微藻食用油脂浸提溶剂的选择与分析

分别向微藻中加入6种常用溶剂,冰浴超声处理15 min,振荡浸提8 h,提取微藻中的油脂,考察了溶剂的浸提能力.结果表明,浸提干藻粉与湿藻泥中的油脂可选择不同溶剂,用正己烷浸提干藻粉中油脂,提取率可达71.52%,其中中性脂占75.86%,维生素含量E为1.63%,提取油脂后的藻渣中蛋白质、叶绿素和糖类的保留率分别为55.92%,61.33%和78.35%,营养成分保留率较高;用正己烷/乙醇混合溶剂浸提湿藻泥中油脂,提取率可达68.31%,其中中性脂占71.65%,维生素E含量为1.87%,提取油脂后的藻渣中蛋白质、叶绿素和糖类的保留率分别为60.56%,53.27%和80.20%.     

微拟球藻油脂萃取及脱脂藻水热液化

为提高微藻的综合利用效率,使用不同的溶剂系统分别对干、湿微拟球藻进行油脂萃取,并对脱脂后的藻渣进行水热液化实验,探究溶剂萃取脱脂对微藻水热液化产物的影响。溶剂萃取的结果表明,极性溶剂对油脂的萃取率达到25.0%,但对脂质的萃取缺乏选择,萃取物的脂肪酸甲酯产率仅为29.68%;混合溶剂萃取的脂肪酸甲酯回收率达到57.70%。脱脂后的微拟球藻水热粗油产率为27.7%~34.6%,氮含量为5.29%~6.68%,主要由脂肪酸、脂肪酸酯、脂肪酸酰胺、长链烃类、胺类、含氧化合物和含氮杂环化合物组成。经甲醇萃取后的湿藻水热粗油产率为34.6%,氮含量为5.44%,过程能耗低,表明甲醇萃取湿藻结合水热液化具有一定的应用前景。     

微拟球藻油脂萃取及脱脂藻水热液化

为提高微藻的综合利用效率,使用不同的溶剂系统分别对干、湿微拟球藻进行油脂萃取,并对脱脂后的藻渣进行水热液化实验,探究溶剂萃取脱脂对微藻水热液化产物的影响。溶剂萃取的结果表明,极性溶剂对油脂的萃取率达到25.0%,但对脂质的萃取缺乏选择,萃取物的脂肪酸甲酯产率仅为29.68%;混合溶剂萃取的脂肪酸甲酯回收率达到57.70%。脱脂后的微拟球藻水热粗油产率为27.7%～34.6%,氮含量为5.29%～6.68%,主要由脂肪酸、脂肪酸酯、脂肪酸酰胺、长链烃类、胺类、含氧化合物和含氮杂环化合物组成。经甲醇萃取后的湿藻水热粗油产率为34.6%,氮含量为5.44%,过程能耗低,表明甲醇萃取湿藻结合水热液化具有一定的应用前景。     

从雨生红球藻藻泥中选择性提取虾青素

以雨生红球藻湿藻泥为原料,研究了不同有机溶剂对胞内油脂和虾青素选择性提取分离的影响,通过酸解破壁提高虾青素和油脂的提取效率。结果表明,连续乙醇提取可对胞内色素和油脂有效分级提取,先提取出极性组分(叶绿素和极性脂),再提取中性组分(类胡萝卜素和中性脂)。中等极性溶剂或溶剂体系对类胡萝卜素的选择性和提取率较好;乙醇/乙酸乙酯混合溶剂提取类胡萝卜素的总得率(干重)达25.31 mg/g,提取率为69.35%。对雨生红球藻湿细胞进行酸解破壁处理有助于提高虾青素和油脂的提取率。在最优酸解破壁条件(盐酸浓度1 mol/L,温度60℃,时间60 min)下,含水80%的雨生红球藻藻泥的油脂总得率(干重)达418 mg/g,总脂提取率达97%。     

溶剂提取-荧光光谱法测定微藻油脂

研究了溶剂提取-荧光测定微藻油脂的一种新的方法。通过与溶剂提取-水浴蒸干称重法比较,表明溶剂提取-荧光光谱法具有快速、简便、高效、灵敏、不受藻样不能长期保存而影响测定制约的特点,可测定藻体量较低时的低浓度油脂含量,而称重法不能测得。利用藻密度、藻干重、藻液吸光度、荧光强度、油脂之间良好的线性关系,通过溶剂提取-荧光光谱法,测得打捞的微藻油脂平均含量为10.45%,溶剂提取-水浴蒸干称重法测得微藻油脂平均含量为8.17%,结果表明溶剂提取-荧光测定法的结果比溶剂提取-水浴蒸干称重法测得值偏大,能更准确地反映真实数值。     

萃取湿藻油脂及CO_2切换溶剂的应用

藻类被认为是油脂提取的一种重要的可持续的原料来源,提取的油脂可用于生产食品、化妆品、药物和生物燃料。除去种植成本,这一过程尤其受萃取前干燥微藻和之后溶剂回收的能量消耗所限制。本文综述了传统的从微藻(特别是湿藻)中萃取油脂的技术(有机溶剂萃取和超临界二氧化碳萃取)以及CO2切换溶剂的研究进展,并指出了CO2切换溶剂在藻类油脂萃取中的应用前景。     

利用气相色谱-质谱联用技术测定微藻油脂的组成

采用超临界CO2萃取技术从微藻粉中提取油脂,微藻油脂经预处理后,采用气相色谱-质谱联用技术分析微藻油脂的组成成分.测定结果显示微藻油脂中主要有20种组分,主要是C12到C24的化合物,其中酯类化合物较多,以邻苯二甲酸二异辛酯居多,质量分数占70％以上,其次分别为反式角鲨烯、软脂酸等.     

利用有机溶剂提取微藻油脂的方法探究

在传统化石能源日益枯竭的趋势下,微藻生物柴油作为第三代绿色可再生的替代型能源越来越受到人们的重视.在微藻生物柴油的产业链上,油脂的提取是影响其推广应用的一个关键环节.本文实验利用有机溶剂提取微藻油脂,探究在不同的条件下微藻油脂的提取效果,并特别研究了先后使用甲醇和石油醚两种有机溶剂对微藻油脂提取率的影响.研究结果表明:温度、液料比、浸提时间对提取效率都有一定的影响,并且使用甲醇和石油醚两种溶剂分步提取时会使微藻油脂提取率明显提高;在液料比为15mL/g、提取温度为45℃、提取时间为5h时,使用石油醚作为提取剂的提取率为58.71%;使用甲醇溶剂提取后再使用石油醚提取时,在液料比和提取时间相同的条件下,温度为35℃时提取率即可达87.90%     

利用微藻净化生活污水的可行性研究

探讨利用微拟球藻对污水和中水进行深度净化的可行性,从而获得污水净化的最佳条件。结果表明,该藻对污水氮磷具有较强的净化能力,可在13天内去除水体中96%的氨氮和94%的磷,同时COD的去除率可以达到72.9%。在生活污水中培养至第16天,微藻的细胞密度每毫升可达4.55×10~6个。该研究以中水和生活污水为基质培养微拟球藻,为污水中氮磷的去除和能源的同步获取提供了新途径。     

亚临界流体萃取法回收废弃虾蟹壳中虾青素

考察了以废弃虾蟹壳为原料,亚临界流体萃取法提取虾青素的工艺。通过单因素及正交实验优化了提取条件,提取工艺为:以二氯甲烷为溶剂,100℃,9.31~11.72 MPa,静态提取时间15 min,循环2次,提取率为0.037 3%。与传统的有机溶剂浸提法提取结果比较表明,亚临界流体萃取的提取率高出33.2%,且耗时减少82.8%。     

Subcritical co‐solvents extraction of lipid from wet microalgae pastes of Nannochloropsis sp.

In this paper subcritical co‐solvents extraction (SCE) of algal lipid from wet pastes of Nannochloropsis sp. is examined. The influences of five operating parameters including the ratio between ethanol to hexane, the ratio of mixed solvents to algal biomass (dry weight), extraction temperature, pressure, and time were investigated. The determined optimum extraction conditions were 3:1 (hexane to ethanol ratio), 10:1 ratio (co‐solvents to microalgae (dry weight) ratio), 90°C, 1.4 MPa, and 50 min, which could produce 88% recovery rate of the total lipids. In addition, electron micrographs of transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were conducted to show that the algal cell presented shrunken, collapsed with some wrinkles and microholes after SCE extraction. The main composition of total lipids extracted under the optimum conditions was TAG which represented more than 80%. And the fatty acid profile of triglycerides revealed that C16:0 (35.67 ± 0.2%), C18:1 (26.84 ± 0.044%) and C16:1 (25.96 ± 0.011%) were dominant. Practical applications: The reported method could save energy consumption significantly through avoiding deep dewatering (for example drying). The composition of the extracted lipid is suitable for the production of high quality biodiesel.     

Optimization of algal lipid extraction by mixture of ethyl acetate and ethanol via response surface methodology for biodiesel production

The effects of extraction time, extraction temperature, solvent to biomass ratio and solvent composition on lipid yield from lyophilized Chlorococcum sp. biomass using a mixture of ethyl acetate and ethanol (EAE), a new proposed solvent, were studied. Subsequently, the process conditions of extraction by EAE were optimized using Box-Behnken design (BBD). The results revealed that the extraction temperature had the greatest effect on lipid extraction efficiency, followed by volume ratio of ethyl acetate to ethanol (EA/E) and extraction time. The largest lipid extraction yield of 15.74% was obtained under the following extraction conditions: 40mL solvents per gram of biomass for 270 min with gentle stirring at 80 °C by EAE with an EA/E of 1.0. Furthermore, palmitic acid, stearic acid, oleic acid, and linoleic acid were the most abundant fatty acids in the lipids extracted, indicating the great potential of the proposed lipid extraction procedure for microalgae-based biodiesel production.     

Extraction of lipids from fermentation biomass using near-critical dimethylether

The extraction of lipids from both wet and dry biomass produced by fermentation has been carried out using near-critical dimethylether (DME) as the extraction solvent. Fermentations were carried out from a shake flask up to a 300 L scale using the microorganism Mortierella alpina, and up to a 20 L scale for Phaffia rhodozyma and Agrobacterium tumefaciens. The lipids extracted at a laboratory and pilot scale from the biomasses were enriched in arachidonic acid, astaxanthin, and co-enzyme Q10 respectively. Extractions were also performed on marine microalgae, produced by a proprietary fermentation process, to obtain lipids rich in EPA. Lipids were extracted from wet biomass using DME, which removes the need to dry the biomass. Water is also co-extracted, which has to be separated from the lipid. The biomass shrunk considerably during packed bed extraction of wet biomass, leading to channelling. Repacking and re-extraction of the packed bed enabled full lipid yields to be obtained. The extraction of lipids from biomass suspended in fermentation broth showed considerable promise, and lipid yields were improved due to the recovery of lipids that had been exuded into the broth from the microorganism. In contrast, the extraction of lipids from freeze-dried biomass using DME was routine, yields were substantially higher than using CO₂ or CO₂ + ethanol, but were lower than from wet biomass. DME also extracted polar lipids from both wet and dry biomass, leading to the higher total lipid yields compared to CO₂. Separate extraction of non-polar and polar lipids was possible by sequential extraction of dry biomass using initially CO₂ followed optionally with ethanol co-solvent; and then DME.     

Optimization of fatty acid extraction from Phaeodactylum tricornutum UTEX 640 biomass

Fatty acids in the microalga Phaeodactylum tricornutum were isolated using an optimized three-step method: extraction of crude fatty acid potassium salts made by direct saponification of lipids in the microalgal biomass with KOH/ethanol (96%, vol/vol), separation of unsaponifiable lipids by extraction with hexane, and final purification of fatty acids by acidification of the alcoholic solution of potassium soaps followed by extraction of fatty acid into hexane. Direct saponification was carried out in ethanol (96%, vol/vol) using 2.09 mL ethanol (96%) per gram of wet biomass (10 mL/g of dry biomass) mixed with 0.4 g KOH/g of biomass. Under these conditions the fatty acid yield was 87%. The optimal water content of the alcoholic solution for extraction of the unsapononifiables was established as 40%, w/w. Data on equilibrium carotenoid distribution between the alcoholic (40%, w/w water) and hexane phases were determined. These data allow prediction of the carotenoid yields with different volumes of hexane in several extraction steps. The optimal pH of the alcoholic solution before extracting the purified fatty acid was established as pH 6, and the equilibrium fatty acid distribution between the alcoholic and hexane phases was determined. This optimized method permited a 20% reduction in the production costs of highly purified eicosapentaenoic acid (EPA) in the three-step preparative process (extraction of fatty acid, concentration of polyunsaturated fatty acids by the urea method, and EPA fractionation through preparative high-performance liquid chromatography) previously developed by the authors.     

Influence of pretreatments for extraction of lipids from yeast by using supercritical carbon dioxide and ethanol as cosolvent

Saccharomyces cerevisiae is one of the most studied and industrially exploited yeast. It is a non-oleaginous yeast whose lipids are mainly phospholipids. In this work, the extraction of yeast lipids by supercritical carbon dioxide (SCCO₂) and ethanol as a co-solvent was studied. In particular our attention was focused on the selectivity toward triglycerides, and in a subsequent extraction of the phospholipids present in the yeast. Indeed CO₂ is a non-polar solvent and is not an efficient solvent for the extraction of phospholipids. However, SCCO₂ can be used to extract neutral lipids, as triglycerides, and the addition of polar co-solvents like ethanol, at different compositions, allows a more efficient extraction of triglycerides, and also an extraction-fractionation of phospholipids. In this work SCCO₂ extractions of a specific membrane complex of S. cerevisiae, obtained from an industrial provider, were carried out at 20 MPa and 40 °C, using ethanol as a co-solvent (9%, w/w). It was shown that different pretreatments are necessary to obtain good extraction yields and have a great impact on the extraction. The kinetic of the extractions were successfully modeled using Sovova's model. From the fitting of the main parameters of the model it was possible to compare the effects of the pretreatments over the yeast material, and to better understand the extraction process. Among the seven tested pretreatments the more appropriate was found to be an acid hydrolysis followed by a methanol maceration.     

Ultrasonic extraction of ferulic acid from Ligusticum chuanxiong

The extraction of ferulic acid, a pharmacologically active ingredient from the root of Ligusticum chuanxiong, with ultrasonic extraction was investigated. Percolation and supercritical fluid extraction (SFE) were employed to make comparisons with ultrasonic extraction. Three variables, which included the concentration of solvent, the ratio of solvent volume/sample (mL/g), and extraction time, were found to have a great influence on ultrasonic extraction. The optimum extraction was with pure ethanol with a solvent volume/sample ratio 8:1 (mL/g) and extraction time of 30 min. Under the optimum extraction conditions, the extraction yield could reach 8.8% which was higher than that using SFE with ethanol as co-solvent and a content of ferulic acid of 0.7%; both the yield and the content were higher than those obtained by percolation. Ultrasonic extraction significantly shortened the time required for the extraction process. Overall, ultrasonic extraction was shown to be highly efficient in the extraction of ferulic acid from Ligusticum chuanxiong.     

Optimization of cell wall disruption and lipid extraction methods by combining different solvents from wet Chlorella vulgaris

Over the past few decades, microalgae have emerged as a promising option for making lipid-based bioactive compounds. The purpose of this study was to extract lipids from Chlorella vulgaris. In this study, different cell wall disruption methods such as microwave, liquid nitrogen, ultrasound (US), and bead mill were compared. We selected and optimized two systems based on their efficacy in disrupting cell walls—US and beads mill. Based on the dry weight of C. vulgaris biomass, the maximum lipid extraction by the US was 17.1% and by bead mill was 15.2%. Following cell wall disruption of C. vulgaris, chloroform–methanol (2:1) solvent combination achieved high lipid extraction. However, the hexane–ethanol (1:1) solvent combination was chosen because of its lower toxicity. Specifically, the effect of the solvent-to-biomass ratio, the temperature, and the extraction time was investigated. The results indicated that the chloroform–methanol solvent combination yielded optimal results at 8 ml/g solvents to biomass, 45°C, and 60 min and that the hexane–ethanol combination yielded optimal results at 6 ml/g, 35°C, and 73 min, respectively. The highest amount of lipids was obtained from C. vulgaris with 87.6% moisture content. As a cell wall disruption method, the US obtained 20.4% and 16.4% with a combination of chloroform–methanol solvents and hexane–ethanol, respectively. Additionally, bead milling resulted in the highest extraction yield of 17.6% for chloroform–methanol and 13.9% for hexane–ethanol. Based on the results of cell wall disruption, the US method is the most efficient cell wall disruption method in terms of lipid extraction efficiency.     

Evaluating Cell Disruption Strategies for Aqueous Lipid Extraction from Oleaginous Scenedesmus obliquus at High Solid Loadings

Among various renewable energy sources, the production of biofuels derived from algal lipids holds bright prospects. One of the major roadblocks in the successful commercialization of microalgal biofuels is the existing energy‐intensive lipid extraction. In the present investigation, an attempt is made to assess aqueous lipid extraction strategies from oleaginous Scenedesmus obliquus at a high solid loading of 15% (w/v). In this study, four surfactants and five enzymes are evaluated for cell disruption of S. obliquus. It is the first report citing cetyl pyridinium bromide as the most suitable cationic detergent for surfactant‐assisted extraction, with a lipid recovery as high as 31.4%. However, during the evaluation of enzyme‐based cell disruption, neutral protease emerges as the best biocatalyst resulting in a lipid recovery of ≈75%. Total lipid extraction is accomplished using a two solvent system comprising of water‐immiscible ethyl acetate, followed by chloroform addition. The study revalidates the fact that the biochemical composition of Scenedesmus sp. plays a vital role while identifying and formulating an efficient and green process for microalgal cell disruption for enhanced lipid extraction under aqueous conditions. Practical Applications: The results of the present study demonstrate that if the biochemical composition of any oleaginous algal cell wall is known, aqueous enzymatic lipid extraction can be employed rather than taking up the conventional route of drying followed by Soxhlet extraction. The combination of using the cheap sources of enzymes and water‐immiscible green solvents like ethyl acetate can be lucrative downstream procedures for the lipid recovery from wet algal biomass when compared to traditional procedures.     

Extraction of Algal Lipids and Their Analysis by HPLC and Mass Spectrometry

Algae are a promising source of biofuel but claims about their lipid content can be ambiguous because extraction methods vary and lipid quantitation often does not distinguish between particular lipid classes. Here we compared methods for the extraction of algal lipids and showed that 2-ethoxyethanol (2-EE) provides superior lipid recovery (>150–200 %) compared to other common extraction solvents such as chloroform:methanol or hexane. Extractions of wet and dry algal biomass showed that 2-EE was more effective at extracting lipids from wet rather than dried algal pellets. To analyze lipid content we used normal-phase HPLC with parallel quantitation by an evaporative light scattering detector and a mass spectrometer. Analysis of crude lipid extracts showed that all major lipid classes could be identified and quantified and revealed a surprisingly large amount of saturated hydrocarbons (HC). This HC fraction was isolated from extracts of bioreactor-grown algae and further analyzed by HPLC/MS, NMR, and GC/MS. The results showed that the sample consisted of a mixture of saturated, straight- and branched-chain HC of different chain lengths. These algal HC could represent an alternative biofuel to triacylglycerols (TAG) that could feed directly into the current petroleum infrastructure.