JP2019037913A - Extraction method of terpenes - Google Patents

Abstract

To provide an extraction method of terpenes capable of extracting efficiently terpenes from a vegetable.SOLUTION: An extraction method of terpenes includes an extraction step ST11 for mixing an extraction solvent containing ionic liquid capable of dissolving cellulose with a vegetable containing terpenes, and extracting terpenes from the vegetable together with the extraction solvent as extract, and a recovery step ST12 for mixing the extract with an organic solvent capable of dissolving terpenes, then, separating the extract from the organic solvent by centrifugal separation, and recovering the terpenes from the organic solvent.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a method for extracting terpenes, for example, a method for extracting terpenes in which terpenes contained in a plant are extracted with an ionic liquid.

  Synaropicrin is a natural organic compound of sesquiterpene lactone isolated and determined from artichoke in 1960. Sinalopicrin has an action of suppressing transcriptional activity of NF-κb (nuclear factor κb). Sinalopicrin prevents “photoaging” caused by stimulating the transcriptional activity of NF-κb by external stimuli such as ultraviolet rays and inflammatory cytokines, suppresses an increase in melanin pigment, and can be expected to have a whitening effect. In addition, quinicin (Cnicin) is a natural organic compound of sesquiterpene lactone, which was isolated and determined in 1959 from Santorium. Sinalopicrin and quinicin have pharmacological activity against a parasitic protozoan called Trypanosoma brucei that causes African sleeping sickness, and are also expected as therapeutic drugs for African sleeping sickness.

  By the way, in recent years, research on an ionic liquid (also referred to as “ionic liquid”), which is an organic salt having a low melting point composed only of ions, has attracted attention. The ionic liquid has high polarity as a result of being composed only of ions, and has characteristics such as non-volatility, high thermal stability, and chemical stability. Some ionic liquids are also referred to as “room temperature molten salts” or “room temperature molten salts” that are in a liquid state near room temperature. In addition, some ionic liquids can dissolve cellulose that is difficult to dissolve in common organic solvents and water, and a technique for extracting caffeoylquinic acid contained in coffee beans using ionic liquids has been proposed. (For example, see Patent Document 1 and Non-Patent Document 1).

JP2015-209401A

Toyonobu Usuki, Shingo Ondda, Masahiro Yoshizawa-Fujita, Masahiro Rikukawa "Use of [C4mim] Cl for Efficient Extraction of Caffeoylquinic Acids from Sweet Potato Leaves" Scientific Reports 2017, 7, 6890.

  Many terpenes such as sinalopicrin and quinicin have not been fully synthesized. Development of an extraction method capable of efficiently extracting terpenes from plants is desired.

  This invention is made | formed in view of such a situation, and it aims at providing the extraction method of terpenes which can extract terpenes from a plant efficiently.

  The method for extracting terpenes according to the present invention comprises an extraction solvent containing an ionic liquid capable of dissolving cellulose and a plant containing terpenes, and extracting the terpenes from the plant together with the extraction solvent as an extract. And a recovery step of mixing the extract and the organic solvent capable of dissolving the terpenes, separating the extract and the organic solvent by centrifugation, and recovering the terpenes in the organic solvent; including.

  In the extraction method of terpenes according to the present invention, it is preferable that the extraction liquid, the organic solvent, and water are mixed in the recovery step.

  In the extraction method of terpenes according to the present invention, the extraction solvent is a mixed solvent of the ionic liquid and a polar solvent containing at least one selected from the group consisting of acetones, alcohols and esters. Is preferred.

  In the extraction method of terpenes according to the present invention, in the extraction step, the temperature during the extraction is preferably 15 ° C. or more and 60 ° C. or less.

  In the terpene extraction method according to the present invention, the mixed solvent preferably has a mass ratio of the ionic liquid to the polar solvent (ionic liquid: polar solvent) in the range of 5: 1 to 1: 5. .

  In the method for extracting terpenes according to the present invention, the organic solvent is further added to and mixed with the extract separated in the recovery step, and then the extract and the organic solvent are separated by centrifugation. It is preferable to include a regeneration step for regenerating the ionic liquid.

（式（１）中、Ｒは、炭素数１以上５以下のアルキル基を表す。） In the terpene extraction method according to the present invention, the ionic liquid is preferably represented by the following general formula (1).

(In formula (1), R represents an alkyl group having 1 to 5 carbon atoms.)

  In the method for extracting terpenes according to the present invention, the terpenes are preferably sesquiterpenes.

  ADVANTAGE OF THE INVENTION According to this invention, the extraction method of terpenes which can extract terpenes from a plant efficiently is realizable.

FIG. 1 is a flowchart showing an outline of a terpene extraction method according to an embodiment of the present invention. FIG. 2 is a diagram showing the extraction rate of the extraction method of terpenes according to Examples and Comparative Examples of the present invention. FIG. 3 is a diagram showing the relationship between the extraction time and the extraction rate of the extraction method of terpenes according to Examples and Comparative Examples of the present invention. FIG. 4 is a diagram showing the relationship between the number of reuses of the ionic liquid and the extraction rate in the extraction method of terpenes according to Examples and Comparative Examples of the present invention. FIG. 5 is a diagram showing the extraction rate of the extraction method of terpenes according to Examples and Comparative Examples of the present invention.

  Hereinafter, an embodiment of the present invention will be described in detail with reference to the accompanying drawings. In addition, this invention is not limited at all by the following embodiment.

  FIG. 1 is a flowchart showing an outline of a terpene extraction method according to the present embodiment. As shown in FIG. 1, the extraction method of terpenes according to the present embodiment mixes an extraction solvent containing an ionic liquid capable of dissolving cellulose and a plant containing terpenes, and terpenes from the plant together with the extraction solvent. Extraction step ST11 for extraction as an extract and an extraction solvent and an organic solvent capable of dissolving terpenes are mixed, and then the extract and the organic solvent are separated by centrifugation to recover the terpenes in the organic solvent. Including ST12. Hereinafter, a method for extracting terpenes according to the present embodiment will be described in detail.

  In the extraction step ST11, an extraction solvent containing an ionic liquid capable of dissolving cellulose and a plant containing terpenes are mixed for a predetermined time. Moreover, in extraction process ST11, the terpenes contained in the plant mixed for a predetermined time with the extraction solvent are extracted as an extract together with the extraction solvent.

  The extraction solvent contains an ionic liquid capable of dissolving cellulose (hereinafter, also simply referred to as “ionic liquid”). The ionic liquid is not limited as long as it can dissolve cellulose. An ionic liquid refers to a liquid which is fluid over a wide temperature range and is completely composed of ions. As an ionic liquid, the salt which will be in a liquid state at 100 degrees C or less (for example, below room temperature) is shown, for example.

  The ionic liquid is a salt and has a cation and an anion. Examples of the cation of the ionic liquid used as the extraction solvent include organic cations such as a quaternary ammonium cation, an imidazolium cation, and a pyridinium cation. These organic cations may be used alone or in combination of two or more.

  Examples of the cation of the ionic liquid include 1-butyl-3-methylimidazolium cation, 1-ethyl-3-methylimidazolium cation, and 1-allyl-3-methylimidazolium cation. Among these, 1-butyl-3-methylimidazolium cation and 1-ethyl-3-methylimidazolium cation are preferable from the viewpoint of efficiently extracting terpenes from plants, and 1-ethyl-3-methylimidazolium cation is preferable. Is more preferable.

Examples of the anion of the ionic liquid include BF ₄ ⁻ , PF ₆ ⁻ , AsF ₆ ⁻ , SbF ₆ ⁻ , AlCl ₄ ⁻ , HSO ₄ ⁻ , ClO ₄ ⁻ , CH ₃ SO ₃ ⁻ , and CH ₃ SO ₄ ^−. , CH ₃ C ₆ H ₄ SO ₃ ⁻ , CF ₃ SO ₃ ⁻ , CF ₃ SO ₄ ⁻ , CF ₃ CO ₂ ⁻ , C ₂ F ₅ CO ₂ ⁻ , (CF ₃ SO ₂ ) ₂ N ⁻ , (C _{2 ^{F 5 SO 2) 2 N -}} , phosphonic ^{_{acid, (CH 3 O) 2 PO}} 2 -, (CH 3 O) CH 3 PO 2 -, (CH 3 O) C 2 H 5 OPO 2 -, (CH 3 O ) HPO ₂ ⁻ , (C ₂ H ₅ O) ₂ PO ₂ ⁻ , (C ₂ H ₅ O) CH ₃ PO ₂ ⁻ , (C ₂ H ₅ O) C ₂ H ₅ PO ₂ ⁻ , (C ₂ H ₅ O) HPO ₂ ^- phosphate ions, such as, ^Cl -, Br ^{-, -} a halide ion, the number 1 to 3 carboxylic acid ion carbons such as formate ion (HCOO ^-), perchlorate ^ion, CN ^-, SCN ^-, OCN -, ^ONC -, _{N 3} ^- pseudohalide such as Examples thereof include pseudohalide ions such as ions, cyanamide ions (NCN ⁻ ), and dicyanamide ions (N (CN) ₂ ⁻ ).

Among these, as the anion of the ionic liquid, a phosphate anion is preferable, and an alkylphosphonate anion ((RO) HPO ₂ ⁻ ) (R has 1 to 5 branches) having 1 to 5 carbon atoms. Represents an optionally substituted alkyl group). Examples of the alkyl phosphonate anion having 1 to 5 carbon atoms include methyl phosphonate anion ((CH ₃ O) HPO ₂ ⁻ ), ethyl phosphonate anion ((C ₂ H ₅ O) HPO ₂ ⁻ ), and propyl phosphonate anion ((C ₂ H ₅ O) HPO ₂ ⁻ ) is more preferable, and methylphosphonate anion ((CH ₃ O) HPO ₂ ⁻ ) is particularly preferable.

  Examples of the combination of cation and anion of the ionic liquid include 1-ethyl-3-methylimidazolium chloride, 1-ethyl-3-methylimidazolium methylphosphonate, 1-ethyl-3-methylimidazolium ethylphosphonate. 1-ethyl-3-methylimidazolium propylphosphonate, 1-ethyl-3-methylimidazolium butylphosphonate, 1-ethyl-3-methylimidazolium pentylphosphonate, 1-ethyl-3-methylimidazolium dicyanamide and the like Can be mentioned.

（式（１）中、Ｒは、炭素数１以上５以下のアルキル基を表す。） As the ionic liquid, from the viewpoint of efficiently extracting terpenes from plants, among the combinations of the cation and the anion described above, those represented by the following general formula (1) are preferable. In the following general formula (1), R is preferably an alkyl group having 1 to 5 carbon atoms, and more preferably an alkyl group having 1 to 3 carbon atoms. Examples of R include a methyl group, an ethyl group, a propyl group, an isopropyl group, an n-butyl group, an isobutyl group, an s-butyl group, a t-butyl group, and a pentyl group. These alkyl groups having 1 to 5 carbon atoms may be unsubstituted or may have a substituent such as a hydroxyl group and a halogen atom. Among these, as R, a methyl group, an ethyl group, a propyl group and an isopropyl group are preferable, a methyl group and an ethyl group are more preferable, and a methyl group is still more preferable.

(In formula (1), R represents an alkyl group having 1 to 5 carbon atoms.)

As the ionic liquid, 1-ethyl-3-methylimidazolium methylphosphonate, 1-ethyl-3-methylimidazolium ethylphosphonate and 1-ethyl-3-methylimidazolium propylphosphonate are more preferable, and 1-ethyl-3- More preferred is methylimidazolium methylphosphonate ([C ₂ mim] [(MeO) (H) PO ₂ ]). In addition, what can be obtained commercially may be used for the ionic liquid as an extraction solvent, and what was synthesize | combined by the well-known method may be used.

  Moreover, as an extraction solvent, it is preferable to use the mixed solvent containing an ionic liquid from a viewpoint of improving the extraction rate of terpenes more. For example, examples of the mixed solvent containing an ionic liquid include a mixed solvent containing a supercritical fluid such as water, an organic solvent, and supercritical carbon dioxide in addition to the ionic liquid. Here, as the organic solvent, for example, polar solvents such as ketones, alcohols, esters, ethers and nitriles are preferable.

  As the alcohol, an alcohol compound having 1 to 5 carbon atoms is preferable, and an alcohol compound having 1 to 3 carbon atoms is more preferable. Examples of alcohols include methanol, ethanol, propyl alcohol, isopropanol, butanol, isobutanol, sec-butanol, t-butanol, and pentanol. Among these, as alcohols, methanol, ethanol, propanol and isopropanol are preferable, and ethanol is more preferable.

  As the ketones, ketone compounds having 1 to 5 carbon atoms are preferable. Examples of ketones include acetone, 2-butanone, 2-pentanone, and 3-pentanone. Among these, acetone and 2-butanone are preferable, and acetone is more preferable.

  As the esters, ester compounds having an ester group having 1 to 5 carbon atoms are preferable, and ester compounds having an ester group having 1 to 3 carbon atoms are more preferable. Examples of the esters include methyl acetate, ethyl acetate, propyl acetate, butyl acetate, pentyl acetate, methyl propionate, ethyl propionate, propyl propionate, butyl propionate and pentyl propionate. Among these, as esters, methyl acetate, ethyl acetate, propyl acetate, methyl propionate, ethyl propionate and propyl propionate are preferable, ethyl acetate and ethyl propionate are more preferable, and ethyl acetate is still more preferable.

  As the ethers, ether compounds having 4 to 10 carbon atoms are preferable. Examples of ethers include diethyl ether, methyl propyl ether, ethyl propyl ether, dipropyl ether, methyl butyl ether, ethyl butyl ether, propyl butyl ether, dibutyl ether, tetrahydrofuran, 1,3-dioxane and 1,4-dioxane 2-butanone. , 2-pentanone, 3-pentanone and the like.

  As the nitriles, nitrile compounds having 2 to 5 carbon atoms are preferable. Examples of nitriles include acetonitrile, propionitrile, butyronitrile, and the like.

  Moreover, the mixing ratio of the ionic liquid and the polar solvent when using the mixed solvent as the extraction solvent is such that the mass ratio (ionic liquid: polar solvent) is 5: 1 to 1 from the viewpoint of further improving the extraction rate of terpenes. : 5, preferably 4: 1 to 1: 4, more preferably 3: 1 to 1: 3, and 1: 2 to 2: 1. A range is particularly preferred. If the mass ratio (ionic liquid: polar solvent) is in the range of 5: 1 to 1: 5, the extraction method of terpenes is an ionic liquid having a high viscosity without impairing the solubility of plant cellulose by the ionic liquid. Is diluted with a polar solvent to lower the viscosity of the mixed solvent, so that the extraction rate of terpenes can be further improved.

In this Embodiment, the terpene which exists in the inside of the cellulose which comprises the cell wall of a plant is extracted with an ionic liquid by using the ionic liquid which melt | dissolves a cellulose. Therefore, the terpenes are not particularly limited as long as they can be extracted with an ionic liquid, and various terpenes such as monoterpenes, sesquiterpenes, diterpenes, and terpenoids derived from these can be used. Among these, as a terpene, from the viewpoint that terpenes can be efficiently extracted from plants, sesquiterpenes and sesquiterpenes that are derivatives thereof are preferable, sesquiterpene lactone derivatives are more preferable, and synaropicrin represented by the following formula (2). And the quinicine shown to following formula (3) is still more preferable.

  The plant added to the extraction solvent is not particularly limited as long as it contains terpenes. Examples of the plant in the extraction step ST11 include sweet potato leaves, spring chrysanthemum, burdock, burdock, mugwort, scallops, artichokes, and scallops, and solanaceae such as eggplant and potato, coffee, morohaya, ensai, and pineapple. Examples include leaves. Among these, asteraceae plants such as burdock, burdock, mugwort, nigana, artichoke and santori are preferred, and artichoke and santori are more preferred from the viewpoint that terpenes can be efficiently extracted from the plant.

  Moreover, as a plant in extraction process ST11, a part in arbitrary plants containing terpenes may be used, and all the plants may be used. As a part of the plant, for example, it is preferable to use a plant leaf, and it is more preferable to use a dry, powdered one. Moreover, as a plant in extraction process ST11, it is preferable to use the ground plant from a viewpoint of improving extraction efficiency, and it is more preferable to use a powdery thing. Any method may be used as a method for pulverizing the plant. In addition, you may purchase and use what was pulverized originally.

  There is no restriction | limiting in particular as extraction temperature in extraction process ST11 if it is the range which can extract terpenes from a plant. The extraction temperature in the extraction step ST11 is preferably 15 ° C. or more and 60 ° C. or less, more preferably 20 ° C. or more and 55 ° C. or less, and more preferably 25 ° C. or more, from the viewpoint of improving the extraction rate of terpenes from plants. More preferably, it is 50 degrees C or less. In particular, in this embodiment, since terpenes can be efficiently extracted from plants even near room temperature (for example, around 25 ° C.), the cost required for extraction such as equipment and time accompanying heating of the extraction solvent is reduced. It becomes possible to do.

  The extraction time for mixing the extraction solvent and the plant in the extraction step ST11 is not particularly limited as long as the terpene can be extracted from the plant. The extraction time is, for example, 10 minutes or more and 200 minutes or less. The extraction time is preferably 15 minutes or more and 180 minutes or less, more preferably 30 minutes or more and 150 minutes or less, and more preferably 60 minutes or more and 140 minutes or less from the viewpoint of efficiently extracting terpenes from plants. Is more preferable, and 110 to 130 ° C. is particularly preferable. In particular, in the present embodiment, the terpene can be extracted efficiently only by performing the extraction for about 60 minutes near the room temperature described above, so that the cost such as time required for the extraction of the terpene from the plant can be reduced. In addition, the extraction can be performed most efficiently by setting the extraction time to about 120 minutes.

  The mass ratio of the plant and the extraction solvent in the extraction step ST11 is not particularly limited as long as the terpene can be extracted from the plant. The mass ratio between the plant and the extraction solvent (plant: extraction solvent) is in the range of 1:20 to 1: 3 from the viewpoint of efficiently extracting the terpenes by efficiently stirring the plant and the extraction solvent. Is preferably in the range of 1:15 to 1: 5, more preferably in the range of 1: 12.5 to 1: 7.5. In addition, the mass of the plant may be dried or may not be dried.

  In the recovery step ST12, the extract and an organic solvent capable of dissolving terpenes are mixed for a predetermined time. In the recovery step ST12, the mixed extract and the organic solvent are centrifuged for a predetermined time to recover the terpenes in the organic solvent, and then the extract and the organic layer are separated by decantation.

  As an organic solvent capable of dissolving terpenes (hereinafter, also simply referred to as “organic solvent”), a solvent that can be separated from an ionic liquid by centrifugation is used. As an organic solvent, the polar solvent used for the mixed solvent mentioned above is mentioned, for example. Among these, as the organic solvent, esters are preferable, and ethyl acetate is more preferable.

  Further, the mass of the organic solvent with respect to the terpenes in the recovery step ST12 is not limited as long as the terpenes can be extracted from the extract. The mass of the organic solvent relative to the terpenes is preferably 5 parts by mass or more and 25 parts by mass or less, preferably 10 parts by mass or more and 20 parts by mass with respect to 1 part by mass of the plant, from the viewpoint of efficiently recovering the terpenes from the extract. More preferably, it is 1 part by mass or more and 17.5 parts by mass or less.

  The mixing time of the extract and the organic solvent in the recovery step ST12 is not particularly limited as long as terpenes can be recovered from the extract. The mixing time is preferably from 1 minute to 20 minutes, more preferably from 2 minutes to 10 minutes, more preferably from 3 minutes to 7.5 minutes, from the viewpoint of efficiently recovering terpenes from the extract. More preferably, it is as follows.

  The mixing temperature of the extract and the organic solvent in the recovery step ST12 is not particularly limited as long as terpenes can be recovered from the extract. The mixing temperature in the recovery step ST12 can be performed at room temperature such as 10 ° C. or higher and 30 ° C. or lower.

  In the recovery step ST12, water may be added and mixed in addition to the extract and the organic solvent. This reduces the solubility of the organic solvent in the extract containing the ionic liquid and improves the separation between the extract containing the ionic liquid and the organic layer containing the terpenes and the organic solvent. It will be possible to recover the product even more efficiently. There is no restriction | limiting in particular as water, Various waters, such as a tap water, ion-exchange water, and distilled water, can be used.

  The mass of the water added in the recovery step ST12 is not limited as long as the separation between the extract and the organic layer can be improved. The mass of water is preferably 0.1 parts by mass or more and 10 parts by mass or less, based on 1 part by mass of the plant, from the viewpoint of improving the liquid separation between the extract and the organic layer. The amount is more preferably 8 parts by mass or less, and still more preferably 2 parts by mass or more and 6 parts by mass or less.

  In the collection step ST12, the extraction liquid containing the ionic liquid and the extracted plant is separated from the organic layer containing the terpenes and the organic solvent by centrifuging using a general centrifugal separator. The centrifugation time is not particularly limited as long as the extract can be separated from the organic layer. The time for centrifugation is preferably 10 seconds to 5 minutes, more preferably 30 seconds to 3 minutes. Further, the temperature for centrifugation is the same as the mixing temperature described above. As described above, in the present embodiment, the extraction liquid containing the ionic liquid and the plant after extraction can be efficiently separated from the organic layer in a short time and under mild temperature conditions by centrifugation. Not only can the equipment associated with the extraction be simplified and the time can be shortened, but also the industrial scale-up can be facilitated by improving the liquid separation property.

  The terpenes contained in the organic layer thus obtained are measured, for example, by analyzing them using reverse phase high performance liquid chromatography (HPLC: High Performance Liquid Chromatography) absolute calibration curve method. be able to.

  In the extraction method of terpenes according to the present embodiment, a regeneration step of regenerating the ionic liquid so as to be reusable may be performed after the recovery step ST12.

  In the regeneration step, an organic solvent similar to that used in the recovery step ST12 is added to the extract containing the ionic liquid separated in the recovery step ST12 and mixed for a predetermined time. The temperature and time during mixing are the same as in the recovery step ST12. In the regeneration step, the ionic liquid and the organic solvent are separated by decantation by centrifuging the mixed extract and the organic solvent for a predetermined time. The time for centrifugation is the same as in the recovery step ST12. In the regeneration step, after repeating these steps a predetermined number of times, the ionic liquid is regenerated by performing filtration and concentration of volatile components.

  The ionic liquid regenerated in this way can be reused at least nine times without reducing the extraction rate. Thereby, since the extraction method of terpenes can reduce the ionic liquid discharged | emitted after extraction of terpenes, it can reduce the cost accompanying extraction of terpenes. The terpene extraction method eliminates the need to discard the ionic liquid every time the terpenes are extracted, and can be regenerated simply by mixing and centrifuging the ionic liquid and the organic solvent. The wastewater discharged along with this can also be reduced. In addition, the extraction method of terpenes can be regenerated by simply mixing and centrifuging the ionic liquid and the organic solvent, so that the ionic liquid can be regenerated without preparing special equipment, and the liquid separation property is also improved. It is possible to cope with scale-up.

  As described above, according to the above-described embodiment, terpenes obtained by extracting terpenes from plants using an ionic liquid capable of dissolving cellulose are extracted by dissolving them in an organic solvent and then centpenes by centrifugation. The organic layer containing is separated and recovered. Thereby, since the extraction method of terpenes can collect terpenes from a plant with a high extraction rate, it becomes possible to extract terpenes efficiently. Moreover, the extraction method of terpenes which can extract terpenes efficiently can be implement | achieved compared with the case where other organic solvents and water are used as an extraction solvent. Furthermore, the separation between the extract containing the ionic liquid and the organic solvent containing the terpenes is improved by centrifugation, and a separation operation is not required, so that scale-up can be easily handled. Furthermore, by using an ionic liquid as an extraction solvent, a terpene extraction method that can be repeatedly used with the ionic liquid as an extraction solvent can be realized.

  Hereinafter, the present invention will be described in more detail on the basis of Examples and Comparative Examples performed to clarify the effects of the present invention. In addition, this invention is not limited at all by the following examples and comparative examples.

  First, the present inventors examined the relationship between the extraction solvent, the extraction temperature, and the extraction rate for extraction of synaropicrin using an ionic liquid. The contents examined by the inventors will be described below.

Example 1
Sample of artichoke leaf (0.05 g) pulverized by a mixer and 1-ethyl-3-methylimidazolium methylphosphonate (hereinafter simply referred to as “ionic liquid” in Examples and Comparative Examples) (0.5 g) In addition to the tube, it was stirred at 25 ° C. for 60 minutes. Thereafter, distilled water (0.214 g) and ethyl acetate (0.714 g) were added and stirred for 5 minutes, and then the extract was separated from the organic layer (ethyl acetate layer) by centrifuging for 1 minute. The layer was collected by decantation. The operations of adding ethyl acetate, centrifuging and separating the organic layer were repeated three times. The obtained organic layer was analyzed by reversed-phase liquid chromatography under the following analysis conditions, and the extraction rate of synaropicrin was quantified using an absolute calibration curve method. As a result, the extraction rate of synaropicrin was 0.53%. The results are shown in Table 1 and FIG.
(HPLC analysis conditions)
Column: YMC-Pack ODS-AM (150 × 4.6 mm)
Solvent: Measured by the gradient method under the following conditions.
0 min to 5 min; acetonitrile / distilled water = 2: 98
5 min to 30 min; acetonitrile / distilled water = 100: 0
30 min to 40 min; acetonitrile / distilled water = 100: 0
40 min to 45 min; acetonitrile / distilled water = 2: 98
45 min to 55 min; acetonitrile / distilled water = 2: 98
Flow rate: 1.0 mL / min
Column temperature: 40 ° C
UV detection: 228 nm

(Example 2)
The extraction rate of synaropicrin was quantified in the same manner as in Example 1 except that artichoke leaves (0.05 g) and ionic liquid (0.5 g) pulverized by a mixer were stirred at 50 ° C. for 60 minutes. As a result, the extraction rate of synaropicrin was 0.54%. The results are shown in Table 1 and FIG.

(Example 3)
In place of the ionic liquid (0.5 g), the extraction rate of synaropicrin was increased in the same manner as in Example 1 except that a mixed solvent obtained by mixing ionic liquid (0.25 g) and acetone (0.25 g) was used. Quantified. As a result, the extraction rate of synaropicrin was 0.71%. The results are shown in Table 1 and FIG.

Example 4
In place of the ionic liquid (0.5 g), the extraction rate of synaropicrin was increased in the same manner as in Example 2 except that a mixed solvent obtained by mixing ionic liquid (0.25 g) and acetone (0.25 g) was used. Quantified. As a result, the extraction rate of synaropicrin was 0.74%. The results are shown in Table 1 and FIG.

(Example 5)
Instead of the ionic liquid (0.5 g), the extraction rate of synaropicrin was increased in the same manner as in Example 2 except that a mixed solvent obtained by mixing ionic liquid (0.25 g) and ethanol (0.25 g) was used. Quantified. As a result, the extraction rate of synaropicrin was 0.59%. The results are shown in Table 1 and FIG.

(Example 6)
Extraction rate of synaropicrin in the same manner as in Example 2 except that a mixed solvent obtained by mixing ionic liquid (0.25 g) and ethyl acetate (0.25 g) was used instead of ionic liquid (0.5 g). Was quantified. As a result, the extraction rate of synaropicrin was 0.67%. The results are shown in Table 1 and FIG.

(Example 7)
In place of the ionic liquid (0.5 g), the extraction rate of synaropicrin was increased in the same manner as in Example 2 except that a mixed solvent obtained by mixing ionic liquid (0.375 g) and ethanol (0.125 g) was used. Quantified. As a result, the extraction rate of synaropicrin was 0.74%. The results are shown in Table 1 and FIG.

(Example 8)
In place of the ionic liquid (0.5 g), the extraction rate of synaropicrin was increased in the same manner as in Example 2 except that a mixed solvent obtained by mixing ionic liquid (0.125 g) and ethanol (0.375 g) was used. Quantified. As a result, the extraction rate of synaropicrin was 0.59%. The results are shown in Table 1 and FIG.

(Comparative Example 1)
Artichoke leaves (0.05 g) and acetone (0.5 g) pulverized by a mixer were added to a sample tube and stirred at 25 ° C. for 60 minutes. Thereafter, the mixture was further stirred for 15 minutes, and then centrifuged for 10 minutes to separate the organic layer (acetone) from the solid artichoke leaf, and the organic layer was recovered by decantation. The obtained organic layer was analyzed in the same manner as in Example 1 to quantify the extraction rate of synaropicrin. As a result, the extraction rate of synaropicrin was 0.46%. The results are shown in Table 1 and FIG.

(Comparative Example 2)
The extraction rate of synaropicrin was quantified in the same manner as in Comparative Example 1 except that artichoke leaves (0.05 g) and acetone (0.5 g) ground by a mixer were stirred at 50 ° C. for 60 minutes. As a result, the extraction rate of synaropicrin was 0.53%. The results are shown in Table 1 and FIG.

(Comparative Example 3)
As in Comparative Example 2, except that ethanol (0.5 g) was used instead of acetone (0.5 g). The extraction rate of synaropicrin was quantified. As a result, the extraction rate of synaropicrin was 0.35%. The results are shown in Table 1 and FIG.

(Comparative Example 4)
As in Comparative Example 2, except that ethyl acetate (0.5 g) was used instead of acetone (0.5 g). The extraction rate of synaropicrin was quantified. As a result, the extraction rate of synaropicrin was 0.43%. The results are shown in Table 1 and FIG.

  As can be seen from Table 1 and FIG. 2, when the ionic liquid is used alone, the extraction temperature is 25 ° C., the extraction rate is 0.53% (Example 1), and the extraction temperature is 50 ° C. Was 0.54% (Example 2), and a high extraction rate was obtained in all cases. In addition, when extraction was performed using a 1: 1 mixed solvent of an ionic liquid and a polar solvent (acetone, ethanol, ethyl acetate) (Examples 3 to 6), The extraction rate was further improved in comparison (Example 3: 0.71%, Example 4: 0.74%, Example 5: 0.59%, Example 6: 0.67%). In addition, a 3: 1 mixed solvent and a 1: 3 mixed solvent of an ionic liquid and an organic solvent (ethanol) are used (Examples 7 and 8) as compared with the case where only the ionic liquid is used. Thus, the extraction rate was further improved (Example 7: 0.74%, Example 8: 0.59%). These results are considered to be because the viscosity of the extraction solvent was lowered and the stirring efficiency was improved by using a mixed solvent.

  On the other hand, when only acetone was used (Comparative Example 1 and Comparative Example 2), when only ethanol was used (Comparative Example 3) and when only ethyl acetate was used (Comparative Example 4), all Compared with the case of using an ionic liquid, the extraction rate was significantly reduced (Comparative Example 1: 0.46%, Comparative Example 2: 0.53%, Comparative Example 3: 0.35, Comparative Example 4: 0.43). %). This result is thought to be due to the fact that when these organic solvents were used, unlike the case of using an ionic liquid, the cellulose of the plant could not be dissolved, so that the extraction rate decreased.

  Thus, according to the above example, by using an ionic liquid and a mixed solvent of an ionic liquid and an organic solvent, at 25 ° C., a maximum of 1.5 times that when only a polar solvent, acetone, is used. It can be seen that the extraction rate can be obtained, and at 50 ° C., the extraction rate can be improved by up to 1.4 times compared to the case of using only the polar solvent acetone.

  Next, the present inventors examined the relationship between the extraction time and the extraction rate for synaropicrin extraction using an ionic liquid. The contents examined by the inventors will be described below.

Example 9
Under the conditions of Example 2, the extraction rates at extraction times of 15 minutes, 30 minutes, 60 minutes, 90 minutes, and 120 minutes were measured. The results are shown in Table 2 below and FIG.

(Example 10)
Under the conditions of Example 3, the extraction rates at extraction times of 15 minutes, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 150 minutes, and 180 minutes were measured. The results are shown in Table 2 below and FIG.

(Comparative Example 5)
Under the conditions of Comparative Example 2, extraction rates at extraction times of 15 minutes, 30 minutes, 60 minutes, 90 minutes, and 120 minutes were measured. The results are shown in Table 2 below and FIG.

  As can be seen from Table 2 and FIG. 3, when only the ionic liquid was used, the extraction rate tended to gradually increase after 60 minutes, and the extraction rate reached 0.62% at the maximum (Example) 9). In addition, when a 1: 1 mixed solvent of ionic liquid and acetone was used, the extraction rate was improved as compared with the case where only the ionic liquid was used, and the extraction rate reached 0.92% at the maximum (implementation). Example 10). On the other hand, when only acetone was used, the extraction rate was not improved between 15 minutes and 120 minutes (Comparative Example 5). From these results, it can be seen that the extraction rate is improved by using the ionic liquid, and that the extraction can be efficiently performed up to 1.9 times higher than the acetone by using the mixed liquid of the ionic liquid and acetone.

  Next, the present inventors can recycle the ionic liquid by regenerating the mixed solvent of 1: 1 and ionic liquid according to Example 3 and Example 10 described above and acetone as the extraction solvent. I investigated whether or not. The contents examined by the inventors will be described below.

(Example 11)
In the same manner as in Example 3, the artichoke leaves were extracted with a 1: 1 mixed solvent of ionic liquid and acetone, and separated into an organic layer and an ionic liquid layer containing artichoke leaves by centrifugation. The organic layer was analyzed by HPLC to calculate the first extraction rate. Next, ethanol (0.4 g) was added to the ionic liquid layer containing artichoke leaves, stirred for 30 seconds, centrifuged for 1 minute, and then filtered to remove the artichoke leaves. After repeating this operation 4 times, the ionic liquid layer was concentrated. Acetone (0.25 g) was added to the obtained ionic liquid layer and reused as a mixed solvent of the ionic liquid and acetone, and extraction was carried out in the same manner as in Example 3. The above operation was repeated 9 times, and the change of the extraction rate from the 1st time to the 9th time was confirmed. The results are shown in FIG.

  As shown in FIG. 4, as a result of examining the reuse of a 1: 1 mixed solvent of ionic liquid and acetone, the extraction rate did not decrease even when the reuse was repeated nine times (first time: 0. 0. 67%, 2nd: 0.75%, 3rd: 0.70%, 4th: 0.62%, 5th: 0.72%, 6th: 0.59%, 7th (Eighth: 0.69%, Ninth: 0.87%). From this result, the mixed solvent of the ionic liquid and the organic solvent according to the above example does not require a complicated regeneration process such as washing with water and liquid separation, and decantation using dilution with an organic solvent and centrifugation. It can be seen that the ionic liquid can be easily regenerated and reused.

  Next, the present inventors examined the extraction solvent, the extraction temperature, and the extraction rate for the extraction of quinicin using an ionic liquid. The contents examined by the inventors will be described below.

(Example 12)
Santori leaves (0.026 g) and ionic liquid (0.5 g) pulverized by a mixer were added to the sample tube and stirred at 25 ° C. for 60 minutes. Thereafter, distilled water (0.214 g) and ethyl acetate (0.714 g) were added and stirred for 5 minutes, followed by centrifugation for 1 minute to separate the extract from the organic layer (ethyl acetate layer). Was recovered by decantation. After repeating the operations of adding ethyl acetate, centrifuging, and separating the organic layer three times, the obtained organic layer was analyzed by reversed-phase high performance liquid chromatography under the following analytical conditions, and an absolute calibration curve method was used. The extraction rate of quinicin was quantified. As a result, the extraction rate of quinicin was 0.56%. The results are shown in Table 3 below and FIG.
(HPLC analysis conditions)
Column: YMC-Pack ODS-AM (150 × 4.6 mm)
Solvent: Measured by the gradient method under the following conditions.
0 min to 5 min; acetonitrile / distilled water = 2: 98
5 min to 30 min; acetonitrile / distilled water = 100: 0
30 min to 40 min; acetonitrile / distilled water = 100: 0
40 min to 45 min; acetonitrile / distilled water = 2: 98
45 min to 55 min; acetonitrile / distilled water = 2: 98
Flow rate: 1.0 mL / min
Column temperature: 40 ° C
UV detection: 228 nm

(Example 13)
The extraction rate of quinicine was measured in the same manner as in Example 12 except that a mixed solvent of ionic liquid (0.25 g) and ethanol (0.25 g) was used instead of ionic liquid (0.5 g). . As a result, the extraction rate of quinicin was 0.66%. The results are shown in Table 3 below and FIG.

(Example 14)
The extraction rate of quinicine was measured in the same manner as in Example 12 except that a mixed solvent of ionic liquid (0.25 g) and acetone (0.25 g) was used instead of ionic liquid (0.5 g). . As a result, the extraction rate of quinicin was 0.67%. The results are shown in Table 3 below and FIG.

(Example 15)
The extraction rate of quinicine was measured in the same manner as in Example 12 except that a mixed solvent of ionic liquid (0.25 g) and ethyl acetate (0.25 g) was used instead of ionic liquid (0.5 g). did. As a result, the extraction rate of quinicin was 0.58%. The results are shown in Table 3 below and FIG.

(Comparative Example 6)
Santori leaves (0.026 g) and ethanol (0.5 g) pulverized by a mixer were added to the sample tube and stirred at 25 ° C. for 60 minutes. Then, after further stirring for 15 minutes, the organic layer (acetone) was separated by centrifuging for 10 minutes. The obtained organic layer was analyzed in the same manner as in Example 12 to quantify the extraction rate of quinicin. As a result, the extraction rate of quinicin was 0.38%. The results are shown in Table 3 below and FIG.

(Comparative Example 7)
The extraction rate of quinicin was measured in the same manner as in Comparative Example 6 except that acetone (0.5 g) was used instead of ethanol (0.5 g). As a result, the extraction rate of quinicin was 0.49%. The results are shown in Table 3 below and FIG.

(Comparative Example 8)
The extraction rate of quinicin was measured in the same manner as in Comparative Example 6 except that ethyl acetate (0.5 g) was used instead of ethanol (0.5 g). As a result, the extraction rate of quinicin was 0.33%. The results are shown in Table 3 below and FIG.

  As can be seen from Table 3 and FIG. 5, when the ionic liquid was used as an extraction solvent, the extraction rate of quinicin was 0.56% (Example 12). When the ionic liquid and the organic solvent were mixed (Example 13: Ethanol, Example 14: Acetone, Example 15: Ethyl acetate), the extraction rate was Example 13: 0.66%. Example 14: Improved to 0.67% and Example 15: 0.58%. This result is considered because the extraction rate was improved as a result of mixing the ionic liquid and the organic solvent to lower the viscosity of the extraction solvent and increase the stirring efficiency.

  In contrast, when only ethanol was used as the extraction solvent (Comparative Example 6), only acetone was used (Comparative Example 7), and only ethyl acetate (Comparative Example 8) was extracted. The rate greatly deteriorated (Comparative Example 6: 0.38%, Comparative Example 7: 0.49%, Comparative Example 8: 0.33%). This result is thought to be because these organic solvents could not dissolve the cellulose of the leaves of Santori and could not extract quinicine efficiently.

  Thus, according to this example, by using a mixed solvent of ionic liquid and acetone, not only can extraction be 1.4 times more efficient than when only acetone is used, It can be seen that efficient extraction using centrifugation and decantation can be realized.

Claims (8)

An extraction step of mixing an extraction solvent containing an ionic liquid capable of dissolving cellulose and a plant containing terpenes, and extracting the terpenes from the plant together with the extraction solvent as an extract,
A step of mixing the extract and an organic solvent capable of dissolving the terpenes, separating the extract and the organic solvent by centrifugation, and collecting the terpenes in the organic solvent. The extraction method of terpenes characterized by these.   The method for extracting terpenes according to claim 1, wherein in the recovery step, the extract, the organic solvent, and water are mixed.   The terpenes according to claim 1 or 2, wherein the extraction solvent is a mixed solvent of the ionic liquid and a polar solvent containing at least one selected from the group consisting of acetones, alcohols, and esters. Extraction method.   The extraction method of terpenes according to any one of claims 1 to 3, wherein in the extraction step, a temperature during the extraction is 15 ° C or higher and 60 ° C or lower.   The terpene extraction method according to claim 3, wherein the mixed solvent has a mass ratio of the ionic liquid to the polar solvent (ionic liquid: polar solvent) in the range of 5: 1 to 1: 5.   Furthermore, after adding the said organic solvent to the extract liquid isolate | separated in the said collection | recovery process and mixing, the said regeneration liquid and the said organic solvent are isolate | separated by centrifugation, The reproduction | regeneration process of regenerating the said ionic liquid is included. The method for extracting terpenes according to any one of claims 1 to 5. The terpene extraction method according to any one of claims 1 to 6, wherein the ionic liquid is represented by the following general formula (1).

(In formula (1), R represents an alkyl group having 1 to 5 carbon atoms.)   The method for extracting terpenes according to any one of claims 1 to 7, wherein the terpenes are sesquiterpenes.

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