Supercritical CO2 extraction of pigment components with pharmaceutical importance from Chlorella vulgaris

BACKGROUND: Chlorella vulgaris is a green microalgae that contains various pigment components of carotenoids and chlorophylls. Supercritical CO₂ is widely used for extraction of pharmaceutical compounds because it is non‐oxic and easily separated from extracted material by simply depressurizing. In this work, pharmaceutical compounds from Chlorella vulgaris have been extracted using supercritical CO₂ with or without entrainer at various extraction conditions. RESULTS: Based on high performance liquid chromatography (HPLC) analysis, the extracts contained pigment components, such as lutein, β‐carotene, chlorophyll a and b. Higher extraction pressure and temperature promoted higher lutein extraction by supercritical CO₂. The optimum pressure and temperature for extraction were obtained as 50 MPa and 80 °C. Ethanol as an entrainer was more effective than acetone for the extraction of pigment components. Pigment components in the extract obtained by supercritical CO₂ with and without entrainer were compared with the extract obtained by a conventional extraction method. CONCLUSION: Supercritical CO₂ has been successfully applied for the extraction of pigment components from Chlorella vulgaris. Supercritical CO₂ enabled high selectivity for lutein extraction; however, the lutein yield was lower than that obtained by extraction using supercritical CO₂ with ethanol and soxhlet. Copyright © 2008 Society of Chemical Industry     

Supercritical CO2 Extraction of Chlorophyll a from Spirulina platensis with a Static Modifier

Supercritical CO₂ extraction with a static modifier was applied to extract chlorophyll a from Spirulina platensis. The effects of the process were investigated by single‐factor and response surface analysis experiments. The optimal process parameters for supercritical CO₂ extraction were determined to be: ethanol/water as the modifier, 40 vol.‐% water content in the modifier, 21.2 mL modifier volume, 1 h static soaking time, 2 h dynamic extraction time, 48.7 MPa extraction pressure, 326.4 K extraction temperature, and 10 g min^–1 CO₂ flow rate. The optimized chlorophyll a extraction yield was 6.84 mg g^–1. A comparison of the experimental results suggested that the yield of chlorophyll a by supercritical CO₂ extraction with modifier was higher than that obtained by conventional solvent extraction.     

Supercritical carbon dioxide essential oil extraction of Lamiaceae family species: Mathematical modelling on the micro-scale and process optimization

In this paper the essential oil supercritical carbon dioxide extraction from leaves of Lamiaceae family species was studied. Recent investigations of Lamiaceae family essential oil storage have shown that most of the oil is found in peltate glandular trichomes on the leaf surface. The effect of supercritical CO₂ on the peltate glands was investigated by Scanning Electron Microscopy. It was observed that exposure to supercritical CO₂ led to disruption of the peltate glands and essential oil release. This phenomenon was used as a basic hypothesis of the mathematical model of the supercritical fluid extraction with CO₂. The model was applied to simulate basil, rosemary, marjoram and pennyroyal supercritical CO₂ extraction on the existing experimental data. An average deviation from the experimental data was less than 0.83%. The model results indicated a possibility of a decrease in the supercritical CO₂ consumption by modified and optimized processing of Lamiaceae family herbaceous material.     

Supercritical carbon dioxide extraction,purification, and characterization of wax from sorghum and sorghum by-products as an alternative natural wax

The objective of this work was to obtain high purity natural wax from sorghum and by-products of sorghum processing (sorghum dried distillers grains with solubles [DDGS] and sorghum bran) using a green process based on supercritical carbon dioxide (SC-CO₂). SC-CO₂ extractions were carried out at varying temperatures (50, 70°C) and pressures (30, 40 MPa) at a CO₂ flow rate 1 L/min for 120 min. Significantly higher wax yield (4.9%) from DDGS was obtained by SC-CO₂ at 40 MPa/70°C compared with whole kernel (0.6%) and bran (3.3%) (p < 0.05). The yield of the extracts obtained by SC-CO₂ extraction was higher than that of the conventional hexane extraction for all three sorghum sources. The highest fraction of wax in the SC-CO₂ extracts was obtained from whole kernel extracts (89%), whereas it was 53.3% from the DDGS and 26.8% from the bran at the same extraction conditions. SC-CO₂ and hexane extracts from sorghum whole kernel shared a similar melting peak temperature of 76.3–77.9 and 79.7°C, respectively, while DDGS and bran extracts by SC-CO₂ showed a much lower melting temperature in the range of 50.7–61.9°C, indicating the presence of lower melting point components such as triacylglycerols. However, the melting points of the DDGS and bran extracts after ethanol purification were significantly increased with the observed peak temperature of 80.8 and 82.0°C, respectively. While the wax yield from DDGS and bran was higher than that of whole kernel, the sorghum whole kernel feedstock was found to be a more feasible feedstock to obtain higher purity wax.     

Supercritical CO2 extraction of accumulated capsidiol from biotic elicitor‐activated Capsicum annuum L fruit tissues

This work investigates the supercritical CO₂ extraction of capsidiol from pepper fruit tissues activated with Alternaria alternate (Fr) Keissler suspension culture as a biotic elicitor. Capsidiol production in the fruit tissue was markedly increased by the treatment with a biotic elicitor and reached its maximum level after 4 days of elicitation. The effects of separation parameters such as temperature, pressure, supercritical solvent flow rate, particle diameter and also initial capsidiol concentration were investigated on solubility, initial extraction rate and extraction yield. The optimal extraction conditions were obtained at the temperature of 40 °C, the pressure of 400 bar, the supercritical CO₂ flow rate of 2 cm³ min^?1, and the average particle diameter of 116 µm. The results showed that the ratio of the supercritical CO₂ extraction yield to the organic solvent extraction yield was changed from 84 to 97 wt‐% depending on the initial capsidiol concentration. Copyright © 2004 Society of Chemical Industry     

Extraction of wheat germ oil by supercritical CO<Subscript>2</Subscript>: Oil and defatted cake characterization

In this paper the working conditions for the extraction of wheat germ oil in a supercritical CO₂ pilot plant of 1-L-extraction capacity were studied. The best conditions were: pressure, 38 MPa; temperature, 55°C; wheat germ particle size, about 0.35 mm; CO₂ flow rate, 1.5 L min⁻¹. These conditions gave yields of about 92% of total oil after 3 h of processing. The obtained oils and the partially defatted cake were investigated with regard to their FA, tocol (tocopherol and tocotrienol), carotenoid, and sterol compositions and to their quality characteristics (FFA, PV, para-anisidine value, and color of the by product). Moreover, the oil quality was evaluated in relation to the progress of the supercritical extraction.     

Effects of basic modifiers on sfe efficiencies of ephedrine derivatives from plant matrix

The effect of basified modifiers in supercritical CO₂ (SCF-CO₂) such as diethylamine has been investigated on the SFE efficiencies of ephedrine derivatives (e.g. methylephedrine, norephedrine, ephedrine, and pseudoephedrine) from aerial parts ofEphedra sinica. The addition of basified modifiers in SCF-CO₂ showed a greater enhancement of SFE efficiency of ephedrine derivatives than pure modifiers. These results might be due to the fact that the salts ofEphedra alkaloids (insoluble in CO₂) in plant tissues would be changed to their free bases (freely soluble in CO₂) by basified modifiers such as diethylamine in methanol. In addition to enhancing the solubilities, it could increase the degree of desorption of the compounds from plant matrix. Also, the SCF-CO₂ modified with methanol basified with diethylamine showed a difference of extractability of ephedrine and its diastereomer, pseudoephedrine. This paper was presented at The 5th International Symposium on Separation Technology-Korea and Japan held at Seoul between August 19 and 21, 1999.     

Supercritical Carbon Dioxide Extraction of <Emphasis Type="Italic">Microula sikkimensis</Emphasis> Seed Oil

The seed oil of Microula sikkimensis had been intensively studied due to its pharmacological actions. In the present study, seed oil of Microula sikkimensis was extracted using supercritical fluid extraction (SFE). Determinations of the extracts composition were performed by gas chromatography (GC). An orthogonal array design (OAD), OA₉ (3⁴), was employed for optimization of the supercritical fluid extraction of the compound with regard to the various parameters. Four factors, namely pressure (21.0–27.0 MPa), the dynamic extraction time, temperature, and CO₂ flow rate of the supercritical fluid, were studied and optimized by a three-level OAD. The effects of the parameters on the yield of seed oil were studied using analysis of variance (ANOVA). The results revealed that the pressure had a significant effect on the yield of seed oil (p < 0.05), while the other three factors, i.e., CO₂ flow rate, dynamic extraction time and temperature, were not identified as significant factors under the selected conditions based on ANOVA. The results show that the best values for the extraction condition of seed oil was pressure 24.0 MPa, extraction time 3 h, temperature 45 °C and a CO₂ flow rate 20 L/h in the 20-L vessel.     

Pilot-scale supercritical carbon dioxide extraction and fractionation of wheat germ oil

There is a need for the development of new processing techniques to facilitate vegetable oil extraction and refining while sustaining the nutritional components naturally present in edible oils and reducing the adverse impact of oil processing on the environment. In this study supercritical carbon dioxide (SC−CO₂) extraction and fractionation techniques were examined as alternative methods to obtain wheat germ oil (WGO) of high quality and purity. It was shown that the SC−CO₂ extraction technique is effective in extraction of WGO. There was no significant difference in the FA composition of SC−CO₂- and hexane-extracted WGO. Both hexane-and SC−CO₂-extracted WGO were rich in α-tocopherol. Moisture content of the SC−CO₂-extracted oil was higher than that of the hexane-extracted oil. Solvent/feed ratio had a significant effect on the SC−CO₂ extraction yields. This study demonstrated that supercritical fluid fractionation was a viable process to remove FFA efficiently from both hexane-and SC−CO₂-extracted WGO while retaining bioactive oil components in the final product.     

Ethanol-Modified Supercritical Carbon Dioxide Extraction of the Bioactive Lipid Components of <Emphasis Type="Italic">Camelina sativa</Emphasis> Seed

Camelina sativa seed is an underutilized oil source that attracts a growing interest, but it requires more research on its composition and processing. Its high omega‐3 content and growing demand for clean food processing technologies make conventional oil extraction less attractive. In this study, the effect of extraction methods on the bioactive lipid composition of the camelina seed lipid was investigated, and its bioactive lipid composition was modified at the extraction stage using ethanol‐modified supercritical carbon dioxide (SC‐CO₂). Ethanol‐modified SC‐CO₂ extractions were carried out at varying temperatures (50 and 70 °C), pressures (35 and 45 MPa), and ethanol concentrations (0–10%, w/w), and were compared to SC‐CO₂, cold press, and hexane extraction. The highest total lipid yield (37.6%) was at 45 MPa/70 °C/10% (w/w) ethanol. Phospholipids and phenolic content increased significantly with ethanol‐modified SC‐CO₂ (p < 0.05). SC‐CO₂ with 10% (w/w) ethanol concentration selectively increased phosphatidylcholine (PC) content. Apparent solubility of camelina seed lipids in SC‐CO₂, determined using the Chrastil model, ranged from 0.0065 kg oil/kg CO₂ (35 MPa/50 °C) to 0.0133 kg oil/kg CO₂ (45 MPa/70 °C). Ethanol‐modified SC‐CO₂ extraction allowed modification of the lipid composition that was not possible with the conventional extraction methods. This is a promising green method for extraction and fractionation of camelina seed lipids to separate and enrich its bioactives.     

Extraction of brown coal with carbon dioxide at supercritical parameters

The composition of extracts from coal obtained by extraction with CO₂ at supercritical parameters was studied. It was shown that CO₂ in a supercritical state possesses high selectivity for predominantly dissolving hydrocarbons from the entire range of bituminous substances and a methane-naphthene fraction from hydrocarbons; this can be used in the complex processing of raw coal.     

Supercritical CO2 extraction of ucuúba (Virola surinamensis) seed oil: global yield,kinetic data,fatty acid profile,and antimicrobial activities

Virola surinamensis is an abundant floodplain tree, popularly known as ucuúba, that grows in the Amazon. In this study, ucuúba seed oil was obtained by supercritical fluid extraction under different operating conditions, as well as Soxhlet extraction. The operating conditions for supercritical extraction were an extraction temperature of 40, 60, or 80?°C, a pressure of 350?bar, and a CO₂ mass flow of 7.9?×?10^?5 kg/s. The supercritical extraction curves were fitted to mass transfer models, and the fatty acid profiles of the extracts were determined by gas chromatography. The antimicrobial activity was assessed against Candida albicans, Staphylococcus aureus, and Escherichia coli. The highest yield obtained using supercritical CO₂ was 64.39% and the lowest was 59.21%. The phytochemical analysis showed the presence of steroids, terpenes, coumarins, and phenolic compounds. All ucuúba oil samples showed antioxidant activity. Regarding the antimicrobial activity, ucuúba oil only showed activity against S. aureus.     

Surface Composition of Myrmecophilic Plants: Cuticular Wax and Glandular Trichomes on Leaves of <Emphasis Type="Italic">Macaranga tanarius</Emphasis>

Primary plant surfaces, covered with cuticles consisting of cutin and waxes, are important substrates for interaction with insects. The composition of leaf surfaces of the myrmecophilic plant Macaranga tanarius was studied. The prenylated flavanone nymphaeol-C was identified in surface extracts and was localized exclusively in glandular trichomes on the abaxial leaf side. The epidermal pavement cells surrounding these trichomes were covered with a smooth film of epicuticular wax from which few small wax crystals protruded. The epicuticular wax amounted to approximately 8 μg cm⁻², corresponding to 85% of the wax load on the adaxial as well as the abaxial leaf sides. The epicuticular wax mixtures from both leaf surfaces contained more than 70% primary alcohols, 14% fatty acids, 2% aldehydes, and traces of alkyl acetates, with chain lengths ranging from C₂₀ to C₃₈. In contrast, the intracuticular wax layer was largely dominated by triterpenoid alcohols α-amyrin, β-amyrin, and lupeol. Consequently, these characteristic compounds are not available for direct contact with insects on the plant surface.     

Applications of supercritical CO2 extraction to microalgae and plants

A flow apparatus was constructed to carry out studies of supercritical fluid extraction in the temperature range 298·15–353·15 K and pressures up to 40 MPa. To test the apparatus, studies on the solubility of naphthalene in CO₂ at 308·15, 313·15 and 318·15 K and pressures up to 35 MPa were carried out. These experimental data were correlated through the Peng–Robinson equation of state. Samples of the microalgae Botryococcus braunii and Chlorella vulgaris which produce diolefines C₂₅–C₃₁ and carotenoids, respectively, were submitted to supercritical CO₂. The extraction yields for these compounds were studied and compared with those obtained using organic solvents. Supercritical CO₂ was also used to produce extracts from rosemary leaves (Rosmarinus officinalis L.), which were compared with those obtained by hexane extraction.     

Antiradical Efficiency of Essential Oils from Plant Seeds Obtained by Supercritical CO2, Soxhlet Extraction,and Hydrodistillation

The effect of supercritical CO₂ (SCCO₂) extraction conditions (pressure and temperature) on the system performance as well as the antiradical efficiencies of the essential oils from Japanese pepper (Xanthoxylum piperitum DC.), cardamom (Elettaria cardamomum Maton), and fennel (Foeniculum vulgare) seeds were investigated. A control study with the conventional Soxhlet extraction and hydrodistillation was also conducted to compare the performance of those processes. Antiradical efficiencies were investigated by utilizing 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay with a UV-vis spectrophotometer. Higher pressure and temperature had positive effects on the supercritical process performance due to higher CO₂ density and substrate solubility in SCCO₂. Antiradical efficiencies of cardamom and pepper were almost the same, being significantly higher than that of fennel seeds. However, this effect decreased dramatically for all the spices when the extraction method was changed to Soxhlet extraction and hydrodistillation. SCCO₂ extraction was found to yield more quality and effective essential oils than Soxhlet extraction and hydrodistillation.     

Removing paraffin‐based wax coatings from old corrugated containers using supercritical carbon dioxide

Supercritical carbon dioxide (SC‐CO₂) extractions of paraffin‐based wax coatings from saturated and curtain‐coated old corrugated containers (OCC) are reported. Extractions were performed in a 500‐mL reactor (300 bar, 100°C, 50 g CO₂/min and 1 h). Wax removal efficiencies of 98 and 70% for saturated and curtain‐coated OCC, respectively, were obtained. Under similar conditions, extractions in the presence of water resulted in an extraction efficiency of 99% for saturated OCC. Decreasing the operating pressure to 200 bar decreased the extraction efficiency to approximately 50%. Gas chromatography (GC) of the wax coatings on OCC, before and after extraction with SC‐CO₂, showed a slight shift in the molecular weight distribution of the paraffin wax (after SC‐CO₂ extraction) toward higher molecular weights for both saturating wax and curtain‐coating wax. There was no evidence of chemical degradation or modification of the paraffin wax coatings by SC‐CO₂. The packing density, packing arrangement, and dimensions of the curtain‐coated OCC in the extraction apparatus affected the extraction efficiency. Loose packing compared to tight packing, 1 × 1 cm squares versus 1 × 20 cm strips, had higher extraction efficiencies; a random packing arrangement was better than packing with the fluting material in the direction of SC‐CO₂ flow. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 83: 2699–2704, 2002     

Leaf Epicuticular Wax Chemicals of the Japanese Knotweed Fallopia japonica as Oviposition Stimulants for Ostrinia latipennis

Extraction, fractionation, and gas chromatography−mass spectrometry analyses guided by bioassays have shown that n-alkanes and free fatty acids in leaf epicuticular wax of the Japanese knotweed Fallopia (Reynoutria) japonica stimulate oviposition in the Far-Eastern knotweed borer, Ostrinia latipennis (Lepidoptera: Crambidae). n-Alkanes made up 48.1% of the total amount of epicuticular wax, and their carbon chain length was in the C₁₆−C₃₃ range, with n-nonacosane (n-C₂₉) most abundant, followed by n-C₂₇, n-C₂₅, and n-C₃₁. Free fatty acids with C₉−C₂₂ accounted for 22.3%, and hexadecanoic acid was predominant. A mixture of authentic n-alkanes and fatty acids of the composition found in the epicuticular wax, a mixture of n-alkanes, and a mixture of fatty acids significantly enhanced oviposition. Thus, it was demonstrated that both n-alkanes and free fatty acids in leaf epicuticular wax of F. japonica are naturally occurring oviposition stimulants for O. latipennis.     

Mathematical Modeling of Ocimum basilicum L. Supercritical CO2 Extraction

The effects of pressure and temperature on supercritical extraction of Ocimum basilicum L. in terms of extraction yield and aromatic compounds were investigated. Linalool, eugenol, and δ‐cadinene were indentified as three main compounds of the prepared extracts. The dominant compound in all investigated supercritical basil extracts was linalool. Within the experimental range the extraction parameters had a significant influence on the extraction yield. The obtained results for the sweet basil total extract, cadinene, and linalool best matched with the Gordillo et al. model, therefore, the solubility of total extract, linalool, and cadinene in supercritical CO₂ at investigated temperatures is described as a function of pressure.     

Supercritical CO2 Extraction of Essential Oil from Origanum Vulgare L.: Experiments and Mathematical Modeling

In this work, the supercritical CO₂ extraction of essential oil from Origanum Vulgare L. was investigated and modeled. An orthogonal test and ANOVA indicated that extraction pressure, extraction temperature, and extraction time had significant influence on extraction effects. Based on experiments, a mathematical model depended on mass conservation equation was established to describe and simulate supercritical CO₂ extraction of essential oil from Origanum Vulgare L. The mean diameter, accumulation properties, and the inside and outside transfer properties of extracted material particles were considered in the model. The model was solved numerically with the finite difference method and Runge-Kutta method synthetically. Model estimation was validated with small scale experimental data. Moreover, the effects of extraction pressure, extraction temperature, extraction time, concentration, and the flow rate of the entrainer on mass of essential oil were investigated using the model.     

Supercritical fluid extraction and bioassay identification of prodrug substances from natural resources

For arbitrarily chosen thirty types of natural resources which have been widely used in oriental traditional herb medicine, supercritical CO₂ extraction (SFE) and organic liquid solvent extraction (LSE) withn-hexane, chloroform and methanol were carried out to extract pharmaceutical substances. To evaluate relative advantages and shortcomings between the SFE and LSE, five types of bioactivity assays as well as gas- and thin layer-chromatographic analysis were performed for all the extracts obtained by the two extraction methods. Types of bioassays performed included cytotoxicity, bleb forming, DNA binding, oxygen free radical scavenger and Xanthine oxidase inhibitor tests. To evaluate economic viability of the SFE over the traditional LSE, extractability of prodrug substances was evaluated as the functions of extraction temperature and pressure. SFE was proven to be a feasible alternative over LSE. Also, the optimum SFE conditions which provided maximum extraction and cytotoxicity for each selected sample were presented.