Effect of extraction method on the yield of furanocoumarins from fruits of Archangelica officinalis Hoffm

Optimal conditions for the extraction and analysis of furanocoumarins from fruits of Archangelica officinalis Hoffm. have been determined. The following extraction methods were used: exhaustive extraction in a Soxhlet apparatus, ultrasonication at 25 and 60 degrees C, microwave-assisted solvent extraction in open and closed systems, and accelerated solvent extraction (ASE). In most cases the yields of furanocoumarins were highest using the ASE method. The effects of extracting solvent, temperature and time of extraction using this method were investigated. The highest yield of furanocoumarins by ASE was obtained with methanol at 100-130 degrees C for 10 min. The extraction yields of furanocoumarins from plant material by ultrasonication at 60 degrees C and microwave-assisted solvent extraction in an open system were comparable to the extraction yields obtained in the time- and solvent-consuming exhaustive process involving the Soxhlet apparatus.     

Induction of secondary metabolite production in transformed callus of Ammi majus L. grown after electromagnetic treatment of the culture medium

High-performance liquid chromatography analysis of crude chloroform and methanol extracts as well as methanol preparations subjected to enzymatic hydrolysis indicated two-fold higher accumulation of umbelliferone in transformed Ammi majus L. callus grown on medium exposed to ADR-4^® (energy stimulator plate) in comparison to the control cultures. Elicitation with ADR-4^® induced also two times higher accumulation of bergapten. Three different techniques: Soxhlet extraction, ultrasonication and accelerated solvent extraction (ASE) applied for the extraction of coumarins, furanocoumarins and corresponding glycosides from transformed callus of A. majus elicited by ADR-4^® plates were compared. ASE is shown to be the most efficient extraction method.     

An ecofriendly approach to process rice bran for high quality rice bran oil using supercritical carbon dioxide for nutraceutical applications

An integrated approach to extraction and refining of RBO using supercritical carbon dioxide (SC-CO2) in order to preserve the nutritionally important phytochemicals is reported here. Process variables such as pressure, temperature, time, solvent flow rate and packing material on extraction yield and quality of RBO were investigated using a pilot model SC-CO2 extraction system. Three isobaric (350, 425 and 500 bar), three isothermal temperatures (50, 60 and 70 degrees C), three extraction times (0.5, 1 and 1.5h), at 40/min CO2 flow rate and three packing materials (pebbles, glass beads and structured SS rings) were employed. The RBO yield with SC-CO2 extraction increased with temperature and time under isobaric conditions. At the 60 degrees C isotherm, an increase in the RBO yield was obtained with an increase in the pressure and time. The RBO yield increased significantly with structured SS rings used as packing material. The RBO extracted with SC-CO2 had negligible phosphatides, wax and prooxidant metals (Fe and Cu) and was far superior in color quality when compared with RBO extracted with hexane. At the optimum condition of extraction at 500 bar, 60 degrees C for 1.5h, with structured SS rings used as packing material, the yield of RBO was comparable with that of hexane extraction (22.5%). The phytochemical contents of the RBO under the optimum conditions were in the range of tocols, 1500-1800 ppm; sterols, 15,350-19,120 ppm and oryzanol 5800-11,110 ppm.     

Determination of antioxidant components in rice bran oil extracted by microwave-assisted method

Rice bran oil was extracted by microwave-assisted extraction with isopropanol and hexane using a solvent-to-rice bran ratio of 3:1 (w/w). The experiments were done in triplicate at 40, 60, 80, 100, and 120 degrees C with a total extraction time of 15 min/sample. The oil components were separated by normal-phase HPLC and quantified with a fluorescence detector. The radical scavenging capability of the oil was tested with DPPH and was expressed as mumol Trolox Equivalent Antioxidant Activity. The increase in total vitamin E with temperature from 40 to 120 degrees C was 59.63% for isopropanol and 342.01% for hexane. Isopropanol was the best solvent for the extraction of gamma-tocopherol and gamma-tocotrienol as compared with hexane for both microwave-assisted and conventional solvent extraction. Isopropanol was better for oil yield extraction at high temperatures. Samples extracted with isopropanol at 120 degrees C had higher antioxidant activity. No differences in oil yield, total vitamin E, and antioxidant activity of oil was noticed between the two methods (microwave-assisted and solvent extractions), at 40 degrees C. No degradation of alpha-tocopherol was noticed during the process.     

Accelerated solvent extraction of monacolin K from red yeast rice and purification by high-speed counter-current chromatography

Monacolin K from red yeast rice was extracted by accelerated solvent extraction (ASE). The effects of various extraction parameters including extraction temperature, static extraction time and cycle index on yield were investigated using a DIONEX ASE 300 system to select the optimal conditions by an orthogonal test design L₉ (3)³. The optimum extraction conditions were determined as follows: extraction temperature 120 °C, static extraction time 7 min, and cycle index 3. Under the optimal conditions, the yield of ASE extract and monacolin K was 5.35% and 9.26 mg/g of dry red yeast rice, respectively. A separation and purification method of monacolin K was then established using high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane–ethyl acetate–methanol–water (8:2:5:5, v/v/v/v). From 300 mg of crude extract, 51.2 mg of monacolin K was obtained with the purity of 98.7%. The chemical structure of isolated compound was identified by UV, ESI-MS and ¹H NMR.     

Optimization of cyclotide extraction parameters

Cyclotides are gene-encoded plant mini-proteins that contain a unique circular and cystine knotted amide backbone. Because of that ultra stable scaffold and the ability to harness a wide variety of sequences and biological activities within the scaffold, cyclotides find interesting potential applications for drug discovery and in agriculture. However, some fundamental knowledge is still missing to exploit these plant compounds, including finding the optimal process of their extraction from plant material. In the current work, the extraction parameters solvent type, time of extraction, number of re-macerations and the plant material to solvent ratio have been compared using the sweet violet (Viola odorata L.) as a model plant. That species is a well-characterized and rich source of cyclotides that contains prototypic cyclotides with different chemical and physical properties. We found that hydroalcoholic solutions of medium polarity give good yield of the cyclotide cocktail. In conclusion, single maceration with 50% MeOH for 6 h at a plant material to solvent ratio of 0.5:10 (g/mL) represents an optimum extraction method.     

The Effect of Ultrasonificated Extracts of <Emphasis Type="Italic">Spirulina maxima</Emphasis> on the Anticancer Activity

The effect of ultrasonic extraction on extraction yields, cytotoxicity, and anticancer activity of Spirulina maxima was investigated in this study. Optimal extraction conditions were determined as 60 kHz frequency at 60°C for 30 min with 120 W intensity, which resulted in 19.3% of extraction yields and 19.1% of cytotoxicity on normal human cells. Yields from conventional water and ethanol extraction were 15.8% at 100°C and 8.3% at 80°C, respectively. It was found that the extracts obtained by ultrasonic extraction process selectively inhibited the digestive-related cancer cell lines, such as human stomach cancer cells, having 89% of the highest inhibition ratio and 4.5 of the highest selectivity. In adding 0.5 mg/mL of the extract, human promyelocytic leukemia cells' cell differentiation was increased 1.72 times over that of the control. Expression level of B cell lymphoma-2 from Hep3B cell was also effectively suppressed by the extract obtained at 60 kHz and 60°C, leading to the inhibition of the early step of carcinogenesis. This work suggests that anticancer activity of the extracts is due to water-soluble polysaccharides rather than proteins and is further supported by the result that the ultrasonification extraction process can efficiently extract relatively intact polysaccharides rather than digesting the proteins in S. maxima by matrix assisted laser desorption ionization-time of flight and high performance size exclusion chromatography chromatogram analyses. Therefore, ultrasonic extraction increases both extraction yield and the biological activity of S. maxima extracts, which might be useful as an alternative natural anticancer agent in the medical and food industries.     

一种测定女贞子中齐墩果酸和熊果酸的新方法

为建立一种加速溶剂萃取-毛细管区带电泳测定女贞子中齐墩果酸和熊果酸的新方法.考察了萃取温度、萃取时间和萃取次数对目标物萃取效率的影响,并考察了硼砂浓度,β-环糊精浓度、pH及甲醇浓度对目标物分离的影响.结果表明:(1)萃取温度和萃取次数影响目标物的萃取效率,而萃取时间的影响很小;萃取压力影响萃取过程的重现性.(2)优化的萃取条件为:萃取压力6.9 MPa,萃取温度100℃,萃取时间5 min和萃取次数2次.(3)优化的缓冲体系为:40 mmol/L硼砂,1 mmol/Lβ-环糊精,pH 9.5及6%甲醇.(4)齐墩果酸和熊果酸分别在10~200、10~160 mg/L范围内线性良好,相关系数均大于0.996,检测限分别为3.2 mg/L和3.0 mg/L,加标回收率为93%~97%.对比了加速溶剂萃取、索氏提取以及超声提取的提取效率.(5)不同提取方法比较结果表明,加速溶剂萃取的提取效率与索氏提取法接近,但高于超声提取;加速溶剂萃取法的主要优点是消耗提取溶剂量少,提取时间短,样品用量小.     

Effect of processing on the recovery of recombinant beta-glucuronidase (rGUS) from transgenic canola

This study addresses the processing of transgenic canola seed for production of recombinant proteins by using beta-glucuronidase (rGUS) as a model protein. The major processing steps that were investigated included dry and wet grinding of the seed, solvent extraction of canola oil, and protein extraction. rGUS in canola seed was stable for at least 2 weeks of incubation at 38 degrees C and for more than 5 months at 10 degrees C. At 70 degrees C, the residual activity changed inversely to the initial moisture content of the seed. The comparison of wet and dry processing revealed no significant differences in protein recovery. rGUS was stable during the defatting of transgenic canola flakes with hexane at 66 degrees C, whereas 2-propanol extraction at the same temperature reduced the extractable enzyme activity by almost 50%. The particle size of the ground seed was important for the extraction efficiency. A faster extraction and greater protein yield was achieved by extracting particles with an average diameter equal to or smaller than 255 microm. More than 80% rGUS was extracted in one stage with sodium phosphate buffer of pH 7.5.     

Bestimmung von Fumonisinen in komplexen Lebensmittelmatrices mittels Accelerated Solvent Extraktion (ASE)

Accelerated solvent extraction (ASE) is an alternative sample extraction procedure for fumonisins in corn and corn products. ASE gave results comparable to that of a draft CEN method, but required less extraction time. Furthermore, ASE gave significantly higher quantitative values than another method reported for extraction of fumonisins (Trucksess et al., 1995).     

Comparison of the capabilities of accelerated solvent extraction and sonication as extraction techniques for the quantification of kavalactones in Piper methysticum (Kava) roots by high performance liquid chromatography with ultra violet detection

A conventional extraction technique of sonication has been compared, in terms of extraction efficiency, extraction time and amount of solvent, with the more novel technique of accelerated solvent extraction for the extraction of kavain from the powdered roots of Piper methysticum (Kava) with acetone. The extracts were analysed using high-performance liquid chromatography with ultra violet detection. The effects of varying solvent volume and extraction time upon the quantity of kavain extracted with sonication, and the effects of varying temperature upon the kavain extraction efficiency by ASE, were investigated. ASE was found to be more efficient with respect to time and solvent volume required; however, a good agreement was found between the kavain concentration obtained using both extraction techniques.     

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.     

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.     

Extraktion von Ochratoxin A aus geröstetem Kaffee Mittels Accelerated Solvent Extraction (ASE)

Accelerated solvent extraction (ASE) is an alternative sample extraction procedure for ochratoxin A in roasted coffee. ASE results are comparable to that of the modified Koch method, but required less sample preparation time. Furthermore, ASE gave higher quantitative values than other methods reported for extraction of ochratoxin A. In the end less harmful water could be used for extraction.     

Optimisation of conditions for the extraction of casearins from Casearia sylvestris using response surface methodology

Optimal conditions for the extraction of casearins from Casearia sylvestris were determined using response surface methodology. The maceration and sonication extraction techniques were performed using a 3 x 3 x 3 full factorial design including three acidity conditions, three solvents of different polarities and three extraction times. The yields and selectivities of the extraction of casearins were significantly influenced by acidity conditions. Taking into account all variables tested, the optimal conditions for maceration extraction were estimated to involve treatment with dichloromethane saturated with ammonium hydroxide for 26 h. Similar yields and selectivities for casearins were determined for sonication extraction using the same solvent but for the much shorter time of 1 h. The best results for stabilisation of the fresh plant material were obtained using leaves that had been oven dried at 40 degrees C for 48 h.     

超声辅助低共熔溶剂提取沙棘籽粕多酚的工艺优化

以一系列低共熔溶剂为提取剂,采用超声波辅助法从沙棘籽粕中提取多酚。在单因素试验结果基础上,利用Box-Behnken实验设计,运用响应面分析法对影响沙棘籽粕多酚得率的主要因素（超声功率、超声时间、超声温度）进行优化。结果表明,沙棘籽粕多酚最佳提取工艺条件为：以含水量为30%的氯化胆碱-草酸低共熔溶剂为最佳提取剂,液料比为14∶1,超声功率420 W,超声时间56 min,超声温度44℃。在此条件下,多酚得率为3.31±0.008%。对比试验发现：氯化胆碱-草酸低共熔溶剂对沙棘籽粕多酚的得率明显优于传统溶剂;与热回流提取相比,超声提取法具有明显的优势。     

核桃楸叶片中总黄酮的最佳提取工艺研究

利用超声波法对核桃楸叶片中总黄酮的提取工艺进行了研究,在单因素试验的基础上,采用正交试验法,确立最佳提取条件,考察乙醇浓度、提取温度、液料比和提取时间对核桃楸叶片总黄酮提取率的影响。结果表明：超声波法辅助提取核桃楸叶片总黄酮的最适工艺参数是浸提剂乙醇浓度为70%,提取温度为50℃,液料比为20：1,提取时间为20 min,且总黄酮提取率可达4.458%。     

Extraction of chlorophylls and carotenoids from dry and wet biomass of isolated Chlorella Thermophila: Optimization of process parameters and modelling by artificial neural network

Chlorophylls and carotenoids can be extracted from microalgae using various solvents. However, there is lack of studies regarding the comparison of extraction yield of these pigments from wet and dry microalgal biomass using different combination of cell disruption methods. Therefore, in this work, we have investigated the comparison of the extraction yield of chlorophylls and carotenoids from the wet and heat-dried microalgal biomass (isolated Chlorella thermophila) using ethanol. Extraction parameters such as homogenisation time, homogenisation speed, solvent temperature, solid-solvent ratio, boiling time and microwave time have been optimised. Chlorophyll extraction yield was observed to be 2.7 fold higher from wet biomass than dry biomass while carotenoid yield was 6.7 fold higher. Highest chlorophyll yield (∼60 mg/g-dry biomass) was observed at 6 min of homogenisation time, 10,000 rpm, solid solvent ratio of 1 mg/mL and 58 °C of solvent temperature from wet biomass with extraction efficiency of ∼94 %. Highest carotenoid yield was noticed following the same conditions of chlorophyll extraction except 4 °C of solvent temperature. The modelling of the extraction process was performed using artificial neural network (ANN) which may be useful for the scale-up of the extraction process at the industrial level.     

Patterns of furanocoumarin production and insect herbivory in a population of wild parsnip (Pastinaca sativa L.)

Summary Seasonal changes in the distribution and abundance of furanocoumarins in wild parsnip, Pastinaca sativa (Umbelliferae), were examined in a population of plants in Tompkins County, New York. Xanthotoxin, imperatorin and bergapten (linear furanocoumarins) occur in all above-ground parts of the plant; in addition, angelicin and sphondin (angular furanocoumarins) occur in umbels of some individuals. Total furanocoumarin content, as measured by percent dry weight, is greatest in reproductive parts, particularly buds and seeds; variation in concentrations between plants is greatest in vegetative structures (e.g., leaves).Within the plant, the distribution of furanocoumarins is significantly correlated with nitrogen, as opposed to biomass, allocation. In that nitrogen is often a factor limiting the plant growth, furanocoumarins appear to be allocated in proportion to plant tissue value; reproductive structures, obvious contributors to plant fitness, contain over ten times the amount of nitrogen and furanocoumarin contained in vegetative structures such as senescent leaves.Stepwise multiple regression analysis revealed that generalized insect herbivores tend to feed on plants or plant parts low in furanocoumarin content and, correspondingly, low in nitrogen content. Parsnip specialists, notably Depressaria pastinacella (Lepidoptera: Oecophoridae), feed exclusively on umbels, plant parts rich in nitrogen and furanocoumarins; furanocoumarin number and content in fact account for over 60% of the variance in number of umbel feeders. These patterns conform with previous determinations of the toxicological properties of furanocoumarins. Nitrogen is known to affect growth rate, fecundity, longevity and survivorship of insect herbivores; by tolerating or detoxifying furanocoumarins, D. pastinacella can consume plant tissues containing significantly greater amounts of nitrogen than tissues consumed by generalist feeders. That the presence of D. pastinacella on individual plants is correlated with the number of furanocoumarins present is consistent with the hypothesis that parsnip specialists use angular furanocoumarins as host recognition cues.     

Development of a liquid chromatography-tandem mass spectrometry with pressurized liquid extraction method for the determination of benzimidazole residues in edible tissues

A confirmatory and quantitative method of liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with a pressure liquid extraction (PLE) was developed for the determination of 11 benzimidazole and 10 metabolites of albendazole, fenbendazole and mebendazole in the muscles and livers of swine, cattle, sheep and chicken. For sample preparation, we used an automated technique of PLE method. The optimum extraction conditions were obtained using an 11 ml Accelerated Solvent Extraction (ASE) cells, acetonitrile/hexane as the extraction solvent. HPLC analysis was performed on a C18 column with gradient elution using acetonitrile and 5 mmol l(-1) formic ammonium as mobile phase. The analytes were detected in the positive ion multiple reaction monitoring (MRM) mode by the LC-ESI-MS/MS analysis. The recoveries of benzimidazole (BZDs) spiked at the levels of 0.5 μg kg(-1) ranged from 70.1% to 92.7%; the between-day relative standard deviations were no more than 10%. The limits of quantification were 0.02-0.5 μg kg(-1). The optimized method was successfully applied to monitor real samples containing BZDs, demonstrating the method to be simple, fast, robust and suitable for identification and quantification of BZDs residues in animal products.