ modified by water is better for high oleuropein yield.     

Separation and Purification of Bioactive Flavonol Glycosides from Hedyotis diffusa Willd by High-Speed Counter-Current Chromatography

Three flavonoid glycosides including quercetin-3-O-[2″-O-(6″′-O-E-sinapoyl)-β-D-glucopyranosyl]-β-D-glucopyranoside(I), quercetin-3-O-[2″-O-(6″′-O-E-feruloyl)-β-D-glucopyranosyl]-β-D-glucopyranoside(II) and quercetin-3-O-[2″-O-(6″′-O-E-feruloyl)-β-D-glucopyranosyl]-β-D-galactopyranoside(III) were isolated and purified from Hedyotis diffusa Willd by high-speed counter-current chromatography (HSCCC). This run was carried out with a two-phase solvent system composed of n-hexane–ethyl acetate–n-butanol–methanol–1.0% acetic acid (1:1:3.5:1:4.5, v/v) by eluting the lower phase as the mobile phase with a flow-rate at 2.0 ml/min. Consequently, 29.6 mg of I, 35.1 mg of II, 41.3 mg of III with purities of over 95% were obtained from 200 mg of the crude extracts in a single run in less than 130 min. The structure of the isolated compounds was confirmed by MS, ¹H NMR, and ¹³C NMR analysis.     

Response Surface Optimized Ultrasonic-Assisted Extraction of Quercetin and Isolation of Phenolic Compounds From Hypericum perforatum L. by Column Chromatography

The effects of ultrasonic-assisted extraction factors for the main phenolic compound (quercetin) from Hypericum perforatum L. were optimized using the Box–Behnken design (BBD) combined with response surface methodology. The BBD was employed to evaluate the effects of extraction temperature (30–70°C), extraction time (20–80 min), methanol concentration (20–80%, v/v), and HCl concentration (0.8–2.0 M) on the content of one of the major phenolic compounds of quercetin. The extracts were analyzed by high performance liquid chromatography (HPLC). The major phenolic compounds of H. perforatum were isolated and the antioxidant capacity and total phenol content were determined in crude extract and fractions. The optimum conditions were determined as extraction temperature 67°C, extraction time 67 min, methanol concentration 77% (v/v), and HCl concentration 1.2 M. The predicted content of quercetin was 10.81 mg/g dried plant under the optimal conditions and the subsequent verification experiment with 11.09 mg/g dried plant confirmed the validity of the predicted model. The isolated compounds were identified as quercetin, cyanidin, protocatechuic acid, and kaempferol.     

Gurum Seeds: A Potential Source of Edible Oil

Cucurbitaceae family seeds are mostly discarded as agro-industrial wastes. Gurum (Citrullus lanatus var. colocynthoide) is an underutilized wild cucurbit plant, closely related to desert watermelon, which is grown abundantly in some African countries. Gurum seeds can play a significant role in health and nutrition due to their high oil content. This review describes the nutritional composition of gurum seeds and their oil profile. Gurum seeds are a good source of oil (27–35.5%), fiber (26–31%), crude protein (15–18%), and carbohydrates (14–17%). Gurum seeds oil is extracted by supercritical CO₂ (SFE), screw press, and solvent extraction techniques. The gurum seeds oil is composed of unsaturated fatty acids with a high proportion of linoleic acid (C18:2) and oleic acid (C18:1). Gurum seeds oil contains various bioactive compounds, such as tocopherols, phytosterols, and polyphenols. It is reported that solvent extraction gives a higher yield than the screw press and SFE, but the SFE is preferred due to safety issues. More studies are required for producing better quality gurum seeds oil by using novel extraction techniques that can increase oil yield.     

Extraction of sage (Salvia officinalis L.) by supercritical CO2: Kinetic data, chemical composition and selectivity of diterpenes

The supercritical carbon dioxide (SFE) extraction of Dalmatian sage (Salvia officinalis L.) was investigated and compared to extraction performed by Soxhlet ethanol-water (70:30) mixture extraction (SE) and hydrodistillation (HD). The supercritical extraction allowed isolation of wide spectrum of phytochemicals, while other applied methods were limited to either volatiles (HD) or high molecular compounds isolation (SE). The kinetics of the supercritical extraction and fractionation within the pressure range of 10-30 MPa at 50 °C were also analyzed as well as the chemical compositions of total extract and partial or differential fractions isolated at different CO₂ consumption. Volatile fraction could be isolated at low pressure and low CO₂ consumption, whereby the pressures between 10 and 15 MPa followed by increased CO₂ consumption were favourable for obtaining desired selectivity of diterpenes which contain compounds with expressed antioxidative characteristics.     

Supercritical Fluid Extraction and Chromatography of Lipids in Bilberry

A supercritical fluid extraction (SFE) method has been developed for the extraction of lipids in bilberry. Experimental design was used to optimize pressure, temperature and extraction time using CO₂ as solvent. Best SFE condition for total lipids was 450 bar, 60 °C and 45 min. The SFE method was compared to conventional Bligh & Dyer (B&D) extraction. The amount of fatty acid methyl esters (FAME) was found to be 4.84 ± 0.06 mg and 4.564 ± 0.003 mg per g of the freeze‐dried bilberry sample for the developed SFE and B&D methods, respectively, while the amount of total lipids was found to be 54.40 ± 6.06 mg and 65.70 ± 0.67 mg per g of sample for SFE and B&D, respectively. This discrepancy between FAME and total lipids could be explained by the presence of wax esters, sterol esters, carotenoids and phospholipids, as determined by supercritical fluid chromatography.     

Obtaining phenolic compounds from jatoba (Hymenaea courbaril L.) bark by supercritical fluid extraction

The extraction of polyphenol compounds from jatoba (Hymenaea courbaril L. var stilbocarpa) bark using supercritical fluid extraction (SFE) with CO₂ and cosolvents has been investigated. Among the solvent systems studied, SFE using CO₂ and water (9:1, v/v), at 323 K and 35 MPa, presented the best results, with extract yield of 24%, and with high antioxidant activity (IC₅₀ of 0.2 mg/cm³). This solvent system was used to determine global yield isotherms, which were built at 323 and 333 K, and 15, 25, and 35 MPa, using a second lot of jatoba. The highest yield was 11.5% at 15 MPa and 323 K, with maximum total phenolic compounds (TPC) of 335.00 mg TAE/g extract (d.b.) and total tannins content of 1.8 g/100 g raw material. A kinetic experiment was performed using optimized conditions, yielding 18% extract, and the kinetic parameters were used to scale-up the process from laboratory to pilot scale. Chemical analyses showed high content of phenolic compounds in the extracts of jatoba bark mostly due to the presence of procyanidins.     

Supercritical fluid extraction for quality control in beer industry

The knowledge of lipid composition in beer ingredients (malt and corn grits) and wort enables the quality control for final product. Since supercritical fluid extraction (SFE) is an efficient technique for preparing samples for analysis without the use of solvents, in this research Supercritical CO₂ (SC–CO₂) extraction was compared with the traditional Soxhlet one for a gravimetric determination of total lipids on malt and corn grits. The obtained extracts were then analyzed by HPLC-ELSD after TLC separation of triacylglycerols (TAGs) for lipids fingerprint. The extraction of total fats achieved by a 60-min run with pure CO₂ at 65 MPa and 100 °C was 43% higher than that produced by Soxhlet performed for 9 h for malt. The extraction was intermediate for SFE at 60 and 80 °C. The recovery of the TAG obtained with SC–CO₂ at 100 °C was statistically comparable with results from Soxhlet extraction.