Supercritical fluid extraction of naringin from the peel of Citrus paradisi

The highest yield (14.4 g/kg) of naringin, the major flavonoid from the peel of Citrus paradisi L., that could be achieved by supercritical fluid extraction was obtained using supercritical carbon dioxide modified with 15% ethanol and fresh (rather than dried) peels at 95 bar and 58.6 degrees C. This yield is higher than that attained by the conventional technique of maceration, and close to those obtained by reflux and Soxhlet methods. Furthermore, supercritical fluid extraction consumes less solvent and provides a shorter extraction time than conventional extraction methods.     

Combining supercritical fluid extraction of soil herbicides with enzyme immunoassay analysis

Supercritical fluid extraction (SFE) of soil herbicides followed by enzyme immunoassay analysis (EIA) is explained in a step-by-step process. Extracted herbicides, include 2,4-D, simazine, atrazine, and alachlor. The herbicide, trifluralin was not successfully analyzed by EIA because of crossreacting metabolites. Problems with SFE, including uneven packing of cells, leaks, uneven flow and clogging, can largely be eliminated as the method parameters are optimized. It was necessary to add modifiers including methanol or acetone to the SF CO₂ to increase the solubility of the analytes. Detection limits of 2.5 ng/g soil for atrazine and alachlor and 15 ng/g soil for simazine and 2,4-D without concentration of the sample were achieved. Recoveries above 80% and relative standard deviations (RSDs) less than 15% for 2,4-D simazine, atrazine and alachlor were achieved. Atrazine and alachlor recoveries were above 90% with RSDs below 10%. Forty soil samples could be extracted and analyzed in an 8-h day.     

Alpinia oxyphylla Miq. bioactive extracts from supercritical fluid carbon dioxide extraction

In this study, supercritical fluid carbon dioxide extraction technology was developed to gain the active components from a native plant, Alpinia oxyphylla Miq. We studied the biological effects of A. oxyphylla extracts via multiple assays and demonstrated bio-functions at various concentration ranges. Investigations of A. oxyphylla extracts indicated that anti-oxidative properties in dose-dependant manners on radical scavenging activities, reducing power and metal chelating power. The cultured human normal peripheral blood mononuclear cells (PBMCs) were used to test the properties of the extracts in inflammatory cytokine release, and the data did not induce inflammatory molecule releases from un-stimulated PBMCs. A. oxyphylla extracts were able to induce Th1 cytokine IFN-γ released, but not Th2 cytokine IL-13, showing an enhanced anti-bacterial/viral immune function without possible allergic response mediated by IL-13. The extracts also had in vitro mushroom tyrosinase inhibition and cellular tyrosinase melanin decreasing activities in B16F10 cells. In addition, the cell proliferation assay illustrated anti-growth and anti-migration effects in dose-dependent manners of the extracts on human skin melanoma cells, A375 and A375.S2, indicating that the extracts exerted the anti-cancer properties. To our knowledge, this was the first report presenting these bioactivities on A. oxyphylla extracts including antioxidant, anti-inflammation, de-pigmentation and anti-melanoma     

Rapid techniques for the extraction of vitamin E isomers from Amaranthus caudatus seeds: ultrasonic and supercritical fluid extraction

Supercritical fluid extraction (SFE) of seeds of Amaranthus caudatus (Amaranthaceae) and the use of ultrasound as a co-adjuvant in the extraction process were compared with methods traditionally used in the extraction of tocopherols and fatty acids. The use of readily available ultrasound equipment as an adjunct to the classical methods employed for the extraction of tocols provided qualitatively acceptable results more rapidly and more economically. SFE gave quantitatively better yields in shorter times, with solvent-free extracts obtained under conditions that minimised the degradation of thermolabile components. No significant variations were observed in the profile of the fatty acids extracted from amaranth oil by SFE or other methods, thus confirming the qualitative comparability of the faster supercritical extraction with the more time-consuming classical techniques even when processed with the aid of ultrasound.     

Optimisation of supercritical fluid extraction of indole alkaloids from Catharanthus roseus using experimental design methodology--comparison with other extraction techniques

Response surface modelling, using MODDE 6 software for Design of Experiments and Optimisation, was applied to optimise supercritical fluid extraction (SFE) conditions for the extraction of indole alkaloids from the dried leaves of Catharanthus roseus. The effects of pressure (200-400 bar), temperature (40-80 degrees C), modifier concentration (2.2-6.6 vol%) and dynamic extraction time (20-60 min) on the yield of alkaloids were evaluated. The extracts were analysed by high-performance liquid chromatography and the analytes were identified using ion trap-electrospray ionisation-mass spectrometry. The method was linear for alkaloid concentration in the range 0.18-31 microg/mL. The limits of detection and quantification for catharanthine, vindoline, vinblastine and vincristine were 0.2, 0.15, 0.1 and 0.08 microg/mL and 2.7, 2.0, 1.3 and 1.1 microg/g, respectively. The dry weight content of major alkaloids in the plants were compared using different extraction methods, i.e. SFE, Soxhlet extraction, solid-liquid extraction with sonication and hot water extraction at various temperatures. The extraction techniques were also compared in terms of reproducibility, selectivity and analyte recoveries. Relative standard deviations for the major alkaloids varied from 4.1 to 17.5% in different extraction methods. The best recoveries (100%) for catharanthine were obtained by SFE at 250 bar and 80 degrees C using 6.6 vol% methanol as modifier for 40 min, for vindoline by Soxhlet extraction using dichloromethane in a reflux for 16 h, and for 3',4'-anhydrovinblastine by solid-liquid extraction using a solution of 0.5 m sulphuric acid and methanol (3:1 v/v) in an ultrasonic bath for 3 h.     

超临界CO_2萃取冬虫夏草子座挥发性成分的GC-MS研究

报道冬虫夏草挥发性成分的组成,为其进一步的研究工作奠定基础。采用超临界CO2萃取法从冬虫夏草子座中提取挥发性成分,气相色谱-质谱联用技术对其化学成分进行分析。超临界流体萃取物共鉴定了39种组分,占总馏出组分的86.6%以上,占色谱总馏出峰面积的98.56%以上。已鉴定组分中,含量最高的为油酸,相对含量25.6%;其次为亚油酸,相对含量22.67%;再次为棕榈酸11.86%。超临界CO2萃取法能更真实、全面的反映药材中的化学成分,适合于珍稀中药材相关组分的测定。     

Quantification of seed oil from species with varying oil content using supercritical fluid extraction

Introduction: The quantity and composition of seed oil affects seed viability and storability and hence the value of a species as a resource for nutrition and plant conservation. Supercritical fluid extraction with carbon dioxide (SFE‐CO₂) offers a rapid, environmentally friendly alternative to traditional solvent extraction. Objective: To develop a method using SFE‐CO₂ to quantify the seed oil content in a broad range of species with high to low oil contents. Methodology: Seed oil was extracted using SFE‐CO₂ from four crop species representing high, medium and low oil content: Helianthus annuus, Asteraceae, with ca. 55% oil; Brassica napus, Brassicaceae, with ca. 50% oil; Glycine max, Fabaceae, with ca. 20% oil; and Pisum sativum, Fabaceae, with ca. 2% oil. Extraction pressures of 5000, 6000 and 7500 psi and temperatures of 40, 60 and 80°C were examined and a second step using 15% ethanol as a modifier included. Oil yields were compared with that achieved from Smalley Butt extraction. The optimised SFE‐CO₂ method was validated on six species from taxonomically distant families and with varying oil contents: Swietenia humilis (Meliaceae), Stenocereus thurberi (Cactaceae), Sinapis alba (Brassicaceae), Robinia pseudoacacia (Fabaceae), Poa pratensis (Poaceae) and Trachycarpus fortunei (Arecaceae). Results: The two‐step extraction at 6000 psi and 80°C produced oil yields equivalent to or higher than Smalley Butt extraction for all species, including challenging species from the Brassicaceae family. Conclusion: SFE‐CO₂ enables the rapid analysis of seed oils across a broad range of seed oil contents. Copyright © 2008 John Wiley & Sons, Ltd.     

Mathematical modeling and study of mass transfer parameters in supercritical fluid extraction of fatty acids from Trout powder

Mathematical modeling of fatty acids (FAs) extraction from Trout powder by supercritical carbon dioxide was performed in the present work. Trout powder with its low cost contains high amount of essential FAs and it is commonly available as a proper source of FAs. The effect of process parameters, such as pressure (25, 28, 31, 34 and 37 MPa) and temperature (310, 318 and 326 K) of extraction and void fraction of the bed (0.25, 0.35 and 0.45, v/v) on the yield of FAs extraction was examined in a series of experiments conducted in a laboratory scale apparatus. The results indicated a significant increase of extraction yield with an increase of pressure from 25 to 34 MPa, but working at the higher pressure (37 MPa) caused reduction of the extract. Increasing the temperature higher than 318 K revealed significant reduction of the FAs yield and increasing the bed void fraction from 0.25 to 0.45 showed enhancement of the extraction.

The mathematical model was developed considering diffusion-controlled regime in the particle and film mass transfer resistance around the particle with axial dispersion of the bulk phase at dynamic conditions. Henry law was used to describe the equilibrium state of solid and fluid phases. The proposed mass balance equations were numerically solved using implicit finite difference method and the model parameters were correlated using the experimental results of the outlet FAs concentration in the oil extracted at dynamic conditions. Well-known Nelder–Mead method was applied to estimate the four parameters of the model, namely, mass transfer coefficient (k_f), axial dispersion coefficient (D_ax) in the bulk phase, effective diffusivity (D_eff) into the pores and Henry coefficient (H). In the range of studied conditions, the higher extraction efficiency with higher pressure resulted lower correlated H, although the temperature increasing which showed a retrograde phenomena in the FAs yield, revealed H passing though a minimum.     

Supercritical fluid extraction of lipids from the heterotrophic microalga Crypthecodinium cohnii

Microalgae biomass can be a feasible source of ω‐3 fatty acids due to its stable and reliable composition. In the present study, the Crypthecodinium cohnii growth and docosahexaenoic acid (DHA, 22:6ω3) production in a 100 L glucose‐fed batch fermentation was evaluated. The lipid compounds were extracted by supercritical carbon dioxide (SC‐CO₂) from C. cohnii CCMP 316 biomas, was and their fatty acid composition was analysed. Supercritical fluid extraction runs were performed at temperatures of 313 and 323 K and pressures of 20.0, 25.0 and 30.0 MPa. The optimum extraction conditions were found to be 30.0 MPa and 323 K. Under those conditions, almost 50% of the total oil contained in the raw material was extracted after 3 h and the DHA composition attained 72% w/w of total fatty acids. The high DHA percentage of total fatty acids obtained by SC‐CO₂ suggested that this extraction method may be suitable for the production of C. cohnii value added products directed towards pharmaceutical purposes. Furthermore, the fatty acid composition of the remaining lipid fraction from the residual biomass with lower content in polyunsaturated fatty acids could be adequate for further uses as feedstock for biodiesel, contributing to the economy of the overall process suggesting an integrated biorefinery approach.     

Effects of Drying Rates on the Desiccation Tolerance of Citrus maxima ‘Feizhouyou’ Seeds

The effects of drying rates on the desiccation tolerance of Citrus maxima ‘Feizhouyou’ seeds at different developmental stages were studied in this paper. For seeds harvested at 130 days after anthesis (DAA), 190 DAA, 245 DAA and 275 DAA, slow dried seeds had higher desiccation tolerance than those rapid dried, with difference at significant level (P < 005). However, such improvement was little for seeds harvested at 155 DAA and 220 DAA, indicating that effect of drying rate on desiccation tolerance depends on seed developmental stages. These results accorded with previous reports on orthodox soybean seeds and maize embrys. It was suggested that the effects of drying rate on desiccation tolerance of intermediate Citrus maxima ‘Feizhouyou’ seeds mainly resulted from expression and accumulation of some desiccation related proteins induced by slow drying. On the required genetic basis, desiccation tolerance in seeds can be induced only at suitable seed developmental stages.     

柑橘MYB15基因的克隆与表达分析

采用电子克隆和RT-PCR方法从柚(Citrus maxima(Burm.)Merr.)、枳(Poncirus trifoliata(L.)Raf.)和柠檬(Citrus limon(L.)Burm.f.)实生苗中克隆了3个MYB蛋白基因,分别命名为CmMYB15、PtMYB15和ClMYB15;并用实时定量qRT-PCR技术检测了该基因在脱落酸(ABA)、干旱、低温和高盐胁迫处理下的时空表达。结果显示,CmMYB15、PtMYB15和ClMYB15的cDNA序列全长分别为994、992、988 bp,分别编码267、266、265个氨基酸,且编码的氨基酸序列N端均含有2个串联的不完全重复的MYB DNA-binding结构域,由此推测该3个基因均属于R2R3亚类;MYB15基因均能被ABA、干旱、低温和高盐胁迫诱导表达,且在柚、枳和柠檬中存在表达差异。本研究表明柚CmMYB15、枳PtMYB15和柠檬ClMYB15是MYB基因家族成员,可能在柑橘响应非生物胁迫过程中起到一定的作用。     

Effects of pressure and temperature of supercritical carbon dioxide on the extraction of triacylglycerols from plant tissue

The rate of extraction of triacylglycerols with supercritical carbon dioxide can be greatly enhanced by raising the pressure of the fluid to 600 bar, or higher, and its temperature to 60°C, or higher. Both the amount of carbon dioxide and the time required for complete extraction are reduced at such high pressures and temperatures.     

Extraction conditions affecting supercritical fluid extraction (SFE) of lycopene from watermelon

Lycopene, a carotenoid linked to protection against certain forms of cancer, is found in produce such as papaya, red-fleshed tomatoes, grapefruit and watermelon. The preparation of a supercritical CO2 (SC-CO2) watermelon-lycopene extract could serve as a food grade source of this carotenoid. This study established preliminary conditions for enhancing SC-CO2 extraction of lycopene from watermelon. Freeze-dried watermelon was extracted with SC-CO2 and ethanol as an organic co-solvent. The lycopene concentration was determined by HPLC, with absorbance measured at 503 nm. In an initial set of experiments, the effects of extraction temperature (70-90 degrees C), pressure (20.7-41.4 MPa) and co-solvent ethanol addition (10-15%) were evaluated. A lycopene yield of 38 microg per gram of wet weight was obtained at 70 degrees C, 20.7 MPa, and 15% by volume ethanol. The extraction of fresh (non-freeze-dried) watermelon yielded 103+/-6 microg lycopene per gram fresh fruit weight. Of the parameters tested, temperature had the most effect on lycopene yield. Thus, in another set of experiments, the temperature was varied from 60-75 degrees C at an extraction pressure of 20.7 MPa in the presence of 15% ethanol. Studies showed that freeze-dried watermelon flesh loses lycopene in storage. In accounting for lycopene storage losses, lycopene yields at 60 degrees C extraction temperature were 14% greater than those obtained at 70 degrees C.     

超临界CO2萃取石香薷挥发油工艺的研究

石香薷（Molsa chinensis）为多年生草本植物,具有重要的药用价值。本文首次探讨了超临界CO2萃取的压力,温度,流量,时间等条件对石薷挥发油收率的影响,确定其最佳萃取条件,萃取压力15MPa,温度45℃,CO2流量25kg/h和90min,得率为3.56%,水蒸气蒸馏提取得率为1.62%,石油醚提取得率为1.87%,结果表明超临界CO2萃取挥发油比后二者质优,收率大大提高,萃取时间短,无溶剂残留。     

HPLC法测定不同产地和品种柠檬果皮中柠檬苦素和诺米林含量

该研究建立了同时测定檬苦素和诺米林含量的高效液相色谱法,并采用该方法对8种不同柠檬样品中柠檬苦素和诺米林含量进行了测定。色谱条件如下:安捷伦Eclipse XDB-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为0.1%磷酸水溶液(A)-乙腈(B),梯度洗脱,流速为1.0 mL·min^-1,柱温为30℃,波长为210 nm。结果表明:3个产地尤力克果皮中柠檬苦素含量依次为云南瑞丽(EUR_D)、四川安岳(EUR_C)和广东河源(EUR_Y),诺米林含量依次为EUR_D、EUR_C和EUR_Y;不同品种柠檬果皮中柠檬苦素含量依次为北京柠檬(M_NM)、粗柠檬(R_NM)、无核柠檬(S_NM)、热那亚(RNY)、里斯本(LSB),诺米林含量依次为S_NM、R_NM、LSB、RNY和M_NM。综上结果表明,不同产地和品种柠...     

Selective separation of cis-trans geometrical isomers of beta-carotene via CO2 supercritical fluid extraction

We investigated a novel method for the selective separation of beta-carotene isomers from a freeze-dried powder of the algae Dunaliella bardawil using supercritical fluid extraction. The separation method relies on the different dissolution rate of the 9Z and all-E isomers of beta-carotene in SC-CO(2). At first, the equilibrium solubility of the two isomers in SC-CO(2) was determined at the extraction conditions of 44.8 MPa and 40 degrees C. The solubility of the 9Z isomer was found to be nearly 4 times higher than that of the all-E isomer (1.92 x 10(-5) g all-E isomer/g CO(2) compared to 7.64 x 10(-5) g 9Z isomer/g CO(2)). When supercritical fluid extraction was applied to a carotenoid concentrate from the algae (29 wt% beta-carotene) or a freeze-dried powder of the algae (3.1% beta-carotene), a selective separation of the 9Z/all-E isomers of beta-carotene was obtained. Thirty-nine percent recovery of beta-carotene with 80% purity of 9Z isomer was achieved at the initial stages of extraction (40 mL CO(2)). The extraction rate of beta-carotene from the freeze-dried algae powder was slower than that from the carotenoid concentrate, resulting in a reduction in the recovery and purity of the 9Z isomer. This indicates that even at the initial stage of the extraction the internal mass resistance is significant. Isomer purity and recovery could be enhanced upon grinding of the algae powder.     

Application of mixtures of tartaric acid derivatives in resolution via supercritical fluid extraction

Racemic N-methylamphetamine (rac-MA) was resolved with 2R,3R-tartaric acid (TA) and its derivatives (O,O'-dibenzoyl-(2R,3R)-tartaric acid monohydrate (DBTA) and O,O'-di-p-toluoyl-(2R,3R)-tartaric acid (DPTTA)), individually and using them in different combinations. After partial diastereomeric salt formation, the free enantiomers were extracted by supercritical fluid extraction using carbon dioxide as solvent. DBTA and DPTTA are efficient resolving agents for rac-MA, the best chiral separation being obtained at a molar ratio of 0.25 resolving agent to racemic compound for both resolving agents (ee(E) = 82.5% and ee(E) = 57.9%, respectively). Compared with the two other acids, TA is practically unsuitable for enantiomer separation (ee(E) < 5%). Applying a mixture of one individually active and one ineffective acid in half the equivalent molar ratio, when the acids are in 1:1 ratio in the mixture, the resolution efficiency values obtained exceeded those obtained by using the components individually. Decreasing the molar ratio of resolving agent mixture to 0.25, at which the individual resolving agents give the best chiral separation, the obtained resolution efficiency values did not differ significantly from those expected. The outcome of the resolution process depended only on the amount of the individually active resolving agents in the mixture.     

猴头菌子实体超临界流体技术萃取活性物质初探

以猴头菌子实体为原料,萃取物得率作为指标,采用CO₂超临界流体萃取技术,以萃取压力和CO₂流量等参数为考察因素,结合正交试验获得优化的萃取工艺：萃取压力为30MPa,温度为45℃,时间为1.5h,CO₂流量为20g/min,夹带剂（乙醇）与猴头菌子实体的物料比为5:1（mL:g）,在此条件下,萃取物得率为2.78%。与猴头菌子实体的醇提物相比,猴头菌子实体超临界萃取物具有更好的体外抗氧化能力和抗肿瘤活性,本研究结果为合理地开发和利用猴头菌子实体超临界萃取物产品提供科学的数据。     

Persistence of 14C-gibberellin A3 on the surface of Citrus fruit peel and on inert glass surfaces

The persistence of gibberellin A₃ on plant surfaces was examined using fruit of Marsh seedless grapefruit (Citrus paradisi Macf.) and an inert glass model system. ¹⁴C-gibberellin A₃ was applied to surfaces in aqueous treatment solutions or in waxing solutions. Dried-out treatment residues were removed by washing and analyzed for total and GA₃-like radioactivity. Gibberellin A₃ persisted without significant loss for at least 7 d in aqueous treatment solutions (pH 4.0 or 6.2) but was less persistent in the pH 10.4 waxing solution (t1/2=7 d).Loss of total peel surface radioactivity was fast during the first 3 days, slowing down afterwards. After 14 days 73% of the initial radioactivity could still be recovered from fruit peel surface and 70% of the recovered radioactivity was still in the form of gibberellin A₃. Gibberellin A₃ was somewhat more persistent in residues from pH 4 than pH 7 treatment solutions. Light had a slight enhancing effect on gibberellin A₃ decomposition on fruit peel under growth chamber conditions. After 12 d at 100% relative humidity, 88% of the radioactivity on glass surfaces was still in the form of gibberellin A₃, as against 45% at a relative humidity of 50%. Simulated field conditions, combining daily fluctuations in light, temperature and relative humidity, markedly enhanced gibberellin A₃ decomposition on glass surfaces (t1/2=2 d). Gibberellin A₃ was very persistent (90% after 9 d) in the waxing residues on fruit peel surface.Abbreviations GA₃ gibberellin A₃ - RH relative humidity