JPS60241908A - Method for producing purified components from multi-component raw materials - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a method for separating and extracting a desired component from a multi-component raw material containing a large number of components in addition to the desired component such as natural products, extracts, and reaction solutions. The present invention relates to a method for producing purified components from multi-component raw materials. That is, in this invention, a multi-component raw material is supplied to a separation tube filled with a stationary phase packing, and pressurized carbon dioxide is passed through the separation tube as a mobile phase fluid to separate the target component from other components. The present invention relates to a method for producing a purified component by extracting a target component.

The pressurized carbon dioxide in the present invention refers to carbon dioxide that is pressurized to make a fluid that can be used as a mobile phase, and carbon dioxide in a liquid or supercritical state is used. In other words, if carbon dioxide is used at a temperature and pressure above the critical point (31.1°C75.2~/m), it will not become supercritical, but at a temperature lower than the critical temperature and at a pressure higher than the boiling curve. When carbon dioxide is used, it becomes a liquid.

In addition, stationary phase packing refers to a substance that adsorbs, releases, and distributes the target component with moderate force in the mobile phase fluid, and separates the target component from other components, such as activated carbon, calcium carbonate, etc. , silica gels such as quartz, chemically bonded silica gel, diatomaceous earth materials such as sintered diatomaceous earth, porous polymers, and polymer compounds such as polar or non-polar exchange resins that do not dissolve or react with pressurized carbon dioxide. Substances are used. Note that the filler used has an appropriate particle size so that the mobile phase fluid flows through the gap when the separation tube is filled.

In addition, the multicomponent raw material in the present invention refers to a mixed substance containing many components such as different compounds, isomers, homologs, unreacted substances, impurities, solvents, and reaction promoters in addition to the target component. It refers to natural products, crude products obtained by concentrating or extracting natural products, or reaction liquids after the reaction has been completed.

Distillation methods, solvent extraction methods, crystallization methods, and the like have been used to extract target components from multi-component raw materials in which many components coexist and produce purified components.

To obtain multi-component raw materials and N-made components by distillation, many methods are used, such as atmospheric distillation, reduced pressure or vacuum distillation, molecular M'fd, multi-stage distillation, fractional distillation, and steam distillation.

That is, for example, citrus essential oil can be distilled under reduced pressure or vacuum, separated into a low-boiling point portion and a high-boiling point portion, and terpene-free essential oil of citrus fruits can be produced by obtaining a low-boiling point portion that contains relatively few terpenes. It is being said.

Furthermore, molecular distillation of fats and oils containing many triglycerides has been carried out to produce fats and oils with a specific fatty acid composition.

However, for example, fish oil has a high content of cyeicosapentaenoic acid and docosahexaenoic acid.
−) Extracting the desired component from a multi-component raw material containing components with similar properties and similar boiling points, such as when concentrating α-tocopherol, which has greater physiological activity than natural tocopherol mixed with copherol. was difficult and what was obtained was expensive.

Furthermore, the distillation method cannot be used for high-boiling polymer compounds because the target component must evaporate under the processing conditions, and it is not suitable for thermally unstable substances because it is processed by applying temperature.

The crystallization method is used to produce refined ingredients, such as crystallizing sugar from a sugar solution, crystallizing menthol from mint oil, and separating fats and oils using fractional crystallization to obtain fats and oils with a specific melting point. It's being used.

However, in general, to generate crystals, it is necessary to concentrate a solution, add seed crystals, cool it to a certain temperature, and leave it for a long time, which takes time. Moreover, uncrystallized components remain in the solution after the crystals are separated, and the recovery rate is not very good. Furthermore, it cannot be used for compounds that are difficult to crystallize or non-crystalline compounds, such as high-molecular compounds and compounds with high solubility.

Extraction methods are widely used to obtain desired components from multi-component raw materials containing insoluble components, such as tea, coffee, Monascus pigment, paprika pigment, etc., by dissolving them in water or other solvents. There is.

In addition, extraction is also carried out using a solvent that is immiscible with liquid or solution-like multi-component raw materials, such as when extracting terpene-free citrus essential oil using a dialcohol aqueous solution. ing.

Furthermore, when a liquid multi-component raw material and a solvent mix with each other, such as when extracting an aqueous solution containing salts with water, dialysis, reverse osmosis, Electrodialysis and other methods may also be used.

However, no matter which extraction method is used, the extract usually contains a large amount of solvent, so it is necessary to concentrate and separate the desired components from the extract by distillation, crystallization, or other methods.

Moreover, when an extract containing a large number of components is obtained, such as an extract from a natural product, further separation and purification are required to obtain the desired components, which is time-consuming.

Furthermore, when processing with organic solvents such as extraction with organic solvents or recrystallization, solvents may remain in the resulting purified components, which may make them unusable for human consumption such as food. In addition, organic solvents may even be dangerous, as care must be taken against fire and explosion.

Moreover, in either method, the target components are separated and purified from components with very similar properties (such as homologues and isomers) and multi-component raw materials, and impurities present in extremely small amounts are removed. It is difficult to remove.

This invention enables the production of purified components easily without the time and effort required by conventional methods, and also allows for the production of homologs,
The purpose of the present invention is to provide a new manufacturing method that can produce purified components even from multi-component raw materials that are difficult to separate, such as isomers.

That is, according to the present invention, when pressurized carbon dioxide in a liquid or supercritical state is flowed into a separation tube as a mobile phase fluid and a multi-component raw material is supplied to the starting end of the separation tube, each component in the multi-component raw material is The components dissolve in carbon dioxide and move through the separation tube to the end of the tube. At this time, each component moving within the separation tube moves while repeating adsorption onto the stationary phase packing filled in the separation tube and release into the mobile phase fluid consisting of carbon dioxide. Moreover, since the adsorption power to the stationary phase packing and the solubility in the mobile phase fluid differ depending on each component, the time required for the components to be distributed, to pass through the separation tube, and to flow out from the terminal end differs. Therefore, when the target component comes out from the end of the separation tube, the purified target component can be obtained by collecting that portion.

In addition, a multi-component raw material is used, which is a mixture of components that have weak adsorption to the stationary phase packing and high solubility in the mobile phase fluid, and components that have strong adsorption to the stationary phase packing and low solubility in the mobile phase fluid. In this case, only the components that are weakly adsorbed to the stationary phase packing may flow out of the separation tube, and the components that are strongly adsorbed to the stationary phase packing are adsorbed by the stationary phase packing and may not flow out of the separation tube. Therefore, in such multi-component raw materials, purified target components can be obtained by extracting the components dissolved in the mobile phase fluid or the components adsorbed on the stationary phase packing, depending on the purpose. .

Furthermore, carbon dioxide can become a supercritical liquid by changing the pressure and temperature, so the solubility of each component in the mobile phase liquid and adsorption to the stationary phase packing vary depending on the state. Change. By taking advantage of this property and adjusting the pressure and temperature appropriately, we were able to separate difficult-to-separate homologs and isomers.

In addition, as a method for separating and purifying the desired components from multi-component raw materials using pressurized carbon dioxide as an extraction fluid,
Extraction methods using liquid or supercritical carbon dioxide have recently been announced. However, in this case as well, an extract containing all the components extracted by pressurized carbon dioxide is obtained in the same way as in the conventional extraction method, and as in the present invention, only the specific components of interest are separated and purified. It was impossible to do so.

In addition, the stationary phase packing packed in the separation tube is selected as appropriate depending on the target components to be separated, and if necessary, water, alcohols such as ethyl alcohol, ketones such as acetone, aldehydes such as acetaldehyde, dimethyl ether, etc. One or more of ethers such as ethyl acetate, esters such as ethyl acetate, and hydrocarbons such as benzene and hexane are used as the entrainer.

When using an entrenner, it is carried out by a known method, such as mixing the stationary phase packing and the entrenner and then filling the mixture into a separation tube, or adding it to pressurized carbon dioxide as a mobile phase fluid. The ratio of the stationary phase packing or mobile phase fluid to the entrainer is determined as appropriate, but when added to the stationary phase packing, it is usually about 1 to 60 parts per 100 parts.
Also, when added to the mobile phase fluid, approximately 0.0% per 100 parts thereof.
Preferably, 0.01 to 30 parts of entrenor are used.

To supply a multi-component raw material to the separation tube, pressurized carbon dioxide is injected into the beginning of the separation tube and then sent to the separation tube, or an injection tube is inserted into the flow of carbon dioxide and an appropriate amount is injected from there. Inject and spray multi-component raw materials to dissolve them in a fluid.
This is done by methods such as dispersion.

In addition, a bypass is provided in the flow path that supplies pressurized carbon dioxide to the separation tube, and an appropriate liter of multicomponent raw material is put into the bypass section, and carbon dioxide passes through the bypass by switching the cock, etc. It can also be carried out by allowing it to go to a separate tube. In particular, this method is suitable when using a solid multi-component raw material and supplying the raw material while extracting components that dissolve in pressurized carbon dioxide.

To recover the target purified component separated from the multi-component raw material, the mobile phase fluid flowing out from the separation tube is fractionated at a constant rate of 1.0 cm, and the fraction containing the target component is taken out. This is done by installing a sensor at the end of the separation tube that measures absorbance, thermal conductivity, specific heat, etc. to detect the component flowing out, and switching the flow path of the mobile phase fluid when the target component is detected. It is also possible to recover it by

Note that when other components are adsorbed by the stationary phase packing and only the target component flows out from the separation tube, the target purified component can also be obtained by collecting all of the mobile phase fluid that flows out.

The pressure of the separated and collected mobile phase fluid is lowered to normal pressure to volatilize carbon dioxide, leaving behind the purified target component.

The purified components obtained in this way do not have to worry about residual solvents, unlike extraction methods or crystallization methods, and since they are processed in an oxygen-free condition and at relatively low temperatures, there is no risk of deterioration or rancidity. do not have. Therefore, it can be safely used to purify ingredients that can be taken into the body, such as pharmaceuticals and foodstuffs, and since no organic solvent is used, it can be carried out safely with almost no fear of fire or explosion.

In addition, the purified components that flow out from the separation tube together with the mobile phase fluid can be collected by connecting an adsorption tube filled with an adsorbent made of NT acid components such as activated carbon to the separation tube, and adsorbing and recovering the purified components that flow out from the separation tube. It is also possible to do so.

Further, the purified components separated by adsorption onto the stationary phase packing can be recovered by flowing an eluent with which the purified components are eluted through a separation tube. Further, the purified components recovered using the adsorption tube are similarly taken out by flowing the eluent through the adsorption tube. The eluent used at this time can be arbitrarily selected depending on the purpose, such as pressurized fluids of carbon dioxide or other gases with different conditions such as temperature and pressure from those used during separation, or liquid fluids such as organic solvents and solutions. do.

Next, an example will be explained.

Example 1 Silica gel (manufactured by Fuji Devin ■, Cylobia 20μ) as stationary phase packing 21! After filling the separation tube with hexane, carbon dioxide gas was passed into the separation tube while keeping the temperature at about 60° C. to completely evaporate and remove hexane, thereby preparing a separation tube.

Next, sardine oil 502, which was composed of 19.3% of the fatty acids consisting of eicosapentaenoic acid and 9.8% of docosahexaenoic acid and had a slightly yellowish color and had a so-called fish oil component, was supplied to the separation tube.

Next, the temperature of the separation tube was maintained at 36°C and 3-/3
-oo4. Carbon dioxide pressurized to 'i]00ri/10i
The mobile phase fluid flowing out from the separation tube was fractionated, the carbon dioxide was evaporated, and the contents of eicosapentaenoic acid and docosahexaenoic acid that remained were measured as shown in the table. became.

Note that 98% of the pressurized carbon dioxide is added with ethyl alcohol as an entrainer in advance.
Additions were used at a rate of 2 volumes per volume.

The unsaturated fatty acid content of each fraction of sardine oil separation is shown in Table 1. Therefore, the amount of carbon dioxide effluent is classified as 1.2 to 2.0 (
By taking out fraction 46) and reducing the pressure while warming it to normal pressure, a colorless and transparent oil with a high content of eicosapentaenoic acid and docosahexaenoic acid and no fish oil ingredients was obtained. Obtained.

Example 2 Non-polar porous polymer resin (manufactured by Mitsubishi Kasei ■, HP-20)
5. Ol! was poured into a separation tube together with ethyl alcohol as a stationary phase packing, and the mixture was heated to about 70° C. and carbon dioxide gas was passed through it to completely remove the ethyl alcohol, thereby preparing a separation tube.

Next, 200 y of fish oil in which 17.2% of the constituent fatty acids are eicosapencitric acid and 8.3% are docosahexaenoic acid.
- is injected from the starting end of the separation tube, and the temperature of the separation tube is set to 38°C.
300 while keeping K? /m of carbon dioxide pressurized to 2
The carbon dioxide flowing out from the end of the separation tube was fractionated and collected.

A sensor was installed at the end of the separation tube to measure the absorbance of the outflowing carbon dioxide over time, as shown in Figure 1.

The pressurized carbon dioxide gas (corresponding to the category E in Figure 1) that flowed out for 20 minutes from 40 minutes after the start of flow was removed to obtain a colorless, odorless, and transparent oil of 25.6%.

The eicosapentaenoic acid content of this refined oil was 29.6% in terms of constituent fatty acid ratio.

Further, docosahexaenoic acid was 17.3%.

Example 3゜Chemically bonded C-18 silica gel (Merck & Co. gLichr
oprep RP-18) 2.5/ was suspended in ethyl alcohol and filled into a separation tube as a stationary phase packing,
A separation tube is prepared by heating the tube to about 70°C and flowing carbon dioxide gas to completely remove ethyl alcohol inside the separation tube.

The separated tube was then placed in a thermostat at 38°C and 6 (1) natural tocopherols were injected.

When this natural tocopherol is analyzed by high-performance liquid chromatography, it becomes as shown in Figure 2, α-1β-1γ-
It is composed of a multi-component mixture of 1δ-tocopherol, and α is 26.4%.

Next, carbon dioxide pressurized to 360 to 9/m was flowed through the separation tube, and the absorbance of the carbon dioxide flowing out at the end of the separation tube was measured over time. The results were as shown in Figure 3.

The fraction corresponding to FIG. 3A of the carbon dioxide flowing out from the separation tube was taken out, and purified α-tocopherol was obtained by removing carbon dioxide.

The analysis results of this α-tocopherol by high performance liquid chromatography were as shown in FIG. 4, and the purity of α-toco7erol was 92.8%.

Example 4 A separation tube was filled with 21 silica gel (Silopure, manufactured by Fuji Davison Chemical Co., Ltd.) which had been suspended in ethyl alcohol in advance, and carbon dioxide gas was bubbled through the tube at 70°C to completely remove the ethyl alcohol. Then rotate the separation tube at 36°
Lemon oil from 1 to 1 was injected as C. The lemon oil before this treatment contains a large amount of terpenes, as shown in its gas chromatography in FIG.

Next, the carbon dioxide, which had been pressurized by adding ethyl alcohol 17- to 2 volumes per 98 volumes of liquid carbon dioxide at 20°C, was kept at 36°C and placed in a separation tube. / m i H flow rate. In addition, carbon dioxide flowing out from the separation tube was passed through an adsorption tube filled with activated carbon as an adsorbent.

After flowing pressurized carbon dioxide for about 35 minutes, stop the flow of carbon dioxide and pour ethyl alcohol into the separation tube.
The terpene-free lemon oil adsorbed on the stationary phase packing in the separation tube was eluted and recovered.

This terpene-free lemon oil contained very few terpenes, as seen in the gas chromatography shown in Figure 6.

In addition, terpenes adsorbed in the adsorption tube were eluted and recovered by flowing ethyl alcohol through the adsorption tube.

Example 5 Chemically bonded silica gel C-8 (Lichroprep C
-8,25-407JTIMerck) 3. OI! A slurry in which ethyl alcohol was suspended in ethyl alcohol was filled into a separation tube as a stationary phase packing, kept at 70°C, and ir-carbon dioxide gas was bubbled through to completely remove ethyl alcohol, thereby preparing a separation tube.

Commercially available Monascus pigment 200y, which is a crude extract obtained by extracting Monascus mold with 85% ethyl alcohol and removing the solvent, was injected into a separation tube, and carbon dioxide pressurized to 200Kg/cII at 38°C was added at 110y/cII. When the absorbance of carbon dioxide flowing out from the end of the separation tube was measured over time when flowing at a flow rate of rn i 11, the results were as shown in FIG. 7.

Monascus pigment 16 purified by fractionating carbon dioxide in the category corresponding to AK in Figure 7 and removing carbon dioxide.
Got 7y.

In addition, fl! The absorption curves of the Monascus dye before I production and after purification are A and B in FIG. 8, respectively, and the absorbance at 485 nm increased by about 20% due to purification.

Example 6 After extracting the peel of paprika (Capsicum annum L) with 90% ethyl alcohol, the solvent was removed to obtain oleoresin. 200 commercially available paprika pigments, which are paprika crude extracts, were extracted in the same manner as in Example 5. The mixture was injected into the prepared separation tube, and carbon dioxide kept at 38°C and pressurized to 13QKg/cm was flowed therein. The flow rate of carbon dioxide at this time was 100 P/min.

In addition, the absorbance of carbon dioxide flowing out from the end of the separation tube was measured over time, and the results were as shown in Figure 9.

Therefore, 1982 purified paprika pigments were obtained by fractionating the carbon dioxide corresponding to part A in FIG. 9 and removing the carbon dioxide.

The absorption curves of paprika pigment before purification and paprika pigment after purification are as shown in curve A and curve B in Figure 10.
The absorbance of the purified paprika pigment at 464 nm was 42% higher than that of the crudely extracted paprika pigment before purification.

[Brief explanation of drawings]

Figure 1: Changes over time in absorbance of carbon dioxide flowing out of Example 2 (fish oil), where the horizontal axis is time (minutes) and the vertical axis is absorbance. Figure 2: High performance liquid chromatography of tocopherol before purification in Example 3, where the horizontal axis is time and the vertical axis is absorbance. Figure 3: Absorption curve of carbon dioxide flowing out of Example 3 (tocopherol), where the horizontal axis is time and the vertical axis is absorbance. Figure 4: High performance liquid chromatography of purified α-tocopherol in Example 3, where the horizontal axis is time (minutes) and the vertical axis is absorbance. Figure 5: Gas chromatography of lemon oil before purification in Example 4. Figure 6: Gas chromatography of purified terpene-free lemon oil of Example 4. FIG. 7: Changes over time in absorbance of carbon dioxide flowing out of Example 5 (Monascus dye), where the horizontal axis is time (minutes) and the vertical axis is absorbance. FIG. 8: Absorption curve of the Monascus dye of Example 5 (A before purification, B after purification), where the horizontal axis is wavelength (nm) and the vertical axis is absorbance. Figure 9: Time-dependent change in absorbance of carbon dioxide flowing out of Example 6 (paprika dye), where the horizontal axis is time and the vertical axis is absorbance. Figure 10: Absorption curve of paprika dye of Example 6 (A
(before purification, B after purification), the horizontal axis is wavelength (nm), and the vertical axis is absorbance. -l- 350 450 8) Onm Wavelength → Wavelength □ Procedural amendment (voluntary) 1. Indication of the case >7- tar 'f' ρZ Patent application filed on May 15, 1980 2 Name of the invention Multi-component system Method for producing purified components from raw materials 3, Relationship with the case of the person making the amendment Patent applicant address: 5-33-1-4 Shiba, Minato-ku, Tokyo Subject of amendment 5, Contents of amendment (11th Specification No. 1) "Made by Fuji Devison ■" on page 13, lines 11-12 is corrected to "manufactured by Fuji Devison Chemical Co., Ltd." (2) "...Passed through an adsorption tube" on page 18, line 4 of the specification is changed to "... ... through an adsorption tube." (3) Insert the following Example 7 between lines 16 and 17 on page 20 of the specification. "Example 7 Suspended in ethyl alcohol. Fill the separation tube with approximately 21! of silica gel (Silopia, manufactured by Fuji Davison Chemical Co., Ltd.),
Next, the tube was maintained at 70 to 80.degree. C. and carbon dioxide gas was passed through it to completely remove ethyl alcohol, thereby preparing a separation tube. While maintaining this separation tube at 20°C, IKg of lemon oil was injected, then 2 volumes of ethyl alcohol was added per 98 volumes of liquid carbon dioxide, and the carbon dioxide was pressurized to 90~/cIIt to 100 f! / min at a flow rate of approximately 4
It was run for 5 minutes. Note that the carbon dioxide flowing out of the separation tube was subsequently passed through an adsorption tube filled with activated carbon as an adsorbent. Next, ethyl alcohol was poured into the separation tube instead of the pressurized carbon dioxide to elute and collect the terpene-less oil adsorbed on the stationary phase packing. Ethyl alcohol was also poured into the adsorption tube to obtain terpenes. (4) Amend Figure 1 as shown in the attached drawing.

Claims (1)

[Scope of Claims] 1. Purpose of supplying multi-component raw materials to a separation tube filled with a stationary phase packing material and flowing pressurized carbon dioxide into the separation tube as a mobile phase fluid. 1. A method for producing lnI components from multi-component raw materials, the method comprising separating target components and recovering the separated target components. 2. A method for producing a purified component from a multi-component raw material according to claim 1, wherein the target component is recovered by flowing out the target component together with carbon dioxide through a separation tube. 3. A method for producing a purified component from a multi-component raw material according to claim 1, wherein the target component is recovered by adsorbing the target component onto a stationary phase packing. 4. The method for producing purified components from multicomponent raw materials according to claim 1, wherein the pressurized carbon dioxide is carbon dioxide in a supercritical state. 5. The method for producing purified components from multicomponent raw materials according to claim 1, wherein the pressurized carbon dioxide is carbon dioxide in a liquid state.