JPH04255799A - Extraction of volatile effective component - Google Patents

Abstract

PURPOSE:To extract and separate volatile effective components from a material to be extracted in high yield and efficiently without changing properties of the volatile effective components. CONSTITUTION:In extracting volatile effective component such as fragrant component from a material to be extracted, super critical carbon dioxide or carbon dioxide close to a super critical state at 20-80 deg.C under 70-400kgf/cm<2> gauge pressure is used as an extractant.

Description

[Detailed description of the invention]

0001

[Field of Industrial Application] The present invention relates to a method for extracting volatile active ingredients, and more specifically, extracts volatile active ingredients from a material to be extracted, such as flowers, buds, fruits, seeds, foliage, bark, etc. of plants.
This invention relates to a method for extracting and separating volatile active ingredients such as aroma ingredients contained in stems, rhizomes, etc. using supercritical carbon dioxide or carbon dioxide in a near supercritical state.

0002

[Prior art] Flowers, buds, fruits, seeds, branches and leaves of plants,
The bark, stem, rhizome, or cultured cells thereof contain active ingredients such as pharmaceuticals, foods, and fragrances. These active ingredients are volatile and often contained in trace amounts. Conventionally, these volatile active ingredients have been extracted by extraction methods such as steam distillation and solvent extraction.

[0003] However, when volatile active ingredients are extracted by steam distillation, the distillation is carried out at high temperatures, so the heat-labile ingredients change in quality during distillation, and the active ingredients originally contained in plants are There is a problem in that foreign components are extracted. In addition, water-soluble substances dissolve in the aqueous layer, resulting in a decrease in yield, or when recovering components dissolved in the aqueous layer, a recovery operation is required, which is complicated and evaporates during the recovery operation. There is a problem in that the yield decreases due to the escape of active ingredients. Further, after steam distillation, it is necessary to separate an oil layer and an aqueous layer, but this separation process also causes the problem that volatile active ingredients escape, resulting in a decrease in yield.

On the other hand, when volatile active ingredients are extracted using an organic solvent, the volatile active ingredients may be deteriorated by the organic solvent. Furthermore, after extraction, it is necessary to separate the volatile active ingredient and the organic solvent, but this is usually done by adding thermal energy to the boiling point difference between the volatile active ingredient and the organic solvent. They are often separated by In this case, there is a problem that the thermally unstable volatile active ingredient changes in quality and also evaporates, resulting in a decrease in yield. As described above, when volatile active ingredients are extracted using an organic solvent, the quality of the volatile active ingredients and the yield of the volatile active ingredients are lowered.

[0005] Among volatile active ingredients, the aromatic components in liquid cultured cells in particular consist of a wide variety of volatile and water-soluble components with different polarities and boiling points, and a mixture of these components gives off a unique aroma. Further, the content of each component is small and coexists with a large amount of water. Since trace amounts of volatile components are easily dissipated at high temperatures, and water-soluble components are dissolved in a large amount of coexisting water, extraction methods using organic solvents are inefficient. Therefore, it is difficult to uniformly extract and separate such aroma components by the above-mentioned steam distillation method or extraction method using an organic solvent without impairing the characteristics of the aroma.

[0006] Furthermore, when searching for trace amounts of volatile active ingredients in a small amount of raw materials, it is important to extract the ingredients without altering their quality and to collect the extract without escaping it. However, the above-mentioned conventional extraction methods cause deterioration and a decrease in yield during the extraction process, so there is a problem that they are not suitable for use as an extraction method for searching for the true ingredients contained in raw materials.

[0007]

[Problems to be Solved by the Invention] In order to solve the above-mentioned problems, it is an object of the present invention to efficiently extract and separate volatile active ingredients from a material to be extracted in a high yield without altering their quality. Therefore, it is possible to extract and separate even trace amounts of volatile active ingredients, and even if the volatile active ingredients are aroma components in trace amounts, they can be extracted and separated without impairing the characteristics of the aroma. The purpose of this study is to propose a method for extracting volatile active ingredients.

[0008]

[Means for Solving the Problems] The present invention provides the following method for extracting volatile active ingredients.

(1) When extracting volatile active ingredients from a material to be extracted, supercritical carbon dioxide or carbon dioxide in a near supercritical state is used as an extractant at a temperature of 20 to 80°C and a pressure of 70 to 400 kgf/cm2 gauge. A method for extracting volatile active ingredients, characterized by extracting them using

(2) The method according to (1) above, characterized in that the extraction pressure during extraction of the volatile active ingredient is adjusted using a capillary.

(3) The method described in (1) or (2) above, wherein the volatile active ingredient is a fragrance ingredient.

(4) The method according to any one of (1) to (3) above, wherein the material to be extracted is a plant tissue or cell.

[0013] In the extraction method of the present invention, carbon dioxide used as an extractant for volatile active ingredients has a critical temperature (31.
1°C) and the critical pressure (75.3 kgf/cm2), or carbon dioxide in a state close to these critical points. Specifically, temperature 2
0 to 80°C, preferably 31.1 (critical temperature of carbon dioxide) to 70°C, more preferably 35 to 60°C, pressure 7
0-400kgf/cm2 gauge, preferably 75.3
(Critical pressure of carbon dioxide) ~300 kgf/cm2 gauge supercritical carbon dioxide or carbon dioxide in the vicinity of supercritical state, temperature and pressure are appropriately selected depending on the type of volatile active ingredient to be extracted.

[0014] As the extractant, it is preferable to use supercritical carbon dioxide whose temperature and pressure are both above the critical point. Carbon dioxide near supercritical carbon dioxide, where one or both of temperature and pressure are below the critical point, can also be used, but the extraction power will be smaller than that of supercritical carbon dioxide. On the other hand, carbon dioxide in a gaseous state outside the range of the supercritical carbon dioxide or carbon dioxide in the vicinity of the supercritical state has a low density and therefore has a low extraction power and cannot be used. Moreover, liquid carbon dioxide outside the range of the supercritical carbon dioxide or carbon dioxide in the vicinity of the supercritical state has low permeability to the material to be extracted such as plant cells, and is therefore undesirable because the extraction power is low.

The volatile active ingredients to be extracted in the extraction method of the present invention are not particularly limited, but include, for example, plant tissues or cells such as plant flowers, buds, fruits, seeds, branches, leaves, bark, trunks, rhizomes, etc.; Volatile substances that exist in extracted materials such as cultured cells and are used in the fields of pharmaceuticals, foods, fragrances, etc. can be mentioned. Preferred volatile active ingredients to be extracted include aroma components contained in the plant tissue or cells, cultured cells, etc., especially aroma components contained in trace amounts.

[0016] Moisture may be present in the extracted material used for extraction, and especially when extracting aroma components from cultured cells, the extracted material should be subjected to extraction without removing the water content by drying or other operations. Can be done.

In the extraction method of the present invention, an entrainer (a third component as an extraction aid) may be used in combination with carbon dioxide, if necessary. Preferred entrainers include methanol, ethanol, water (unnecessary if the raw material contains water), acetone, and the like. The content of these solvents as entrainers in the mixture of carbon dioxide and entrainer is usually 10 mol % or less, preferably 0.5 to 10 mol %. When using entrainer, extraction may be performed with carbon dioxide that does not contain entrainer and then further extraction with carbon dioxide that contains entrainer, or extraction may be performed with only carbon dioxide that contains entrainer.

[0018] The extraction method of the present invention using supercritical carbon dioxide or carbon dioxide in a near supercritical state is superior to extraction methods using organic solvents or steam distillation methods, which are conventionally frequently used for extracting plant-containing components. , has the following advantages:

[0019] There is no need to apply thermal energy during extraction. (2) There is no need to apply thermal energy when removing the extraction solvent after extraction, and the extraction components and extraction solvent can be easily separated. - Carbon dioxide itself has less chemical reactivity than alcohols, ketones, esters, and halogenated hydrocarbons that are commonly used in solvent extraction methods, so it is difficult to alter the extracted components. ■Since carbon dioxide itself is not toxic, there is no need to consider the toxicity of residual solvents when using extracted components as medicines or foods.

[0020]

[Example] Next, an example of the extraction method of the present invention will be explained with reference to the drawings.

FIG. 1 is a system diagram showing an example of the extraction method of the present invention. In the figure, 1 is an extraction tank that can be adjusted to a predetermined pressure and temperature, 2 is a separation tank, and both are connected to each other by a conduit 3 to form a circulation system. The extraction tank 1 is filled with a material to be extracted containing volatile active ingredients, and the volatile active ingredients are extracted by bringing carbon dioxide and an entrainer into contact with the extracted material. Here, carbon dioxide as an extractant is continuously supplied from a carbon dioxide supply source 4 and the entrainer from an entrainer storage tank 5 via a conduit 3 to the bottom of the extraction tank 1. At this time, carbon dioxide is pressurized from the carbon dioxide supply conduit 3a by a pressure booster pump 6a, heated by a heat exchanger 7a, and brought into a supercritical state at the above temperature and pressure or in a state near it, and is supplied to the extraction tank 1. . The entrainer is continuously supplied from the entrainer supply conduit 3b via a boost pump 6b, a stop valve 8a and a heat exchanger 7a. The entrainer supply conduit 3b is connected to the conduit 3 between the boost pump 6a and the heat exchanger 7a. The pressure in the extraction tank 1 is controlled by a capillary 9 and a stop valve 8b. Although the pressure may be adjusted using a pressure adjustment valve instead of the capillary 9, the dead volume and This is preferable because the pulsation of the extraction pressure can be reduced, thereby improving the extraction efficiency. In particular, it is preferable to use the capillary 9 when extracting a trace amount of a volatile active ingredient from a small amount of material to be extracted or when searching for a contained ingredient. The capillary 9 has an inner diameter of 50 to 300 μm, preferably 5 μm.
0 to 200 μm, length 50 to 500 mm, preferably 5
Those with a diameter of 0 to 300 mm can be used, and are appropriately selected depending on the extraction pressure.

The carbon dioxide containing volatile active ingredients extracted in the extraction tank 1 is continuously extracted from the upper part of the extraction tank 1 and introduced into the separation tank 2 via the heat exchanger 7b and the stop valve 8b. In the separation tank 2, carbon dioxide is made into a gaseous state at a pressure lower than that in the extraction tank 1, and the volatile active ingredients extracted by cooling with dry ice, acetone, etc. are made into a liquid or solid state. Separate the components and entrainer and carbon dioxide. The separated volatile active ingredient and entrainer are separated by stop valve 8c.
Collect through. On the other hand, the separated gaseous carbon dioxide is passed through a pressure-resistant check valve 10 through a conduit 3c for carbon dioxide recovery.
, is pressurized by a boost pump 6a, and then transferred to a heat exchanger 7.
a to bring it to a supercritical state or a state close to it, and supply it to the extraction tank 1 through the conduit 3, where it is used again as an extractant.

In the above extraction method, the material to be extracted containing volatile active ingredients may be continuously supplied to the extraction tank 1. In addition, instead of the separation tank 2, an extract collection container whose outside is cooled with dry ice, acetone, etc. is connected to the outlet of the stop valve 8b to prevent the extracted components from escaping along with carbon dioxide. It can also be used by connecting to 3. In this case, volatile active ingredients and entrainer are collected in the extract collection container, and carbon dioxide is released into the atmosphere from the outlet side of the extract collection container.

Next, Fig. 1 shows a method for extracting volatile aroma components from the callus of Picea acanthus using the callus of Picea acanthus as the material to be extracted containing volatile active ingredients.
The explanation will be based on.

[0025] The extraction tank 1 is filled with callus of Picea acanthus in a water-containing state or in a state in which water has been removed, preferably by crushing it. This extraction tank 1 can be adjusted to a predetermined pressure and temperature, and a conduit 3 is connected to the bottom of the extraction tank 1, from which supercritical carbon dioxide or supercritical carbon dioxide containing methanol, ethanol, etc. as an entrainer is pumped. Continuously supplied via pump 6a and heat exchanger 7a. The entrainer is continuously supplied from the entrainer storage tank 5 via the entrainer supply conduit 3b, the boost pump 6a and the stop valve 8a. In this extraction tank 1, the material to be extracted containing the components to be extracted, supercritical carbon dioxide, carbon dioxide in the vicinity of supercritical state, or these carbon dioxides containing entrainer (hereinafter sometimes simply referred to as extractant) are extracted. ) to extract volatile aroma components into the extractant. In this case, since the content of volatile aroma components in the callus of Picea abies is very small,
Preferably, a capillary 9 is used to regulate the pressure.

[0026] Next, the extractant in which the volatile aroma components have been dissolved is continuously extracted from the upper part of the extraction tank 1, lowered in pressure by the capillary 9, and then introduced into the separation tank 2 via the heat exchanger 7b and stop valve 8b. do. In the separation tank 2, the extract and carbon dioxide are separated by lowering the pressure while cooling with dry ice, acetone, etc. When a separated volatile aroma component or entrainer is used, the entrainer and volatile aroma component are recovered via the stop valve 8c. On the other hand, the separated carbon dioxide is transferred to the pressure-resistant check valve 1
0, the booster pump 6a and the heat exchanger 7a
After pressurizing and heating the fluid to make it a supercritical fluid again, it is circulated through the conduit 3 to the extraction tank 1 and reused. If some of the circulating carbon dioxide is lost, carbon dioxide is replenished from the carbon dioxide supply source 4. The extract collection container described above may be used instead of the separation tank 2.

[0027] By extracting with supercritical carbon dioxide as described above, the aroma components emitted by the callus of Picea spruce can be extracted and separated with high extraction efficiency without impairing the characteristics of the aroma.

Example 1 Picea spruce (Japanese spruce) emitting a fruit-like aroma that was kept frozen
Weighed 2.3 g of callus (Picea glehnii) with water in it (0.34 g after drying and removing the water content after extraction), and extracted volatile active ingredients using the extraction method shown in Figure 1. Extraction was performed. As the capillary 9, a capillary with an inner diameter of 200 μm and a length of 150 mm was used.

That is, callus of Picea acanthus was filled into an extraction tank 1 with an internal volume of 15 ml, supercritical carbon dioxide was used as an extractant, and the extraction pressure was 180 kgf/cm2 gauge.
Extraction was performed at an extraction temperature of 40° C. for 1 hour, and the extract was collected. Carbon dioxide was supplied continuously at a flow rate of 20 Nl/h. In the extraction operation, the weight of the charged material to be extracted was determined by weighing it using a balance, and the amount of carbon dioxide used was measured using a gas meter. For collection of the extract, an extract collection container whose outside was cooled with dry ice/acetone was used in place of the separation tank 2 to prevent the extracted aroma components from escaping along with carbon dioxide. This extract collection container is connected to a conduit 3 connected to the outlet of the stop valve 8b;
The extract was collected in the collection container. This collected extract is referred to as extract A.

[0030] Weighed 24.7 g of the same Picea callus as above while it contained moisture (3.7 g after the moisture content was removed by drying after extraction), and heated it to supercritical temperature in the same manner as above. An extraction test was conducted and the extract was collected. This collected extract is referred to as extract B.

[0031] In all of the above cases, extracts A and B collected in the collection container had a fruit-like aroma that was exactly the same as the fruit-like aroma emitted by the callus of Picea spruce, which was the extract to be extracted. Further, during extraction, the above-mentioned aroma was not observed at the outlet of the collection container, and the above-mentioned aroma was not observed at all from the callus of Picea abies taken out from the extraction tank 1 after extraction. These results indicate that the fruit-like aroma emitted by the callus of Picea abies could be completely extracted and separated by extraction using supercritical carbon dioxide.

Extract A was mainly used to analyze lipophilic components. That is, 500 μl of ether and 500 μl of water were injected into the collection container in which extract A had been collected using a syringe, and the collection container was rotated to dissolve the extract. The ether layer of the solution in the collection container was collected into a sample bottle. The aqueous layer was transferred to a stoppered test tube. Collection container 500μ
1 of water and 1000 μl of ether, and the washings were transferred to a stoppered test tube. This stoppered test tube was stoppered and shaken well to separate the ether layer. After adding 500 μl of ether again to the aqueous layer and shaking well, the ether layer was separated. All the ether layers separated by the above method were collected, dried over anhydrous sodium sulfate, and then subjected to gas chromatography.
It was analyzed using a mass spectrometer.

Extract B was used to analyze components with low boiling points and relatively hydrophilic properties. That is, the aqueous solution extracted with supercritical carbon dioxide in the collection container was directly analyzed using a gas chromatography/mass spectrometer.

[0034] The above two types of gas chromatography
1 to 3 μl of the sample to be subjected to mass spectrometry was injected into a gas chromatography/mass spectrometer. The conditions of the gas chromatography/mass spectrometer at that time were as follows. M-80B mass spectrometer manufactured by Hitachi, Ltd. Ionization energy: 70 eV Gas chromatography column: Fused silica capillary column UNBON HR
-101,50m x 0.25mmφ (manufactured by Shinwa Chemical Industry Co., Ltd.) Temperature conditions: 60℃ (5 minutes) 60-220℃ (2℃/min)

Ion chromatograms of the ether solution treated with extract A and the aqueous solution coexisting with extract B are shown in FIGS. 2 and 3.

As a result of structural analysis of peaks A to K components shown in FIG. 2, it was confirmed that they were the following substances. Peak A: Isoamyl alcohol peak B: Fatty acid ethyl ester peak C: Butyric acid peak D: Heptyl butyrate peak E: Fatty acid ethyl ester peak F: Fatty acid propyl ester peak G: Fatty acid ethyl ester peak H: Fatty acid ethyl ester peak I: Fatty acid methyl Ester peak J: Fatty acid ethyl ester peak K: Fatty acid ethyl ester

Further, as a result of structural analysis of peaks A to H components shown in FIG. 3, it was confirmed that they were the following substances. Peak A: Butenal peak B: Acetic acid peak C: Acetoin peak D: Ethylpropionic acid peak E: Fatty acid isobutyl ester peak F: Benzaldehyde peak G: Fatty acid isopropyl ester peak H: 1-
formylnaphthalene

[0038] From the above, it was confirmed that the main components of the fruit-like aroma emitted by the callus of Picea acanthus were the components shown in FIGS. 2 and 3.

Comparative Example 1 5.2 g (5.2 g) of callus of Picea sylvestris that produces the same aroma as the callus used in Example 1 was prepared in a moist state.
After extraction, 0.80g after drying and removing the moisture content
) The extract was weighed and filled into extraction tank 1, and extracted in the same manner as in Example 1, except that carbon dioxide was used as the extractant, the extraction pressure was 30 kgf/cm2 gauge, and the extraction temperature was 30°C. was collected in an extract collection container. The extract collected in this collection container did not have a fruit-like aroma similar to the fruit-like aroma emitted by the callus of Picea sylvestris, which is the material to be extracted.

Comparative Example 2 100 ml of distilled water was added to 13.4 g of callus of Picea spruce producing the same aroma as the callus used in Example 1, and the mixture was extracted by continuous distillation for 90 minutes with 30 ml of ether. The ether extract was dried over anhydrous sodium sulfate overnight, concentrated under reduced pressure, and then concentrated using a hand warm method, followed by gas chromatography/mass spectrometry. The conditions of the gas chromatography/mass spectrometer at that time were as follows. M-80B mass spectrometer manufactured by Hitachi, Ltd. Ionization energy: 70 eV Gas chromatography column: Silicone OV-101, 50 m x 0.25 mm
φ (manufactured by Shinwa Chemical Industry Co., Ltd.) Temperature conditions: 60°C (5 minutes) 60-220°C (2°C/min)

As a result, pyridine, benzaldehyde,
Aromatic compounds such as ethyl benzoate, vanillin, naphthalene ethyl acetate, ethoxybenzaldehyde, acetic acid, acyl alcohol, various aldehydes with 6 to 10 carbon atoms, alcohol, fatty acids with 12 to 18 carbon atoms and their methyl or ethyl esters, acyl Acetate, carbon number 1
Although 3 to 16 linear hydrocarbons were identified, the extract showed no aroma and failed to capture fruity aromas.

Comparative Example 3 After thawing 102.8 g of liquid cultured cells of Picea acanthus that produce the same aroma as the callus used in Example 1, they were soaked in 200 m of water.
1 was added to make the mixture uniformly suspended. Divide this into 4 parts, 7
5 ml portions were dispensed into 500 ml round bottom flasks, rapidly frozen with acetone/dry ice, and then freeze-dried. 295 ml of recovered water (required time: 6 hours) was obtained and extracted with dichloromethane. In addition, the obtained freeze-dried residue is
Furthermore, the water recovered after the dichloromethane extraction was added, and volatile components were collected by freeze-drying in the same manner as above. The two batches were combined and concentrated using a Guderner-Danish concentrator (manufactured by Shibata Kagaku Kikai Kogyo Co., Ltd.) to obtain 3 mg of a concentrate.

[0043] The fruit-like aroma as observed in Example 1 was not observed in this concentrate. Moreover, this concentrate was used for gas chromatography/mass spectrometry. The conditions of the gas chromatography/mass spectrometer are as follows. M-80B mass spectrometer manufactured by Hitachi, Ltd. Ionization energy: 70 eV Gas chromatography column: PEG 20M bonded (25 m x 0.25 mm
ID, ) (manufactured by Shinwa Chemical Industry Co., Ltd.) Temperature conditions: 45°C (4 minutes) 45-230°C (3°C/min)

According to the results of gas chromatography/mass spectrometry, acetoin, acetoin alcohol, 1-octen-3-ol, butene-1,4-olide, menthol, methyl carbinol, isopentenol, 4-
Low-boiling substances such as methyl-butyric acid were detected, but no components thought to be related to the fruity aroma could be identified.

[0045] According to the above-mentioned Example 1 and Comparative Examples 1 to 3, the extraction method using supercritical carbon dioxide in Example 1 does not impair the quality of volatile aroma components compared to the freeze-drying method and the steam distillation method. It can be seen that extraction and separation can be performed efficiently without any problems.

[0046] The volatile aroma components of the callus of Picea acanthus are composed of a plurality of components and are responsible for the characteristic scent, and some of these components are contained in trace amounts. In Comparative Example 2, there was concern that the aroma components would deteriorate at high temperatures;
When separating the oil and water layers of the distillate, the water-soluble components of the target aromatic components are further extracted from the water layer.
Separation operation is required. In Comparative Example 3, the contained components are frozen together with the contained water, and the contained components are collected by sublimation or evaporation as the water sublimates under reduced pressure.For components with high boiling points, the dissolving power of carbon dioxide The extraction power is smaller than that of the extraction method of the present invention, which extracts the contained components. On the other hand, according to the extraction method of the present invention, the target aroma components can be efficiently extracted and separated using the appropriate extraction power of supercritical carbon dioxide, and the target aroma components can be extracted and separated without any post-treatment of the extract. Components can be extracted and separated.

[0047]

[Effects of the Invention] As described above, according to the present invention, volatile active ingredients are extracted using supercritical carbon dioxide or carbon dioxide in the vicinity of supercritical state, so that volatile active ingredients cannot be altered. It can be extracted and separated with high yield and efficiency. Therefore, even trace amounts of volatile active ingredients can be extracted and separated, and even aromatic components can be extracted and separated without impairing their aroma characteristics.

[Brief explanation of the drawing]

FIG. 1 is a system diagram showing an example of the extraction method of the present invention.

FIG. 2 is a graph showing the results of Example 1.

FIG. 3 is a graph showing the results of Example 1.

[Explanation of symbols]

1 Extraction tank 2 Separation tank 3, 3a, 3b, 3c conduit 4. Carbon dioxide supply source 5 Entrainer storage tank 6a, 6b Boost pump 7a, 7b Heat exchanger 8a, 8b, 8c Stop valve 9 Capillary 10 Pressure resistant check valve

Claims (4)

[Claims] Claim 1: When extracting volatile active ingredients from a material to be extracted, an extractant is used at a temperature of 20-80°C and a pressure of 70-80°C.
1. A method for extracting volatile active ingredients, characterized by extraction using supercritical carbon dioxide or carbon dioxide in a near supercritical state with a gauge of 400 kgf/cm2. 2. The method according to claim 1, wherein the extraction pressure during extraction of the volatile active ingredient is adjusted using a capillary. 3. The method according to claim 1 or 2, wherein the volatile active ingredient is a fragrance ingredient. 4. The method according to claim 1, wherein the extract is a plant tissue or cell.