WO2000005395A1 - Verfahren zur präparativen gewinnung von fettsäuren aus biomasse durch in- situ-extraktion-reaktion-chromatographie mit verdichteten gasen - Google Patents
Verfahren zur präparativen gewinnung von fettsäuren aus biomasse durch in- situ-extraktion-reaktion-chromatographie mit verdichteten gasen Download PDFInfo
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- WO2000005395A1 WO2000005395A1 PCT/EP1999/005107 EP9905107W WO0005395A1 WO 2000005395 A1 WO2000005395 A1 WO 2000005395A1 EP 9905107 W EP9905107 W EP 9905107W WO 0005395 A1 WO0005395 A1 WO 0005395A1
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- reaction
- extraction
- fatty acid
- fatty acids
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Classifications
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C3/00—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom
- C11C3/003—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom by esterification of fatty acids with alcohols
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
- A23L33/12—Fatty acids or derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
- A61K31/23—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of acids having a carboxyl group bound to a chain of seven or more carbon atoms
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
- C11B1/104—Production of fats or fatty oils from raw materials by extracting using super critical gases or vapours
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B7/00—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
- C11B7/0008—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents
- C11B7/005—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents in solvents used at superatmospheric pressures
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C3/00—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom
- C11C3/04—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom by esterification of fats or fatty oils
- C11C3/10—Ester interchange
Definitions
- the invention relates to a method for the preparative production of fatty acid esters - for the production of fatty acids, preferably polyunsaturated fatty acids (PUFA) - from biological sources by means of a continuous in situ extraction-reaction chromatography with compressed gases.
- fatty acids preferably polyunsaturated fatty acids (PUFA) - from biological sources by means of a continuous in situ extraction-reaction chromatography with compressed gases.
- PUFA polyunsaturated fatty acids
- PUFAs occur in nature in relatively high concentrations in linseed oil, nut oil, poppy seed oil, hemp oil and in fish oils. Numerous attempts have already been made to obtain these valuable substances from such biological sources and to isolate them in more or less high purity. However, since the PUFAs are usually chemically bound as esters in lip (o) iden, in addition to extraction, they must be converted into the free acids (hydrolysis) or into corresponding monoesters (transesterification).
- Lip (o) ide mostly contain fatty acids bound in glycerides (mono-, di-, tri-glycerides), phosphatides, glycolipids and aminolipids. These bound fatty acids as well as free native fatty acids and their derivatives differ on the one hand in the frequency of their occurrence in biological sources, and on the other hand in their effect on the human organism.
- native fatty acids and their derivatives are obtained from biological sources. especially by extraction with the help of compressed gases (eg supercritical carbon dioxide, etc.).
- compressed gases eg supercritical carbon dioxide, etc.
- SFE supercritical fluid extraction
- Organic acids serve as catalysts here, "Coulping chemical derivatization reactions with supercritical fluid extraction", JA Fields, J. Chromatogr. A, 785 (1997), pp. 239-249) and solid catalysts (e.g. ion exchange resins (C. Vieville, Z. Mouloungui, A. Gaset; Colloq. - Inst. Natl. Rech. Agron. (1995), 71 (Valorisations Non-Alimentraires des Grandes Productions Agricoles), 179-82 .; acidic aluminum oxide ( BW Wenclawiak, M. Krappe, A. Otterbach; J. Chromatogr. A (1997), 785, 263-267)) or combinations thereof (C. Vieville, Z. Mouloungui, A. Gaset; Ind. Eng. Chem. Res (1993), 32 (9), 2065-8).
- ion exchange resins C. Vieville, Z. Mouloungui, A. Gaset; Collo
- Enzymatic reactions by means of lipases are also known, which carry out the fat cleavage either in solution or immobilized with subsequent extraction in the presence of supercritical gases (R. Hashizume, Y. Tanaka, H. Ooguchi, Y.Noguchi, T. Funada; JP-196722 and A. Marty, D. Combes, JS Condoret; Prog. Biotechnol. (1992), 8 (Biocatalysis in Non-conventional Media), 425-32).
- the SFE and SFR processes can be carried out as a continuous (continuous flow) or discontinuous (batch) process.
- King et. al carry out extraction and transesterification in the presence of compressed gases as a batch process (J.W. King, J.E. France, J.M. Snyder; Fresenius J. Anal. Chem. (1992), 344, 474-478).
- lipids are first extracted from a biological source (here: seeds), followed by transesterification on the catalyst to methyl esters.
- the alumina catalyst is physisorbed with methanol.
- the complex insertion of the samples onto the catalyst is disadvantageous, as a result of which a reaction only takes place at the catalyst / sample interface. This is insufficient in the context of a preparative implementation.
- the present invention has for its object to provide a method for the preparative preparation of unsaturated and saturated fatty acid esters and their selective isolation from biological sources.
- the object is achieved in that a method for producing and isolating fatty acid esters from biological sources is provided by means of continuous in-situ extraction-reaction chromatography. In the presence of a compressed gas stream and a 0.5 to 5% C1-C5 alcohol modifier
- reaction is carried out on an inert catalyst in full contact with the biological source
- reaction products are chromatographed on the inert catalyst from (a), which has chromatographic retention and only desorbs and elutes the reaction products;
- reaction products produced are harmless to health and food chemistry, because
- Carbon dioxide can be used as the reaction / extraction medium or mobile phase. Neither the starting materials nor the products thus come into contact with toxic substances at any point in the process.
- Carbon dioxide serves as a protective gas atmosphere to prevent oxidation and gentle extraction and elution.
- the toxicity of the substances used in the present case is so low that they can be safely used for the production of food additives or pharmaceutical products.
- reaction products are obtained in pure form as a substance or in high concentrations in a suitable solvent and can easily be processed further.
- the reaction products are selectively separated from the starting materials, the intermediate or by-products in the process according to the invention.
- the process can be carried out continuously. In the case of liquid educts, they can be fed into the flow system.
- the biological source can be used directly. Limiting the amount of biomass from the biological source is not necessary and is therefore suitable for large-scale use.
- the process combines and combines the extraction, reaction and chromatography process stages to form a functional unit and can therefore reduce the costs for large-scale use.
- “In situ extraction-reaction chromatography” in the sense of this invention means that the fatty acid esters from the biological source are provided, chromatographed and extracted from the biological source by means of compressed gases (in short: SF-REC).
- the catalyst according to the invention serves as a stationary phase in which the fatty acid esters produced are selectively desorbed from the catalyst and eluted in the presence of the compressed gas. Therefore, the inventive Moderate catalyst have a chromatographic retention in which the product does not adsorb.
- the parameters (conditions) according to the invention are selected such that the lip (o) ide remain on the catalyst and the fatty acid esters are selectively eluted. These specific parameters are explained in the examples.
- the method of in situ reaction extraction chromatography according to the invention is carried out as a continuous method. It can also be used synonymously with the term “(continious) batch flow method”.
- the term continuous method is understood as a flow system in which, under continuous reaction in the gas / modifier stream, the reaction products on the solid or consistent biological
- the source in contact with the inert catalyst is chromatographed and extracted
- a liquid biological source can be added to the gas / ethanol stream and ensures a continuous process.
- compressed gases in the sense of this invention encompasses liquid, supercritical and two-phase or subcritical gases or gas mixtures.
- gases which are known to the person skilled in the field of SFE and SFR technology are expressly included here compressed gas for extraction and is used as a reaction medium
- the gas also serves as a mobile phase and is used as an extraction medium, and compressed carbon dioxide is particularly preferred.
- modifier means an additional stream in the presence of the compressed gas.
- 0.5-5% by volume of lower alcohols are used.
- An ethanol modifier of 0.5-5% by volume is preferred.
- the modifier serves as a reactant for the transesterification of lip (o) ides on the inert catalyst.
- fatty acid esters are obtainable from all branched and unbranched fatty acids as well as fatty acid derivatives, such as hydroxy fatty acids, which have a carbon chain of at least 12 carbon atoms.
- Fatty acid esters are preferred as fatty acid ethyl esters, since the alcohol required for the preparation has the lowest toxicity of the lower alcohols.
- the invention can be carried out for unbranched or branched C1-C5 alcohols.
- Educts are preferably lip (o) ide from biological sources and other bound fatty acids; Reaction products are fatty acid esters. Since ethanol is preferably used as the reactant and modifier for the reaction (transesterification), fatty acid ethyl esters are preferably obtained. Of course, free fatty acids and their salts are converted into their fatty acid esters in the biological sources.
- complete conversion means the conversion of all the fatty acids contained in the starting materials into the corresponding fatty acid ethyl esters while the unreacted starting materials remain (adsorption) on the inert catalyst.
- Inert catalyst in the sense of this invention means that in full contact with the biological source, this catalyst on the one hand accelerates the reaction (transesterification) into the reaction products and on the other hand serves as a stationary phase. For this, the catalyst must have chromatographic retention. The catalyst must not be toxic to the biological source and is therefore available to the reaction in an inert manner.
- Alumina which is commercially available, has proven to be advantageous and inexpensive, which can also be easily mixed and / or rubbed with the biological source, possibly with other auxiliaries and additives (e.g. sea sand).
- any biological source can be used and used in the method according to the invention.
- the term biological source is therefore preferably to be applied to easily cultivable microorganisms.
- Preferred are microorganisms with a high content of highly unsaturated fatty acids (PUFA) that can be easily broken down. nen. Be it chemical, enzymatic, but preferably mechanical.
- Microorganisms with a fatty acid spectrum which predominantly contain one or only a few bound or native fatty acids are particularly preferred. Organisms are preferred here, as follows:
- Bacillariophyta genus Nitzschia, Navicula, Cyclotella
- FIG. 1 describes the schematic structure and arrangement of the functional units, the terms having the following meaning:
- Catalyst steel chamber filled with alumina. Location of in situ extraction-reaction chromatography.
- Restrictor valve or narrow capillary that opposes a flowing medium and regulates the pressure system in the presence of the "CO 2", “sample”, “ethanol” pump.
- Alcohol 0.5 vol% to 5 vol% ethanol
- Catalyst separation medium 7g aluminum oxide A for column chromatography (ICN, acidic, activity I) (possibly also other, e.g. basic, neutral, coated with acids, different grain sizes or also on silica gel, zirconium oxide or other basis)
- ICN organic compound
- activity I possibly also other, e.g. basic, neutral, coated with acids, different grain sizes or also on silica gel, zirconium oxide or other basis
- the lower chromatogram in FIG. 2 shows the analysis (on aminopropyl phase 4.6x500mm, 10 ⁇ m; 5% ethanol as modifier, 150 bar 40 ° C.) of the substances originating from the system during the process. Under the given conditions (1% EtOH) only the ethyl ester is removed from the system. The upper chromatogram shows the substances removed from the system with the addition of 10% EtOH. Under these conditions, all substances (except glycerin) are eluted from the system. This is tantamount to stopping the reaction. You can see peaks for the ethyl ester, the triglyceride and the corresponding mono- and diglycerides
- Trap trap with glass balls (0.25 - 0.5 mm);
- GC-MS is used for qualitative analysis and for the identification of the products.
- SFC is used for quantification.
- a qualitative analysis of the fatty acid ethyl esters is particularly important for determining the fatty acid spectrum for the real samples. This is examined using GC-MS. For this purpose, the esters dissolved in n-heptane were separated on a DB5 column and identified by their mass spectra. The exact chromatographic conditions are listed in the appropriate place.
- the quantitative analysis of the ethyl esters formed and the unreacted triglycerides is used to determine the yield of the SF-REC.
- the problem arises of being able to determine all products (ethyl esters, mono- and diglycerides; not glycerol) and the starting material (triglyceride) using a chromatographic method.
- ethyl esters mono- and diglycerides; not glycerol
- triglyceride chromatographic method.
- only volatile esters methyl or ethyl esters
- the glycerides must therefore first be converted into the esters of lower alcohols.
- the problem of UV detection arises in HPLC.
- the glycerides and the ethyl esters must e.g. be converted into the phenacyl esters.
- SFC on amino phases and ethanol as modifiers, the ethyl esters and the glycerides can be separated and also detected underivatized at
- the SF-REC was carried out on the commercial HEWLETT-PACKARD system SFE Module 7860T.
- the conversion rate was checked after certain time intervals: after the start of the reaction, ie the introduction of the sample into the reaction vessel with catalyst and setting of the conditions (pressure, temperature, flow, etc.) on the device, were carried out continuously over a certain period of time the substances removed from the reaction vessel are collected.
- This collection process by means of a trap with solid sorbent is carried out in two steps: The extraction of the substances from the reaction chamber takes place in a so-called dynamic step, ie the extraction medium flows through the system, dissolves the sample and separates it after the fluid has been released , on the solid support of the trap.
- a second step this is rinsed with a suitable solvent (here: n-heptane) and the extracted substances are transferred to a collecting vessel.
- a suitable solvent here: n-heptane
- This rinsing step takes about six minutes. During this time the flow of the fluid is stopped, but the pressure and temperature in the The reaction chamber is retained. The reaction continues during this time. This step can therefore be called an additional so-called static step.
- the trap is rinsed after certain times (see above) to determine how much product has formed after this time. If the abscissa of the course of the reaction is labeled "time [min]" in the following, this means that only the dynamic steps have been added up. Two different reactions with the same sequence of steps are therefore comparable.
- the labeling of the time axis with "total reaction time [min]” means that the six minutes of the static course of the reaction during the rinsing process were added to each dynamic step. This should be taken into account when comparing the kinetics recorded!
- the collecting vessels are filled automatically.
- the amount of the n-heptane solution can be determined by weighing the vials before and after the step.
- the respective vials for each reaction step are injected three times for the quantification.
- the mean value of the peak areas of the three injections is formed for evaluation.
- the concentration is calculated from the calibration line and, with the amount of solution, the amount of substance.
- the theoretically achievable amount of oleic acid ethyl ester (EA) is calculated from this, which corresponds to 100% reaction yield.
- the determined amount of 18: 1 EE is based on this.
- the conditions for the transesterification using SF-REC can be varied within a wide range.
- the reaction or extraction conditions such as pressure, flow rate, modifier content and temperature can be independent of each other can be varied. This provides a number of parameters for optimizing the yield. If you consider the apparatus structure ( Figure 1), the individual points result as follows: Table 1 :
- the trap can be cooled to prevent the sample from evaporating or to condense, increase its viscosity, or freeze it. It should be noted that "blowing off" the sample in the form of droplets from the trap material must be prevented by the rapid gas flow. This point is particularly problematic when working with modifiers. This also separates out as a liquid. Since the amount of liquid exceeds the absorption capacity of the trap, there are losses because the sample dissolved in the modifier is "blown" by the trap. Therefore, when modifier is added, the trap must be heated above the boiling point of the modifier, so that it is gaseous like C0 2 and can no longer dissolve the sample. At this temperature, however, the volatility of the sample can lead to losses due to evaporation.
- the sample After being caught on the trap, the sample is eluted from the solid sorbent in a rinsing step (rinse) with a suitable solvent (rinse solvent).
- the second option is to introduce the relaxed gas, in which small sample droplets or particles are entrained, into a suitable solvent.
- the sample can dissolve in the solvent if it has enough time to come into contact with it. This contact time depends on the flow rate of the C0 2 .
- the solvent is placed in a flask and the gas flow is passed through a capillary.
- the second problem arises especially when collecting for a longer period of time by evaporating the solvent. Feeding the solvent with the amount per time, which also evaporates per time, can keep the amount of solvent in the receiver constant, but does not prevent the losses due to evaporation of the sample itself. A reduction in the template temperature counteracts this. However, the temperature must not be reduced to such an extent that the capillary is blocked by freezing of the C0 2 (for example by acetone / dry ice).
- Nozzle temperature during extraction 90 ° C
- Nozzle temperature for the rinse step 70 ° C
- the traps with solvent (10 ml of n-heptane in the receiver) are cooled to 0 ° C in an ice bath.
- an external pump is used to add n-heptane.
- the n-heptane is fed directly to the outlet of the nozzle and flows through the capillary into the template.
- the flow rate is chosen so that approximately the amount of n-heptane that evaporates per time is balanced.
- the gas flow escapes through small channels on the plug of the flask (50 ml volumetric flask) after it has passed the template. With this arrangement, only 35% of the 18: 1 EA is recovered.
- the trap technique with RP-C18 material leads particularly well to a quantitative recovery. This is used in the following for all further optimization attempts. It can also be scaled up if the amount of sorbent is increased. The separate rinsing step is disadvantageous. On a preparative, technical scale, cyclone separators can be used that collect the pure product without working with solvents.
- sample preparation includes various options for how and where the sample is added to the system.
- the model substance triolein is liquid at room temperature.
- the reactor is about 4/5 filled with catalyst.
- Triolein is then weighed directly onto the catalyst. Then you fill the reactor with catalyst.
- the sample is thus in direct contact with the catalyst and is surrounded by it.
- the direction of the fluid flow is chosen so that 4/5 of the catalyst is available as a "reaction path", ie it is extracted from the end where the sample was placed through the reactor. This technique is used for all optimizations with triolein.
- solid samples are weighed in with the catalyst and ground in a mortar in order to produce the largest possible contact area.
- Acidic aluminum oxide with activity level Super I (name: Alox A Sl) was used with the following weights:
- the modifier content affects the extraction on the one hand and the reaction on the other. For the latter, the higher the proportion of the reactant ethanol, the higher the reaction yield that can be achieved. On the other hand, it should be noted that with increasing modifier content, the extraction yield increases not only for 18: 1 -EE, but also for the educt triolein. Therefore, the addition of ethanol in the fluid must be optimized in addition to the pre-loading of the catalyst with ethanol.
- the SF-REC for triolein is carried out under the same conditions, with 1% and 2% ethanol being added as a modifier in the first case. A faster reaction and a higher yield are found for the higher ethanol content.
- the increase in the ethanol content is limited by the increased extraction of triolein. It must be prevented that the educt is removed before it can react.
- the density and thus the solvent strength can be adjusted via the pressure and the temperature of the fluid. This is crucial for the extraction yield. It can be used to control the selectivity of the extraction between the ethyl ester product and the triglyceride starting material. As shown in the previous chapter, the modifier content also affects this selectivity. Overall, the SF-REC condition must therefore be selected so that, with the highest possible modifier content, the density is so low that only the ethyl ester is selectively extracted from the reaction space. However, the temperature, like the ethanol content, also has an effect on the reaction rate. If the temperature is increased at the same density, the yield per time increases, as shown in FIG. 4: In principle, the temperature in the given system can be increased by up to 150 ° C.
- Temperature and modifier content have a direct influence on the reaction yield, i.e. the turnover rate of the transesterification. They should both be as high as possible in order to achieve a quick implementation.
- the extraction of the product from the reaction space should be as selective as possible with respect to the starting material, i.e. the solvent strength of the fluid must be sufficient to achieve extraction of the ethyl ester which is as complete as possible but extraction of the triglyceride as low as possible. Accordingly, the following strategy was used for optimization: By lowering the pressure while simultaneously increasing the temperature and the modifier content, the extraction can be sufficiently selective, but the conversion rate can be correspondingly high. The optimized conditions are listed below:
- Nozzle temperature during extraction 90 ° C
- a lecithin with two oleic acids dioleylphosphatidylcholine was chosen as the second model substance. This was implemented under the conditions optimized for triolein. The reaction kinetics can be seen in FIG. 5: If one compares the kinetics with that for the transesterification of the triolein, it is found that the reaction proceeds more slowly under the same conditions for the phospholipid (FIG. 5).
- the solvent strength of the fluid can be adjusted. This allows selective extraction of the product ethyl ester.
- the reaction rate is increased by increasing the reaction temperature and the ethanol content, while at the same time keeping the density so low that the selectivity of the extraction / chromatography is retained.
- the SF-REC conditions can thus be optimized so that a quantitative transesterification from triolein to oleic acid ethyl ester can take place.
- Example 13 Application to a biological source / matrix: dry biomass GLA strain:
- a freeze-dried (lyophilized) powder which is obtained from the fermentation of microorganisms, is referred to as dry organic matter.
- the first dry biomass comes from a so-called GLA strain. This contains a particularly large amount of ⁇ -linolenic acid (GLA), which is usually bound in phosphatides.
- GLA ⁇ -linolenic acid
- the n-heptane solution obtained from the second step was analyzed by GC-MS to identify the fatty acid ethyl esters present.
- the chromatogram and a section are shown in FIG. 6.
- fatty acid ethyl esters in addition to the GLA-EE, a number of other fatty acid ethyl esters can be found in FIG. 6, in particular oleic and linoleic acid ethyl esters (18: 1-EE and 18: 2-EE), but also C14, C16 and C20 fatty acids.
- the aqueous suspension of the biomass (without lyophilization) is called the bio-moist mass. If these are converted directly in an SF-REC, the excess water does not cause the lipids to be transesterified into the ethyl esters, but instead hydrolyses them to the free acids. This is confirmed by the analysis using GC-MS (FIG. 7).
- the SF-REC was carried out under the following conditions: Flow: 1.0 ml / min
- the lyophilized biomass of the DHA strain mainly contains docosapentaenoic acid (DPA 22: 5).
- the total lipidanteii of the dry biomass is about 40% by weight. This in turn contains 40% of the fatty acids DHA.
- the lipid content is almost exclusively in the form of triglycerides.
- the dry biomass was ground in a mortar with ten times the amount of acidic aluminum oxide activity Super I, which was coated with 15% ethanol, and filled into the reactor. The SF-REC was carried out under the following conditions:
- the amount of DHA-EE was determined using a DHA standard (90%, KD-PHARMA, Bexbach).
- both the lyophilized and an aqueous suspension of the biomass could be implemented.
- the ethyl esters are formed in a transesterification reaction with ethanol
- the free acids are formed by hydrolysis.
- the fatty acids as ethyl esters could also be obtained from the dry biomass of the DHA strain, which mainly contains docosahexaenoic and docosapentaenoic acid.
- Example 16 Application of SF-REC - extraction of valuable biogenic substances from algae: The aim of the experiments carried out here is to examine the possibility of extracting valuable substances from algae using various extraction methods. The main focus is on polyunsaturated fatty acids, which are found in algae in quite high concentrations. Arachidonic acid (5,8,11, 14-eicosatetraenoic acid; ERA) and eicosapentaenoic acid (EPA) are particularly worth mentioning here. In addition, unsaturated fatty acids with 18 carbon atoms such as gamma-linolenic acid (GLA) are of particular importance.
- GLA gamma-linolenic acid
- algae In addition to these substances, depending on the strain, algae also contain a number of dyes such as chlorophyll, xanthophylls (carotenoids), but also some specific ones, such as the class of phycobiliproteins. In red algae (Rhodophytha) in particular the phycoerythrins are found.
- SFE with pure C0 2 represents the first step in the extraction of fatty acids from algae.
- Polar compounds can then be extracted in the second step by using ethanol as a modifier.
- An extraction of polar compounds, especially those that show good water solubility, cannot be done with CO 2 , so that these can then be extracted with water in a third step.
- the chlorophyll With increasing pressure and also by using 10% ethanol as a modifier, the chlorophyll can be removed from the algae material almost completely after several hours of extraction.
- Example 20 Extraction in the presence of an adsorbent / in-situ transesterification by SF-REC:
- the algae were now ground with an adsorbent (aluminum oxide) instead of sea sand and extracted under the same conditions as above. It is found that the fractions which were obtained without a modifier are not green, but only slightly yellow.
- the UV spectrum up to 600 nm chlorophyll could be recorded using a diode array detector.
- the yellow fraction therefore contains no chlorophyll.
- the peak at 3.1 min was identified as yellow dyes by moving a thin-layer plate past the outlet of the SFC at a defined speed. A bluish fluorescent spot was found on it at 3.1 min under UV light. It is thus possible to transesterify the valuable fatty acids from the algae in situ and to isolate them as ethyl esters without chlorophyll being extracted.
- the direct extraction of the algae with ethanol provides green solutions.
- the chlorophyll can therefore be removed from the algae with ethanol.
- Example 22 Extraction with water A purple solution can be obtained from water both from the algae “pre-cleaned” with SFE and from the untreated algae. When excited with UV light (366 nm), this solution shows orange fluorescence. The purple dye can thus be separated from the fats and chlorophyll in an aqueous step.
- the aluminum oxide initially used can be separated from the algae residues in water by sedimentation. It shows a mint green color, which suggests adsorbed chlorophyll.
- the color of the yellow solution can be explained in the spectrum by the flank, which extends from the UV range to 500 nm. An absorption maximum can be seen in the UV range at 282.4 nm. The cause of the green coloration of the chlorophyll solution is the absorption maximum at 662.6 nm. Numerous maxima of lower height can be found in the range of visible light. The highest maximum is 410 nm. The purple solution absorbs most strongly in the visible range at 545.5 nm (according to the literature, the algal dye B-phycoerythrin has an absorption maximum at 546 nm). In the UV range there is a maximum at 272.2 nm, which may be due to light scattering from the slightly cloudy solution. The examples show that extraction and separation of the desired substances, for example from the given algae, is possible.
- the valuable polyunsaturated fatty acids can be obtained as ethyl esters. They are extracted along with yellow dyes, which are probably xanthophylls. These can be processed in a further processing step e.g. can be separated using preparative SFC.
- the disruptive chlorophyll can either be removed from the matrix using SFE with more than 10% ethanol as a modifier and higher pressure (higher density) or by direct extraction with ethanol. The purple dye of the algae is not extracted.
- the advantages of the methods used lie in the fact that any contact with toxic solvents can be avoided, so that the substances obtained are available for pharmaceutical or food technology use.
- the conversion of the triglyceride triolein to oleic acid ethyl ester served as a model example for the transesterification using ethanol on acidic aluminum oxide.
- the SF-REC was optimized for this example in terms of equipment conditions and process control. An almost quantitative transesterification was achieved with a yield of 93%.
- the SF-REC could also be carried out for the model substance of the phosphatide dioleylphosphatidylchoii ⁇ .
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Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP99938281A EP1102859A1 (de) | 1998-07-22 | 1999-07-17 | VERFAHREN ZUR PRÄPARATIVEN GEWINNUNG VON FETTSÄUREN AUS BIOMASSE DURCH $i(IN- SITU)-EXTRAKTION-REAKTION-CHROMATOGRAPHIE MIT VERDICHTETEN GASEN |
JP2000561341A JP2002521030A (ja) | 1998-07-22 | 1999-07-17 | 圧縮ガスを使用する同一装置における抽出、反応及びクロマトグラフィーによるバイオマスからの脂肪酸の生産規模製造方法 |
US09/744,142 US6350890B1 (en) | 1998-07-22 | 1999-07-17 | Method for obtaining fatty acids from biomass by combined in/situ extraction, reaction and chromatography using compressed gases |
NO20010276A NO20010276L (no) | 1998-07-22 | 2001-01-17 | FremgangsmÕte for utvinning av fettsyrer fra biomasse ved insitu ekstraksjon, reaksjon og kromatografi med fortettede gasser |
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DE19832784A DE19832784A1 (de) | 1998-07-22 | 1998-07-22 | Verfahren zur präparativen Gewinnung von Fettsäuren aus Biomasse durch in situ-Extraktion-Reaktion-Chromatographie mit verdichteten Gasen |
DE19906474.1 | 1999-02-12 | ||
DE19832784.6 | 1999-02-12 | ||
DE19906474 | 1999-02-12 |
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WO2000005395A1 true WO2000005395A1 (de) | 2000-02-03 |
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US (1) | US6350890B1 (de) |
EP (1) | EP1102859A1 (de) |
JP (1) | JP2002521030A (de) |
NO (1) | NO20010276L (de) |
WO (1) | WO2000005395A1 (de) |
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DE10159987A1 (de) * | 2001-12-06 | 2003-06-26 | Nutrinova Gmbh | Verfahren zur Aufreinigung von Glycerolether enthaltenden Mischungen |
US9023616B2 (en) | 2006-08-01 | 2015-05-05 | Dsm Nutritional Products Ag | Oil producing microbes and method of modification thereof |
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US10537840B2 (en) | 2017-07-31 | 2020-01-21 | Vorsana Inc. | Radial counterflow separation filter with focused exhaust |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61261398A (ja) * | 1985-05-15 | 1986-11-19 | 株式会社日立製作所 | 魚油精製方法 |
US4898673A (en) * | 1987-07-11 | 1990-02-06 | Vitamins, Inc. | Dynamic supercritical fluid extraction system |
-
1999
- 1999-07-17 EP EP99938281A patent/EP1102859A1/de not_active Withdrawn
- 1999-07-17 WO PCT/EP1999/005107 patent/WO2000005395A1/de not_active Application Discontinuation
- 1999-07-17 JP JP2000561341A patent/JP2002521030A/ja active Pending
- 1999-07-17 US US09/744,142 patent/US6350890B1/en not_active Expired - Lifetime
-
2001
- 2001-01-17 NO NO20010276A patent/NO20010276L/no unknown
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61261398A (ja) * | 1985-05-15 | 1986-11-19 | 株式会社日立製作所 | 魚油精製方法 |
US4898673A (en) * | 1987-07-11 | 1990-02-06 | Vitamins, Inc. | Dynamic supercritical fluid extraction system |
Non-Patent Citations (4)
Title |
---|
CHEMICAL ABSTRACTS, vol. 106, no. 20, 18 May 1987, Columbus, Ohio, US; abstract no. 158274, MATSUZAKI, HARUMI ET AL: "Purification of fish oil" XP002123462 * |
CUMMINS, M. T. AND WELLS, R. J.: "In situ derivatisation and extraction of volatile fatty acids entrapped on anion exchange resin from aqueous solutions.", JOURNAL OF CHROMATOGRAPHY B: BIOMEDICAL APPLICATIONS, vol. 694, 1997, pages 11 - 19, XP004081995 * |
KING, J. W.: "Analytical-process supercritical fluid extraction: a synergistic combination for solving analytical and laboratory scale problems.", TRENDS IN ANALYTICAL CHEMISTRY, vol. 14, no. 10, 1995, pages 474 - 481, XP004034803 * |
WENCLAWIAK B. W. ET AL.: "In situ transesterification of the natural pyrethrins to methyl esters by heterogenic catalysis using a supercritical fluid extraction system and detection by GC-MS.", JOURNAL OF CHROMATOGRAPHY A, vol. 785, 1997, pages 263 - 267, XP004097570 * |
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EP1024199A3 (de) * | 1999-01-27 | 2002-03-06 | Celanese Ventures GmbH | Gewinnung von Gamma-Linolensäure aus Protozoen der Gattung Colpidium |
EP1024199A2 (de) * | 1999-01-27 | 2000-08-02 | Axiva GmbH | Gewinnung von Gamma-Linolensäure aus Protozoen der Gattung Colpidium |
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Also Published As
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JP2002521030A (ja) | 2002-07-16 |
US6350890B1 (en) | 2002-02-26 |
EP1102859A1 (de) | 2001-05-30 |
NO20010276D0 (no) | 2001-01-17 |
NO20010276L (no) | 2001-03-20 |
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