Method for isolating and purifying neophytadiene from tobacco leaves, neophytadiene composition, non-combustion heating type flavor inhaler, non-combustion non-heating type flavor inhaler and combustion type flavor inhaler, and methods for producing the same

Abstract

The present invention provides a method that makes it possible to isolate and purify neophytadiene from leaf tobacco with high yield and high purity. The method comprises: a step for adding an aprotic solvent to leaf tobacco to perform an extraction operation and removing extraction residue to obtain an extraction liquid (A1); a step for adding an acidic aqueous solution to the extraction liquid (A1) to perform liquid-liquid extraction, and removing an aqueous layer to obtain an extraction liquid (B1); a step for adding a protic solvent to the extraction liquid (B1) to perform liquid-liquid extraction, and removing a protic solvent layer to obtain an extraction liquid (C1); a step for removing a solvent from the extraction liquid (C1) to obtain dried matter (D1); a step for adding a protic solvent to the dried matter (D1) to obtain a solution (E1); a step for removing precipitate produced in the solution (E1) to obtain a supernatant liquid (F1); a step for replacing the solvent in the supernatant liquid (F1) with an aprotic solvent to obtain a solution (G1); and a step for isolating and purifying neophytadiene from the solution (G1) via normal phase HPLC.

Description

[Technical Field]

[0001] The present invention relates to a method for isolating and purifying neophytadiene from tobacco leaves, a neophytadiene composition, a non-combustion, non-heating type flavor inhaler, a non-combustion, non-heating type flavor inhaler, and a combustion type flavor inhaler, and methods for producing the same. [Background technology]

[0002] Neophytadiene is a terpenoid compound contained in tobacco leaves, and because it correlates with the characteristic aroma of tobacco plants, it has been attracting attention as an important component in the expression of aroma quality. Essential oils containing neophytadiene have been known to have anti-inflammatory effects (Non-Patent Document 1), but in recent years, it has been discovered that neophytadiene itself exerts anti-inflammatory effects, drawing attention (Non-Patent Document 2).

[0003] In addition to tobacco, algae are known to contain neophytadiene among plants (Non-Patent Document 2). It has been reported that neophytadiene is derived from phytol, which is liberated in the rumen by hydrolysis of part of the chlorophyll a and b contained in plants, and it has also been reported that neophytadiene has been detected in milk and beef from cows fed grass containing neophytadiene (Non-Patent Document 3).

[0004] The primary method for obtaining neophytadiene as a simple substance is a synthetic reaction. In the synthetic reaction, chlorophyll is first hydrolyzed under alkaline conditions to produce phytol. Concentrated sulfuric acid is added to the produced phytol and heated at 170°C, causing an elimination reaction to produce neophytadiene (Patent Document 1).

[0005] On the other hand, when neophytadiene is isolated from a mixture of plants, etc., it is refined as an essential oil containing other substances, and there have been few reports on the isolation and purification of neophytadiene itself. For example, Non-Patent Document 4 describes a process for isolating and purifying neophytadiene from tobacco, in which neophytadiene is extracted from flue-cured tobacco with a hexane solvent, the hexane layer is eluted using silicic acid column chromatography, the solvent is replaced with acetone, saturated hydrocarbons are crystallized and removed, and then the elution is performed using alumina chromatography. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Special Publication No. 2012-529893 [Non-patent literature]

[0007] [Non-Patent Document 1] Adeosun TE, Ogunwande IA, Avoseh ON, Raji IP, Lawal OA. Composition and Anti-inflammatory Activity of Essential Oil of Jatropha curcas. Nat Prod Commun. 2017 Mar;12(3):439-440. PMID: 30549905. [Non-patent document 2] Bhardwaj, M., Sali, VK, Mani, S. et al. Neophytadiene from Turbinaria ornata Suppresses LPS-Induced Inflammatory Response in RAW 264.7 Macrophages and Sprague Dawley Rats. Inflammation 43, 937-950 (2020) [Non-patent document 3] Study on the characteristics of volatile compounds in milk produced by pasture-raised cows and the establishment of traceability using volatile compounds in milk as markers, Yasuko Ueda, Hokkaido University, Ph.D. (Agriculture) No. 6987 (2016) [Non-patent document 4] Flue-cured Tobacco. II. Neophytadiene Summary of the Invention [Problem to be solved by the invention]

[0008] However, the method described in Non-Patent Document 4 does not allow for the isolation and purification of neophytadiene in high yield and with high purity. Considering the expansion of effective utilization of neophytadiene, it is desirable to develop a method for isolating and purifying neophytadiene from tobacco leaves in high yield and with high purity.

[0009] The present invention aims to provide a method for isolating and purifying neophytadiene from tobacco leaves in high yield and with high purity, a neophytadiene composition obtained by the method, and a non-combustion, non-heating, flavor inhaler, and combustion-type flavor inhaler each containing the neophytadiene composition. [Means for solving the problem]

[0010] The present invention includes the following embodiments.

[0011] [1] A method for isolating and purifying neophytadiene from tobacco leaves, comprising: a step of adding an aprotic solvent to tobacco leaves to perform an extraction operation, and removing the extraction residue to obtain an extract (A1); a step of adding an acidic aqueous solution to the extract (A1) to perform liquid-liquid extraction, and removing the aqueous layer to obtain an extract (B1); a step of adding a protic solvent to the extract (B1) to perform liquid-liquid extraction, and removing the protic solvent layer to obtain an extract (C1); a step of removing the solvent from the extract (C1) to obtain a dry product (D1); a step of adding a protic solvent to the dried product (D1) to dissolve the dried product (D1) and obtain a solution (E1); a step of removing a precipitate formed in the solution (E1) to obtain a supernatant (F1); a step of replacing the solvent of the supernatant (F1) with an aprotic solvent to obtain a solution (G1); isolating and purifying neophytadiene from the solution (G1) by normal phase high performance liquid chromatography; A method comprising:

[0012] [2] A method for isolating and purifying neophytadiene from tobacco leaves, comprising: a step of adding an aprotic solvent to tobacco leaves to perform an extraction operation, and removing the extraction residue to obtain an extract (A2); a step of adding an acidic aqueous solution to the extract (A2) to perform liquid-liquid extraction, and removing the aqueous layer to obtain an extract (B2); a step of removing the solvent from the extract (B2) to obtain a dry product (C2); a step of adding a protic solvent to the dried product (C2) to dissolve the dried product (C2) and obtain a solution (D2); removing a precipitate formed in the solution (D2) to obtain a supernatant (E2); a step of adding an aprotic solvent to the supernatant (E2) to perform liquid-liquid extraction, and removing the protic solvent layer to obtain a solution (F2); isolating and purifying neophytadiene from the solution (F2) by normal phase high performance liquid chromatography; A method comprising:

[0013] [3] The method according to [1] or [2], wherein the pH of the acidic aqueous solution is 3 or less.

[0014] [4] The method according to any one of [1] to [3], wherein the aprotic solvent is at least one solvent selected from the group consisting of hexane, heptane, and ethyl acetate.

[0015] [5] The method according to any one of [1] to [4], wherein the protic solvent is methanol.

[0016] [6] The method according to any one of [1] to [5], wherein the tobacco leaf is at least one of flue-cured and burley.

[0017] [7] The method according to [1], wherein the step of obtaining the solution (G1) is a step of adding an aprotic solvent to the supernatant (F1) to perform liquid-liquid extraction, and removing the protic solvent layer to obtain the solution (G1).

[0018] [8] The method according to [1], wherein the step of obtaining the solution (G1) is a step of removing the solvent from the supernatant (F1) and then adding an aprotic solvent to obtain the solution (G1).

[0019] [9] The method according to any one of [1] to [8], wherein the neophytadiene composition obtained by the isolation and purification contains neophytadiene in an amount of 95 mass% or more.

[0020]

[10] A step of isolating and purifying neophytadiene from tobacco leaves by the method according to any one of [1] to [9] to obtain a neophytadiene composition; adding the neophytadiene composition to a substrate; a step of manufacturing a non-combustion heating type flavor inhaler including the base material; A method for manufacturing a non-combustion heating type flavor inhaler, comprising:

[0021]

[11] A step of isolating and purifying neophytadiene from tobacco leaves by the method according to any one of [1] to [9] to obtain a neophytadiene composition; adding the neophytadiene composition to a substrate; a step of manufacturing a non-combustion, non-heating flavor inhaler comprising the base material; A method for manufacturing a non-burning, non-heating flavor inhaler, comprising:

[0022]

[12] A step of isolating and purifying neophytadiene from tobacco leaves by the method according to any one of [1] to [9] to obtain a neophytadiene composition; adding the neophytadiene composition to a substrate; manufacturing a combustion-type flavor inhaler comprising the substrate; A method for manufacturing a combustion type flavor inhaler, comprising:

[0023]

[13] A neophytadiene composition containing 95% by mass or more of neophytadiene, obtained by isolating and purifying neophytadiene from tobacco leaves by the method according to any one of [1] to [9].

[0024]

[14] A non-combustion heating type flavor inhaler containing the neophytadiene composition according to

[13] .

[0025]

[15] A non-combustion, non-heating type flavor inhaler containing the neophytadiene composition according to

[13] .

[0026]

[16] A combustion-type flavor inhaler containing the neophytadiene composition according to

[13] . [Effects of the Invention]

[0027] According to the present invention, there are provided a method for isolating and purifying neophytadiene from tobacco leaves in high yield and with high purity, a neophytadiene composition obtained by the method, a non-combustion and heating type flavor inhaler, a non-combustion and non-heating type flavor inhaler, and a combustion type flavor inhaler each containing the neophytadiene composition. [Brief explanation of the drawings]

[0028] [Figure 1] 1 is a flowchart illustrating an example of a method according to a first embodiment of the present invention. [Figure 2] 10 is a flowchart illustrating an example of a method according to a second embodiment of the present invention. [Figure 3] 1 is a schematic diagram illustrating an example of a non-combustion heating type flavor inhaler according to an embodiment of the present invention. FIG. [Figure 4]1 is a schematic diagram showing an example of a non-combustion heating type flavor inhalation system according to an embodiment of the present invention. [Figure 5] 1 is a schematic diagram illustrating an example of a non-combustion, non-heating type flavor inhaler according to an embodiment of the present invention. FIG. [Figure 6] 1 is a schematic diagram showing an example of a tobacco capsule of a non-combustion, non-heating type flavor inhaler according to an embodiment of the present invention. FIG. [Figure 7] FIG. 2 is a schematic diagram showing an example of a power supply unit of a non-combustion and non-heating type flavor inhaler according to the present embodiment. [Figure 8] 1 is a schematic diagram showing an example of a cartridge of a non-combustion, non-heating type flavor inhaler according to an embodiment of the present invention. FIG. [Figure 9] 1 is a schematic diagram showing an example of a cartridge of a non-combustion, non-heating type flavor inhaler according to an embodiment of the present invention. FIG. [Figure 10] 1 is a schematic diagram illustrating an example of a combustion-type flavor inhaler according to an embodiment of the present invention. [Figure 11] 1 is a total ion chromatogram obtained by GC / MS analysis of an n-hexane solution in Reference Example 2. [Figure 12] 1 is a total ion chromatogram obtained by GC / MS analysis of an ethyl acetate solution in Reference Example 2. [Figure 13] 1 is a total ion chromatogram obtained by GC / MS analysis of an ethanol solution in Reference Example 2. [Figure 14] 1 is a total ion chromatogram obtained by GC / MS analysis of a methanol solution in Reference Example 2. [Figure 15] 1 is a total ion chromatogram obtained by GC / MS analysis of the solution obtained in step B1 of Example 1. [Figure 16] 1 is a total ion chromatogram obtained by GC / MS analysis of the hexane solution obtained in step C1 of Example 1. [Figure 17] 1 is a total ion chromatogram obtained by GC / MS analysis of the methanol solution obtained in step C1 of Example 1. [Figure 18]1 is a total ion chromatogram obtained by GC / MS analysis of the supernatant obtained in step F1 of Example 1. [Figure 19] 1 is a chromatogram of neophytadiene fractionated by HPLC according to step H1 of Example 1. [Figure 20] 1 is a total ion chromatogram obtained by GC / MS analysis of the neophytadiene composition solution obtained in Example 1. DETAILED DESCRIPTION OF THE INVENTION

[0029] [Method for isolating and purifying neophytadiene from tobacco leaves] First Embodiment The method for isolating and purifying neophytadiene from tobacco leaves according to this embodiment includes the following steps. a step of adding an aprotic solvent to tobacco leaves to perform an extraction operation, and removing the extraction residue to obtain an extract (A1) (hereinafter also referred to as step A1); a step of adding an acidic aqueous solution to the extract (A1) to perform liquid-liquid extraction and removing the aqueous layer to obtain an extract (B1) (hereinafter also referred to as step B1); a step of adding a protic solvent to the extract (B1) to perform liquid-liquid extraction, and removing the protic solvent layer to obtain an extract (C1) (hereinafter also referred to as step C1); a step of removing the solvent from the extract (C1) to obtain a dry product (D1) (hereinafter also referred to as step D1); a step of adding a protic solvent to the dried product (D1) to dissolve the dried product (D1) and obtain a solution (E1) (hereinafter also referred to as step E1); a step of removing the precipitate formed in the solution (E1) to obtain a supernatant (F1) (hereinafter also referred to as step F1); A step of replacing the solvent in the supernatant (F1) with an aprotic solvent to obtain a solution (G1) (hereinafter also referred to as step G1); and A step of isolating and purifying neophytadiene from the solution (G1) by normal phase high performance liquid chromatography (hereinafter also referred to as step H1).

[0030] The method according to the present embodiment, which includes steps A1 to H1, enables the isolation and purification of neophytadiene from tobacco leaves in high yield and purity. Because normal-phase high-performance liquid chromatography (HPLC) typically cannot completely separate neophytadiene from nicotine, it is difficult to isolate and purify neophytadiene in high yield and purity. However, in the method according to the present embodiment, step B1 is first performed on the extract obtained from tobacco leaves in step A1, and alkaloids, primarily consisting of nicotine, are separated from the extract by adding an acidic aqueous solution and removing the aqueous layer. This first removes nicotine and other components, thereby efficiently improving the yield and purity of the isolated and purified neophytadiene. Furthermore, the solvent in the extract (C1) is intentionally removed in step D1 to obtain a dried product (D1). After this, a protic solvent is added in step E1 to redissolve the dried product (D1), and the precipitate (higher hydrocarbons) formed in the resulting solution (E1) is removed in step F1, thereby efficiently separating the higher hydrocarbons. This allows the yield and purity of the isolated and purified neophytadiene to be more efficiently improved. Details of steps A1 to H1 are described below, but the method according to this embodiment may include steps other than steps A1 to H1. A flowchart of an example of the method according to this embodiment is shown in Figure 1.

[0031] (Process A1) In this step, an aprotic solvent is added to tobacco leaves to perform an extraction operation, and the extraction residue is removed to obtain an extract (A1). Through this step, active ingredients, including neophytadiene, contained in tobacco leaves can be extracted into the extract (A1).

[0032] The type of leaf tobacco is not particularly limited, but examples include flue-cured, burley, oriental, native, other Nicotiana tabacum varieties, and Nicotiana rustica varieties. One or more of these leaf tobacco varieties may be used. Among these, at least one type of leaf tobacco selected from the group consisting of flue-cured and burley varieties is preferred, with flue-cured being more preferred, in terms of containing a large amount of neophytadiene. The form of the leaf tobacco is not particularly limited, but finely powdered leaf tobacco is preferred in terms of improving extraction efficiency. Solid waste discarded from the leaf tobacco expansion process and finely powdered leaf tobacco discarded from leaf tobacco raw material factories may also be used as the leaf tobacco.

[0033] The aprotic solvent used as the extraction solvent is preferably at least one solvent selected from the group consisting of hexane, heptane, and ethyl acetate, with hexane being more preferred, from the viewpoint of enabling more selective extraction of neophytadiene. The amount of aprotic solvent added is preferably 500 to 2,000 parts by mass, more preferably 750 to 1,250 parts by mass, per 100 parts by mass of tobacco leaves. The extraction procedure can be carried out by stirring the aprotic solvent containing tobacco leaves, for example. The extraction temperature is preferably 15 to 40°C, and the extraction time is preferably 1 to 6 hours. The extraction residue can be removed by separating the extract (A1) from the extraction residue using, for example, a stainless steel mesh.

[0034] (Process B1) In this step, an acidic aqueous solution is added to the extract (A1) obtained in step A1 to perform liquid-liquid extraction, and the aqueous layer is removed to obtain extract (B1). In this step, alkaloids, primarily nicotine, migrate to the aqueous layer, while hydrophobic active ingredients in tobacco leaves, such as neophytadiene, remain in the organic layer, allowing for the selective separation and removal of alkaloids, primarily nicotine.

[0035] In this embodiment, an acidic aqueous solution refers to an aqueous solution having a pH of less than 7. The acidic aqueous solution may be an aqueous solution containing a pH adjuster such as sulfuric acid or hydrochloric acid. The pH of the acidic aqueous solution is preferably 3 or less, more preferably 1 to 3, from the viewpoint of facilitating the migration of alkaloids, typified by nicotine, into the aqueous layer. In this specification, the pH of the aqueous solution is a value measured with a pH meter (for example, a desktop pH meter (trade name), manufactured by HORIBA).

[0036] The amount of acidic aqueous solution added depends on the pH of the acidic aqueous solution, but the volume ratio of the extract (A1) to the acidic aqueous solution (extract (A1):acidic aqueous solution) is preferably 50 to 500:100, more preferably 200 to 400:100. The liquid-liquid extraction operation can be performed by shaking the mixed solution, etc. The extraction temperature is preferably 15 to 40°C, and the extraction time is preferably 0.5 to 3 hours. A salt such as sodium chloride may be added to the mixed solution during liquid-liquid extraction. A desiccant such as anhydrous sodium sulfate may be added to the extract (B1) obtained by separating and removing the aqueous layer, to dehydrate the extract (B1).

[0037] (Process C1) In this step, a protic solvent is added to the extract (B1) obtained in step B1 to perform liquid-liquid extraction, and the protic solvent layer is removed to obtain extract (C1). In this step, the leaf surface resin components of tobacco leaf containing cembratriene diol (CBT) as a main component are transferred to the protic solvent layer, while neophytadiene and higher hydrocarbons are transferred to the aprotic solvent layer, allowing the leaf surface resin components of tobacco leaf containing CBT as a main component to be selectively separated and removed.

[0038] As the protic solvent, methanol is preferred from the viewpoint of being able to more selectively extract and remove leaf surface resin components from tobacco leaves, which are primarily composed of CBT. The amount of protic solvent added is preferably such that the volume ratio of the extract (B1) to the protic solvent (extract (B1):protic solvent) is 100:15-100, more preferably 100:40-60. Liquid-liquid extraction can be performed by shaking the mixture, etc. The extraction temperature is preferably 15-40°C, and the extraction time is preferably 3-8 hours. Furthermore, from the viewpoint of further removing leaf surface resin components from tobacco leaves, which are primarily composed of CBT, this step may be repeated two or more times.

[0039] (Process D1) In this step, the solvent of the extract (C1) obtained in the step C1 is removed to obtain a dry product (D1). The method for removing the solvent (aprotic solvent) of the extract (C1) is not particularly limited, but the solvent can be removed from the extract (C1) under reduced pressure, for example.

[0040] (Process E1) In this step, a protic solvent is added to the dried product (D1) obtained in step D1 to dissolve the dried product (D1) and obtain a solution (E1). Methanol is preferred as the protic solvent, as it is less soluble in higher hydrocarbons and more soluble in neophytadiene. The amount of the protic solvent added is preferably 750 to 2,500 parts by mass, more preferably 1,000 to 1,500 parts by mass, per 100 parts by mass of the dried product (D1). The method for dissolving the dried product (D1) in the protic solvent is not particularly limited, but the dried product (D1) can be dispersed and dissolved, for example, by subjecting the solution to ultrasonic waves. It is not necessary for the dried product (D1) to be completely dissolved in the protic solvent; it is sufficient for at least a portion of the dried product to be dissolved.

[0041] (Process F1) In this step, the precipitate formed in the solution (E1) obtained in the step E1 is removed to obtain a supernatant (F1). This step makes it possible to selectively separate and remove higher hydrocarbons as precipitates. The precipitate can be formed by allowing the solution (E1) to stand. The standing can be carried out, for example, at 0 to 30°C for 1 to 12 hours.

[0042] (Process G1) In this step, the solvent of the supernatant (F1) obtained in the step F1 is replaced with an aprotic solvent to obtain a solution (G1). The method for this step is not particularly limited as long as it can replace the solvent (protic solvent) of the supernatant (F1) with an aprotic solvent. For example, the solution (G1) can be obtained by adding an aprotic solvent to the supernatant (F1) and performing liquid-liquid extraction, followed by removing the protic solvent layer. Alternatively, the solution (G1) can be obtained by removing the solvent from the supernatant (F1) and then adding an aprotic solvent.

[0043] (Process H1) In this step, neophytadiene is isolated and purified from the solution (G1) obtained in step G1 by normal-phase high-performance liquid chromatography (normal-phase HPLC). The HPLC used is not particularly limited, but a commercially available product such as 1260 Infinity (trade name, manufactured by Agilent Technologies) can be used. When using HPLC, the separation procedure can be performed, for example, under the following conditions. Column: YMC-Pack SIL / S-5μm / 12nm ·Mobile phase: Hex(2min)→Hex:AcOEt(95:5)(1min)→Hex(12min) ·Flow rate: 7mL / min Detector: Diode array detector (220 nm) Sample injection volume: 500 μL

[0044] After separation under the above conditions, the signal detected at 220 nm is fractionated in a fraction collector, the fractionated fractions are collected, and the obtained sample is concentrated and dried to isolate and purify neophytadiene. The method according to this embodiment can obtain neophytadiene in high yield, and the neophytadiene composition obtained by isolation and purification contains neophytadiene in an amount of 95% by mass or more.

[0045] <Second embodiment> The method for isolating and purifying neophytadiene from tobacco leaves according to this embodiment includes the following steps. a step of adding an aprotic solvent to tobacco leaves to perform an extraction operation, and removing the extraction residue to obtain an extract (A2) (hereinafter also referred to as step A2); a step of adding an acidic aqueous solution to the extract (A2) to perform liquid-liquid extraction and removing the aqueous layer to obtain an extract (B2) (hereinafter also referred to as step B2); a step of removing the solvent from the extract (B2) to obtain a dry product (C2) (hereinafter also referred to as step C2); a step of adding a protic solvent to the dried product (C2) to dissolve the dried product (C2) and obtain a solution (D2) (hereinafter also referred to as step D2); a step of removing the precipitate formed in the solution (D2) to obtain a supernatant (E2) (hereinafter also referred to as step E2); a step of adding an aprotic solvent to the supernatant (E2) to perform liquid-liquid extraction, and removing the protic solvent layer to obtain a solution (F2) (hereinafter also referred to as step F2); a step of isolating and purifying neophytadiene from the solution (F2) by normal phase high performance liquid chromatography (hereinafter also referred to as step G2).

[0046] The method according to the present embodiment, including steps A2 to G2, can isolate and purify neophytadiene from tobacco leaves with high purity. In particular, the method according to the present embodiment first performs step B2 on the extract obtained from tobacco leaves in step A2, adding an acidic aqueous solution and removing the aqueous layer to separate alkaloids, primarily nicotine, contained in the extract. This process removes nicotine and other components first, thereby efficiently improving the yield and purity of the isolated and purified neophytadiene. Furthermore, the solvent from the extract (B2) is intentionally removed in step C2 to obtain a dried product (C2). Then, a protic solvent is added in step D2 to redissolve the dried product (C2). The precipitate (higher hydrocarbons) formed in the resulting solution (D2) is then removed in step E2, allowing for efficient separation of the higher hydrocarbons. This process can efficiently improve the yield and purity of the isolated and purified neophytadiene. Details of steps A2 to G2 are described below, but the method according to the present embodiment may also include steps other than steps A2 to G2. A flowchart of an example of a method according to this embodiment is shown in FIG.

[0047] (Process A2, B2) Steps A2 and B2 can be performed in the same manner as steps A1 and B1 in the first embodiment.

[0048] (Process C2) In this step, the solvent is removed from the extract (B2) obtained in step B2 to obtain a dry product (C2). The method for removing the solvent (aprotic solvent) from the extract (B2) is not particularly limited, but the solvent can be removed from the extract (B2) under reduced pressure, for example.

[0049] (Process D2) In this step, a protic solvent is added to the dried product (C2) obtained in step C2 to dissolve the dried product (C2) and obtain a solution (D2). Methanol is preferred as the protic solvent, as it is less soluble in higher hydrocarbons and more soluble in neophytadiene. The amount of protic solvent added is preferably 500 to 2,500 parts by mass, more preferably 750 to 1,500 parts by mass, per 100 parts by mass of the dried product (C2). The method for dissolving the dried product (C2) in the protic solvent is not particularly limited, but the dried product (C2) can be dispersed and dissolved, for example, by subjecting the solution to ultrasonic waves. It is not necessary for the dried product (C2) to be completely dissolved in the protic solvent; it is sufficient for at least a portion of the dried product to be dissolved.

[0050] (Process E2) In this step, the precipitate formed in the solution (D2) obtained in the step D2 is removed to obtain a supernatant (E2). This step makes it possible to selectively separate and remove higher hydrocarbons as precipitates. The precipitate can be formed by allowing the solution (D2) to stand. The standing can be carried out, for example, at 15 to 40°C for 3 to 8 hours.

[0051] (Process F2) In this step, an aprotic solvent is added to the supernatant (E2) obtained in step E2 to perform liquid-liquid extraction, and the protic solvent layer is removed to obtain a solution (F2). In this step, the leaf surface resin component of tobacco leaf containing cembratriene diol (CBT) as a main component migrates to the protic solvent layer, and neophytadiene migrates to the aprotic solvent layer, so that the leaf surface resin component of tobacco leaf containing CBT as a main component can be selectively separated and removed.

[0052] The aprotic solvent is preferably at least one solvent selected from the group consisting of hexane, heptane, and ethyl acetate, and more preferably hexane, from the viewpoint of more selectively extracting neophytadiene. The volume ratio of the supernatant (E2) to the aprotic solvent (supernatant (E2): aprotic solvent) is preferably 15 to 100:100, more preferably 40 to 60:100. Liquid-liquid extraction can be performed by shaking the mixture. The extraction temperature is preferably 0 to 30°C, and the extraction time is preferably 3 to 12 hours. To further remove the surface resin component of tobacco leaf, which is primarily composed of CBT, this step may be repeated two or more times.

[0053] (Process G2) Step G2 can be carried out in the same manner as Step H1 in Embodiment 1. In the method of this embodiment, neophytadiene can be obtained in high yield, as in Embodiment 1, and the neophytadiene composition obtained by isolation and purification contains 95 mass % or more of neophytadiene.

[0054] [Neophytadiene composition] The neophytadiene composition according to this embodiment is obtained by the method for isolating and purifying neophytadiene from tobacco leaves according to the embodiment described above, and contains 95% by mass or more of neophytadiene. The neophytadiene composition preferably contains 97% by mass or more, and more preferably 99% by mass or more, of neophytadiene. The content (purity) of neophytadiene in the neophytadiene composition is a value measured by GC. Components other than neophytadiene contained in the neophytadiene composition obtained by the method according to this embodiment are mainly various components derived from tobacco leaves, and these are difficult to identify because they are present in only trace amounts.

[0055] [Non-combustion heating type flavor inhaler and its manufacturing method] A method for producing a non-combustion heating type flavor inhaler according to this embodiment includes the steps of isolating and purifying neophytadiene from tobacco leaves to obtain a neophytadiene composition by the method according to this embodiment, adding the neophytadiene composition to a substrate, and producing a non-combustion heating type flavor inhaler including the substrate. Furthermore, the non-combustion heating type flavor inhaler according to this embodiment includes the neophytadiene composition according to this embodiment. The method for producing a non-combustion heating type flavor inhaler according to this embodiment obtains the neophytadiene composition by the method according to this embodiment, thereby enabling efficient isolation and purification of neophytadiene from tobacco leaves and efficient production of a non-combustion heating type flavor inhaler that can provide a good flavor. Furthermore, the non-combustion heating type flavor inhaler according to this embodiment can provide a good flavor because it includes the neophytadiene composition obtained by the method according to this embodiment.

[0056] (base material) In the method for producing a non-combustion heating-type flavor inhaler according to the present embodiment, the base material to which the neophytadiene composition is added may be a tobacco material, such as a tobacco sheet, tobacco shreds, cigarette paper, or polysaccharide sheet.

[0057] (1) Tobacco sheets A tobacco sheet is a sheet obtained by molding a composition containing aged tobacco leaves. The aged tobacco leaves used for the tobacco sheet are not particularly limited, but examples include those that have been deboned and separated into lamina and midrib. Aged tobacco leaves refer to tobacco leaves that have undergone processes such as curing and long-term storage in a warehouse or the like. In this embodiment, "sheet" refers to a material having a pair of approximately parallel main and side surfaces. Tobacco sheets can be molded by known methods such as papermaking, casting, and rolling. Details of various tobacco sheets molded by these methods are disclosed in "Encyclopedia of Tobacco," Tobacco Research Center, March 31, 2009. The manner in which the neophytadiene composition according to this embodiment is added to a tobacco sheet is not limited.

[0058] For example, the neophytadiene composition according to the present embodiment may be dissolved in a solvent to prepare a tobacco flavoring agent solution, which may then be sprayed or impregnated onto a completed tobacco sheet. Alternatively, the neophytadiene composition according to the present embodiment may be added when the tobacco sheet is molded. For example, in a papermaking method, water-soluble components are extracted from aged tobacco leaves and separated into an aqueous extract and a residue, a mixture of the fibrous residue and pulp is made into paper, and a concentrated aqueous extract is added to the papersheet. The neophytadiene composition according to the present embodiment can be added to the aqueous extract. In a casting method, water, pulp, a binder, and ground aged tobacco are mixed to form a mixture, which is then cast. The neophytadiene composition according to the present embodiment can be added to this mixture. In a rolling method, water, pulp, a binder, and ground aged tobacco are mixed to form a mixture, which is then fed into a plurality of rolling rollers and rolled. The neophytadiene composition according to the present embodiment can be added to this mixture.

[0059] Furthermore, as described in WO 2014 / 104078, a nonwoven tobacco sheet can be obtained by mixing ground aged tobacco with a binder to form a mixture, sandwiching the mixture between nonwoven fabrics, and molding the laminate into a specific shape by heat welding. In this method, the neophytadiene composition according to the present embodiment can be added to the mixture.

[0060] The tobacco sheet may contain an aerosol-generating substrate. The type of aerosol-generating substrate is not particularly limited, and extracts from various natural products or their components can be selected depending on the application. Specific examples of aerosol-generating substrates include polyhydric alcohols such as glycerin, propylene glycol, sorbitol, xylitol, and erythritol, triacetin, 1,3-butanediol, and mixtures thereof. The content of the aerosol-generating substrate can be adjusted to various amounts depending on the form in which it is used in the tobacco product. For example, when the tobacco sheet contains an aerosol-generating substrate, the content thereof is typically 5% by mass or more, preferably 10% by mass or more, and more preferably 15% by mass or more, and typically 50% by mass or less, preferably 40% by mass or less, and more preferably 25% by mass or less, relative to the total mass of the tobacco sheet, from the perspective of achieving a good flavor.

[0061] (2) Tobacco cuttings Examples of tobacco shreds include aged tobacco leaves shredded to a predetermined size, the aforementioned tobacco sheets shredded to a predetermined size, and mixtures thereof. The size is not limited, and examples include shreds of 0.5 to 2.0 mm in width and 3 to 10 mm in length. Tobacco shreds of this size are preferred for filling a filler, as described below. Other examples of tobacco shreds include strand-type shreds obtained by shredding processed tobacco leaves to a width of 0.5 to 2.0 mm and a length longer than the aforementioned tobacco shreds, preferably approximately the same length as cigarette paper. The neophytadiene composition according to this embodiment may be added to tobacco shreds or to raw materials before shredding.

[0062] The tobacco shreds may contain the aerosol-generating base material. When the aerosol-generating base material is contained in the tobacco shreds, the content thereof is usually 5% by mass or more, preferably 10% by mass or more, and more preferably 15% by mass or more, relative to the mass of the tobacco shreds, from the viewpoints of generating a sufficient amount of aerosol and obtaining a good flavor, and is usually 50% by mass or less, preferably 40% by mass or less, and more preferably 25% by mass or less.

[0063] (3) Rolling paper The neophytadiene composition according to the present embodiment is dissolved in a solvent to prepare a tobacco flavoring solution, which is then sprayed onto and impregnated into cigarette paper, thereby preparing cigarette paper containing the neophytadiene composition. Examples of cigarette paper include those primarily composed of pulp. Pulp may be made from wood pulp, such as softwood pulp or hardwood pulp, or may be made by blending non-wood pulp, such as flax pulp, hemp pulp, sisal pulp, or esparto, which are commonly used in cigarette paper for tobacco articles. These pulps may be used alone or in combination of multiple types in any ratio. The cigarette paper may be composed of one sheet or multiple sheets. The cigarette paper may be used to wrap tobacco raw materials, such as tobacco shreds, and may also be used as a material (e.g., tipping paper) for wrapping the wrapped material together with other components, such as a cooling element or a filter element. As the pulp, chemical pulp produced by the kraft cooking method, acidic, neutral or alkaline sulfite cooking method, soda salt cooking method, etc., ground pulp, chemi-ground pulp, thermomechanical pulp, etc. can be used.

[0064] (4) Polysaccharide sheet A polysaccharide sheet is a sheet primarily composed of polysaccharides, and the neophytadiene composition of the present embodiment can be incorporated into the polysaccharide sheet. A non-combustion, heating-type flavor inhaler using a polysaccharide sheet containing the neophytadiene composition of the present embodiment can release a favorable flavor. Examples of polysaccharides include carrageenan, agar, gellan gum, tamarind gum, psyllium seed gum, konjac glucomannan, carrageenan, locust bean gum, guar gum, agar, xanthan gum, gellan gum, tamarind gum, tara gum, konjac glucomannan, starch, cassia gum, and psyllium seed gum.

[0065] The polysaccharide sheet containing the neophytadiene composition according to the present embodiment can be used in a non-combustion heating type flavor inhaler, as well as in a non-combustion non-heating type flavor inhaler and a combustion type flavor inhaler, which will be described later. In the combustion type flavor inhaler, for example, a polysaccharide sheet such as that disclosed in Japanese Patent No. 5481574 can be used. In this embodiment, the content of the neophytadiene composition according to the present embodiment can be 0.01 to 30 mass% of the sheet. The polysaccharide sheet can be prepared by mixing a polysaccharide with water, heating the mixture to prepare an aqueous solution of the polysaccharide, adding a flavor and an emulsifier to the aqueous solution, and kneading and emulsifying the mixture. Known emulsifiers can be used.

[0066] In an embodiment of the non-combustion heating type flavor inhaler, a polysaccharide sheet such as that described in PCT / JP2019 / 20136 can be used. In this embodiment, it is particularly preferable to use agar as the polysaccharide. The content of agar relative to the sheet is preferably 10 to 50% by mass, more preferably 15 to 45% by mass. Furthermore, the neophytadiene composition according to this embodiment in the polysaccharide sheet can be 0.01 to 30% by mass relative to the sheet.

[0067] In this embodiment, it is preferable to use a sugar compound selected from the group consisting of sugars and sugar alcohols. Examples of "sugars" include glucose, sucrose, fructose, xylose, galactose, mannose, maltose, trehalose, lactose, and raffinose. Examples of "sugar alcohols" include sorbitol, an alcohol obtained by reducing the carbonyl group of a sugar to a hydroxyl group. The content of this compound relative to the mass of agar is preferably 10% by mass or more, more preferably 10 to 500% by mass, even more preferably 10 to 300% by mass, and even more preferably 10 to 200% by mass. In addition, in this embodiment, it is preferable to use an emulsifier. Known emulsifiers can be used, and the content is preferably 0.5 to 10% by mass, more preferably 1.0 to 8.0% by mass, relative to the mass of agar.

[0068] The polysaccharide sheet in this embodiment can be produced by kneading raw materials including agar, a sugar compound, a flavoring, and an emulsifier in water to prepare a raw material slurry, spreading the raw material slurry on a substrate, and drying it.

[0069] (Non-combustion heating type flavor inhaler) In this embodiment, a "flavor inhaler" refers to an item with which a user inhales flavors. Flavor inhalers are broadly classified into "combustion-type flavor inhalers" that generate flavors through combustion and "non-combustion-type flavor inhalers" that generate flavors without combustion. Non-combustion-type flavor inhalers are further broadly classified into "non-combustion-heating-type flavor inhalers" that generate flavors through heating and "non-combustion-non-heating-type flavor inhalers" that generate flavors without heating. A combination of a device for generating aerosols (such as a heating device or an atomizing device) and a non-combustion-heating-type flavor inhaler is also referred to as a non-combustion-heating-type flavor inhalation system.

[0070] FIG. 3 shows one embodiment of a non-combustion heating type flavor inhaler according to this embodiment. As shown in FIG. 3, the non-combustion heating type flavor inhaler 20 includes a tobacco rod portion 20A, a cylindrical cooling portion 20B having perforations on its circumference, and a filter portion 20C. The non-combustion heating type flavor inhaler 20 may include other components. The axial length of the non-combustion heating type flavor inhaler 20 is not limited, but is preferably 40 to 90 mm, more preferably 50 to 75 mm, and even more preferably 50 to 60 mm. The circumferential length of the non-combustion heating type flavor inhaler 20 is preferably 16 to 25 mm, more preferably 20 to 24 mm, and even more preferably 21 to 23 mm. For example, the tobacco rod portion 20A may be 20 mm long, the cooling portion 20B may be 20 mm long, and the filter portion 20C may be 7 mm long. The lengths of these individual components may be appropriately changed depending on manufacturing suitability, required quality, and the like. Although FIG. 3 shows an embodiment in which the first segment 25 is arranged, it is also possible to arrange only the second segment 26 downstream of the cooling section 20B without the first segment 25 being arranged.

[0071] (1) Tobacco rod part 20A The tobacco rod portion 20A can use tobacco shreds or a tobacco sheet containing the neophytadiene composition according to this embodiment as the tobacco filler 21. The method for filling the tobacco filler 21 into the cigarette paper 22 is not particularly limited. For example, the tobacco filler 21 may be wrapped in the cigarette paper 22, or the tobacco filler 21 may be filled into a tubular cigarette paper 22. When the tobacco has a longitudinal direction, such as a rectangular shape, the tobacco may be filled so that the longitudinal direction is in an unspecified direction within the cigarette paper 22, or may be aligned along the axial direction of the tobacco rod portion 20A or perpendicular thereto. Alternatively, the cigarette paper 22 can be the aforementioned cigarette paper containing the neophytadiene composition according to this embodiment. When the tobacco rod portion 20A is heated, the tobacco components, aerosol-generating substrate, and water contained in the tobacco filler 21 vaporize and are available for inhalation.

[0072] (2) Cooling section 20B The cooling section 20B is preferably constructed of a cylindrical member. The cylindrical member may be, for example, a cardboard tube 23 formed by processing cardboard into a cylindrical shape. The cooling section 20B may also be formed from a sheet of thin material that is wrinkled and then pleated, gathered, or folded to form a channel. Examples of such a material include sheet materials selected from the group consisting of polyethylene, polypropylene, polyvinyl chloride, polyethylene terephthalate, polylactic acid, cellulose acetate, and aluminum foil. The total surface area of the cooling section 20B is appropriately adjusted taking into account cooling efficiency, but may be, for example, 300 to 1000 mm 2 / mm. The cooling section 20B is preferably provided with perforations 24. The presence of the perforations 24 allows outside air to be introduced into the cooling section 20B during puffing. As a result, the vaporized components of the aerosol generated by heating the tobacco rod section 20A come into contact with the outside air, and as their temperature drops, they are liquefied, forming an aerosol. The diameter (distance across) of the perforations 24 is not particularly limited, but may be, for example, 0.5 to 1.5 mm. The number of perforations 24 is not particularly limited, and may be one or two or more. For example, a plurality of perforations 24 may be provided on the circumference of the cooling section 20B.

[0073] The cooling section 20B may be rod-shaped with an axial length of, for example, 7 to 28 mm. For example, the axial length of the cooling section 20B may be 18 mm. The cooling section 20B may have a substantially circular axial cross section with a diameter of 5 to 10 mm. For example, the diameter of the cooling section may be approximately 7 mm.

[0074] (3) Filter part 20C The configuration of the filter part 20C is not particularly limited, and may be composed of one or more packed layers. The outside of the packed layer may be wrapped with one or more sheets of wrapping paper. The airflow resistance of the filter part 20C can be appropriately changed depending on the amount, material, etc. of the packing filled in the filter part 20C. For example, when the packing is cellulose acetate fiber, the airflow resistance can be increased by increasing the amount of cellulose acetate fiber filled in the filter part 20C. When the packing is cellulose acetate fiber, the packing density of the cellulose acetate fiber is 0.13 to 0.18 g / cm. 3 The airflow resistance is a value measured using an airflow resistance measuring device (product name: SODIMAX, manufactured by SODIM).

[0075] The circumferential length of the filter portion 20C is not particularly limited, but is preferably 16 to 25 mm, more preferably 20 to 24 mm, and even more preferably 21 to 23 mm. The axial length of the filter portion 20C (the horizontal direction in FIG. 3) can be selected from 4 to 10 mm, and is selected so that the airflow resistance is 15 to 60 mmH2O / seg. The axial length of the filter portion 20C is preferably 5 to 9 mm, more preferably 6 to 8 mm. The cross-sectional shape of the filter portion 20C is not particularly limited, but may be, for example, circular, elliptical, polygonal, etc. In addition, a flavor-containing breakable capsule, flavor beads, or flavor may be directly added to the filter portion 20C.

[0076] The filter portion 20C may have a center hole portion as the first segment 25. The center hole portion is composed of a first packed layer 25a having one or more hollow portions and an inner plug wrapper (inner wrapping paper) 25b that covers the packed layer. The center hole portion functions to increase the strength of the mouthpiece portion. The center hole portion may not have an inner plug wrapper 25b and its shape may be maintained by thermoforming. The first packed layer 25a may be, for example, a rod with an inner diameter of 5.0 to 1.0 mm, densely packed with cellulose acetate fibers, to which a plasticizer containing triacetin is added in an amount of 6 to 20% by mass relative to the mass of cellulose acetate and hardened. Because the first packed layer 25a has a high fiber packing density, during inhalation, air and aerosol flow only through the hollow portion and hardly flow within the first packed layer 25a. Because the first packed layer 25a in the center hole portion is a fiber packed layer, the external feel during use is less likely to cause discomfort to the user. The filter portion 20C may also have a second segment 26. The second segment 26 is composed of a second packed layer 26a and an inner plug wrapper (inner wrapping paper) 26b that covers the packed layer.

[0077] The first segment 25 and the second segment 26 are connected by an outer plug wrapper (outer wrapping paper) 27. The outer plug wrapper 27 can be, for example, a cylindrical piece of paper. The tobacco rod portion 20A, the cooling portion 20B, and the connected first segment 25 and second segment 26 are connected by a mouthpiece lining paper 28. These connections can be made, for example, by applying glue such as vinyl acetate glue to the inner surface of the mouthpiece lining paper 28 and wrapping the three components around it. These components may also be connected in multiple layers using multiple lining papers.

[0078] (Non-combustion heating type flavor inhalation system) An example of a non-combustion heating type flavor inhalation system according to this embodiment is shown in Fig. 4. In Fig. 4, the non-combustion heating type flavor inhalation system includes a non-combustion heating type flavor inhaler 20 and a heating device 10 that heats the tobacco rod portion 20A from the outside.

[0079] The heating device 10 includes a body 11, a heater 12, a metal tube 13, a battery unit 14, and a control unit 15. The body 11 has a cylindrical recess 16, and the heater 12 and metal tube 13 are disposed in a position corresponding to the tobacco rod portion 20A to be inserted therein. The heater 13 may be an electric resistance heater, and is heated by power supplied from the battery unit 14 in response to instructions from a temperature-controlling control unit 15. The heat generated by the heater 12 is transferred to the tobacco rod portion 20A through the metal tube 13, which has high thermal conductivity. While FIG. 4 shows an embodiment in which the heating device 10 heats the tobacco rod portion 20A from the outside, it may also heat from the inside. The heating temperature of the heating device 10 is not particularly limited, but is preferably 400°C or less, more preferably 150 to 400°C, and even more preferably 200 to 350°C. The heating temperature refers to the temperature of the heater 12 of the heating device 10.

[0080] [Non-combustion, non-heating type flavor inhaler and its manufacturing method] A method for manufacturing a non-combustion, non-heating flavor inhaler according to this embodiment includes the steps of isolating and purifying neophytadiene from tobacco leaves to obtain a neophytadiene composition by the method according to this embodiment, adding the neophytadiene composition to a substrate, and manufacturing a non-combustion, non-heating flavor inhaler including the substrate. Furthermore, the non-combustion, non-heating flavor inhaler according to this embodiment includes the neophytadiene composition according to this embodiment. The method for manufacturing a non-combustion, non-heating flavor inhaler according to this embodiment obtains the neophytadiene composition by the method according to this embodiment, thereby efficiently isolating and purifying neophytadiene from tobacco leaves and efficiently manufacturing a non-combustion, non-heating flavor inhaler that can provide a good flavor. Furthermore, the non-combustion, non-heating flavor inhaler according to this embodiment can provide a good flavor because it includes the neophytadiene composition obtained by the method according to this embodiment.

[0081] In the method for producing a non-combustion and non-heating type flavor inhaler according to this embodiment, the method for adding the neophytadiene composition to the base material can be the same as in the method for producing the non-combustion and non-heating type flavor inhaler described above.

[0082] FIG. 5 shows one embodiment of a non-combustion, non-heating flavor inhaler according to the present embodiment. The non-combustion, non-heating flavor inhaler 30 includes a power supply unit 30D, a cartridge 30E, and a tobacco capsule 30F. The non-combustion, non-heating flavor inhaler 30 has a shape extending from a non-suction end u (upstream) to a mouth end d (downstream). The cartridge 30E is detachable from the power supply unit 30D. The tobacco capsule 30F is detachable from the cartridge 30E.

[0083] (1) Tobacco capsules FIG. 6 shows an example of a tobacco capsule 30F. As shown in FIG. 6, the tobacco capsule 30F is a tobacco rod portion and contains a flavor source 300 therein. The flavor source 300 includes a tobacco material containing the neophytadiene composition according to this embodiment. The tobacco capsule 30F is connected to a cartridge 30E. Specifically, a portion of the tobacco capsule 30F is housed within the cartridge 30E.

[0084] The tobacco capsule 30F has a housing 310 that houses the flavor source 300, a mesh body 320, a nonwoven fabric 330, and a cap 340. Aerosol atomized by the atomization unit 220 (described later) is introduced into the housing 310 through the mesh body 320, and is imparted with flavor by coming into contact with the flavor source 300. The aerosol is then inhaled by the user through the nonwoven fabric 330. In this way, the non-combustion, non-heating type flavor inhaler 30 can impart flavor to the aerosol without heating the flavor source 300. Furthermore, substantially no aerosol is generated from the flavor source 300.

[0085] In the direction of aerosol flow, the length of the tobacco capsule 30F (container 310) is preferably 40 mm or less, more preferably 25 mm or less. Furthermore, in the direction of aerosol flow, the length is preferably 1 mm or more, more preferably 5 mm or more. In a direction perpendicular to the direction of aerosol flow, the maximum length of the container 310 of the tobacco capsule 30F (container 310) is preferably 20 mm or less, more preferably 10 mm or less. Furthermore, in a direction perpendicular to the direction of aerosol flow, the maximum length of the tobacco capsule 30F (container 310) is preferably 1 mm or more, more preferably 3 mm or more.

[0086] The tobacco-containing flavor source 300 is composed of raw material pieces that impart flavor to the aerosol. The lower limit of the raw material piece size is preferably 0.2 to 1.2 mm, more preferably 0.2 to 0.7 mm. The smaller the size of the raw material pieces constituting the flavor source 300, the greater the specific surface area, thereby facilitating the release of flavor components. Examples of raw material pieces constituting the flavor source 300 include shredded tobacco containing the neophytadiene composition of the present embodiment, or a molded product obtained by molding a tobacco material containing the neophytadiene composition of the present embodiment into a granular form. The flavor source 300 may contain flavorings derived from plants other than tobacco (e.g., mint, herbs, etc.) or menthol. Furthermore, the tobacco-containing flavor source 300 may contain a flavoring agent. Examples of flavoring agents include materials that exhibit sweetness, sourness, saltiness, umami, bitterness, astringency, richness, spiciness, harshness, and astringency. Examples of sweetening materials include sugars, sugar alcohols, sweeteners, etc. Examples of sugars include monosaccharides, disaccharides, oligosaccharides, polysaccharides, etc. Examples of sweeteners include natural sweeteners, synthetic sweeteners, etc.

[0087] The raw material pieces are obtained, for example, by sieving in accordance with JIS Z 8815 using a stainless steel sieve conforming to JIS Z 8801. For example, the raw material pieces are sieved using a stainless steel sieve with 0.71 mm openings for 20 minutes by dry mechanical shaking to obtain raw material pieces that pass through the stainless steel sieve with 0.71 mm openings. Next, the raw material pieces are sieved using a stainless steel sieve with 0.212 mm openings for 20 minutes by dry mechanical shaking to remove the raw material pieces that pass through the stainless steel sieve with 0.212 mm openings. In other words, the raw material pieces that make up the flavor source 300 are raw material pieces that pass through the stainless steel sieve (mesh opening = 0.71 mm) that defines the upper limit but do not pass through the stainless steel sieve (mesh opening = 0.212 mm) that defines the lower limit. Therefore, the lower limit of the size of the raw material pieces constituting the flavor source 300 is defined by the opening size of the stainless steel sieve that defines the lower limit, and the upper limit of the size of the raw material pieces constituting the flavor source 300 is defined by the opening size of the stainless steel sieve that defines the upper limit.

[0088] From the viewpoint of providing a good flavor, the amount of flavor source 300 contained in the container 310 is preferably 300 mg or more, and more preferably 350 mg or more.

[0089] (2) Power supply unit An example of a power supply unit 30D is shown in FIG. 7. The power supply unit 30D has a battery 110. The battery 110 may be a disposable battery or a rechargeable battery. The initial output voltage of the battery 110 is preferably in the range of 1.2 V to 4.2 V. The battery capacity of the battery 110 is preferably in the range of 100 mAh to 1000 mAh.

[0090] (3) Cartridge An example of a cartridge 30E is shown in Figures 8 and 9. Figure 8 is a cross-sectional view of an example of cartridge 30E, and Figure 9 is a diagram showing its internal structure. Cartridge 30E has a reservoir 210, an atomizing section 220, a flow path forming body 230, an outer frame body 240, and an end cap 250. Cartridge 30E has a first flow path 200X, which serves as an aerosol flow path, and is located downstream of atomizing section 220.

[0091] The reservoir 210 stores the aerosol source 200. The reservoir 210 is located around the flow path forming body 230 in a cross section perpendicular to the aerosol flow direction (direction from the non-suction end to the suction end (upstream to downstream)). The reservoir 210 is located in the gap between the flow path forming body 230 and the outer frame body 240. The reservoir 210 is formed, for example, from a porous body such as a resin web or cotton. The reservoir 210 may also be formed from a tank that stores the liquid aerosol source 200. Examples of the aerosol source 200 include glycerin and propylene glycol.

[0092] The atomization unit 220 atomizes the aerosol source 200 using power supplied from the battery 110 without combustion. The atomization unit 220 is composed of a heating wire (coil) wound at a predetermined pitch. The atomization unit 220 is preferably composed of a heating wire having a resistance value in the range of 1.0 to 3.0 Ω. The predetermined pitch is equal to or greater than the value at which the heating wires do not touch each other, and is preferably smaller than that. The predetermined pitch is preferably, for example, 0.40 mm or less. The predetermined pitch is preferably constant to stabilize the atomization of the aerosol source 200. The predetermined pitch is the distance between the centers of adjacent heating wires.

[0093] The flow path forming body 230 has a cylindrical shape that forms a first flow path 200X extending along the aerosol flow direction. The outer frame body 240 has a cylindrical shape that houses the flow path forming body 230. The outer frame body 240 extends downstream of the end cap 250 and houses a part of the tobacco capsule 30F. The end cap 250 is a cap that closes the gap between the flow path forming body 230 and the outer frame body 240 from the downstream side. The end cap 250 prevents the aerosol source 200 stored in the reservoir 210 from leaking toward the tobacco capsule 30F.

[0094] [Combustion-type flavor inhaler and its manufacturing method] The method for manufacturing a combustion-type flavor inhaler according to the present embodiment includes the steps of isolating and purifying neophytadiene from tobacco leaves to obtain a neophytadiene composition by the method according to the present embodiment, adding the neophytadiene composition to a substrate, and manufacturing a combustion-type flavor inhaler including the substrate. The combustion-type flavor inhaler according to the present embodiment also includes the neophytadiene composition according to the present embodiment. The method for manufacturing a combustion-type flavor inhaler according to the present embodiment obtains the neophytadiene composition by the method according to the present embodiment, thereby efficiently isolating and purifying neophytadiene from tobacco leaves and efficiently manufacturing a combustion-type flavor inhaler that can provide a good flavor. The combustion-type flavor inhaler according to the present embodiment also provides a good flavor because it includes the neophytadiene composition obtained by the method according to the present embodiment.

[0095] In the method for producing a combustion-type flavor inhaler according to this embodiment, the method for adding the neophytadiene composition to the base material can be the same as in the method for producing a non-combustion-heating flavor inhaler described above.

[0096] FIG. 10 shows an example of a combustion-type flavor inhaler according to the present embodiment. As shown in FIG. 10, the combustion-type flavor inhaler 400 includes a tobacco-containing segment 401 and a filter segment 402 adjacent to the tobacco-containing segment 401. The tobacco-containing segment 401 includes a tobacco filler 403 containing a tobacco material containing the neophytadiene composition according to the present embodiment, and cigarette paper 404 wrapped around the tobacco filler 403. As described above, the cigarette paper 404 may contain the neophytadiene composition according to the present embodiment. The filter segment 402 is not particularly limited as long as it functions as a general filter. For example, synthetic fiber tow (also simply referred to as "tow") or a cylindrical material such as paper can be used. The tobacco-containing segment 401 and the filter segment 402 are connected by a tipping paper member 405 wrapped around the tobacco-containing segment 401 and the filter segment 402. The tipping paper member 405 may have ventilation holes in part of its periphery. The number of ventilation holes may be one or more, for example, 10 to 40. When there are multiple ventilation holes, the ventilation holes may be arranged, for example, in a ring shape in a line around the outer periphery of the tipping paper member 405. The multiple ventilation holes may be arranged at approximately regular intervals. By providing ventilation holes, air is taken into the filter segment 402 through the ventilation holes during inhalation. By diluting the mainstream smoke with outside air from the ventilation holes, a product with a desired tar value can be designed.

[0097] The user can enjoy the flavor of tobacco by lighting the tip of tobacco-containing segment 401 and drawing on the mouth end of filter segment 402. The number of filter segment 402 is not limited to one, and multiple filter segments with different functions may be connected together. [Example]

[0098] Specific examples of this embodiment will be described below, but the present invention is not limited to these.

[0099] [Reference Example 1](Examination of Tobacco Varieties) Weighed 200 g of fine powder of yellow or Burley tobacco leaves and put it into a 2500 ml sealed stainless steel container. Next, 2000 ml of hexane (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd., for high performance liquid chromatography) was added, and then extracted for 3 hours with stirring in a sealed state in a warm bath at 40°C. After extraction, the hexane solution and the extraction residue were separated using a stainless steel mesh with a mesh opening of 250 μm, and about 1500 ml of hexane solution was obtained. Prepared a 0.1 mass% sulfuric acid aqueous solution in advance, and mixed the 0.1 mass% sulfuric acid aqueous solution and the hexane solution obtained above at a solution ratio (volume ratio) of sulfuric acid aqueous solution:hexane solution = 1:3. Further, 200 g of sodium chloride was added and shaken well in a separatory funnel to perform a liquid-liquid extraction operation. At this time, components represented by nicotine migrated to the lower layer sulfuric acid aqueous solution, and hydrophobic active components of tobacco leaves migrated to the upper layer organic layer. After sufficient standing, the organic layer was taken out, 200 g of anhydrous sodium sulfate was added and stirred to perform a dehydration operation in the organic layer. Insoluble matters were removed by filtration through filter paper. Further, hexane was removed under reduced pressure using a rotary evaporator (manufactured by Buchi, Japan), and 5.16 g (yield 2.58%) of dry matter was obtained from the yellow variety and 5.4 g (yield 2.7%) of dry matter was obtained from the Burley variety. Each dry matter was dissolved in a hexane solvent, and the obtained solution was analyzed by GC / MS under the conditions shown below.

[0100] Gas Chromatography-Mass Spectrometry (GC / MS) with Mass Spectrometer Apparatus: 7890A / 5975C GC / MSD manufactured by Agilent Technologies <GC Conditions> Column: HP-5MS (manufactured by Agilent Technologies) Inner diameter 0.25 mm × length 30 m, film thickness 0.25 μm Injection volume: 1 μl Injection mode: Split (10:1) Inlet temperature: 270°C Septum purge flow rate: 5 ml / min Carrier gas: Helium (He) Column flow rate: 1 ml / min (constant flow mode) Oven temperature: 40°C (3 minutes) - 4°C / min - 280°C (20 minutes) Transfer line temperature: 280 °C Solvent waiting time: 4 minutes Ionization method: Electron impact ionization method (EI method), 70 eV Ion source temperature: 230 °C Quadrupole temperature: 150 °C Measurement mode: Scan MS scan range: m / z 26 - 450 Threshold: 50 Sampling rate: 2

[0101] From the total ion chromatogram obtained by the above analysis using GC / MS, the single ion (m / z = 68) included in the MS pattern of neophytadiene was extracted, and the peak area values were compared. As a result, it was confirmed that the peak area value of neophytadiene in the yellow variety was about 1.4 times larger than that in the burley variety. From this, it was found that both the yellow variety and the burley variety are useful as tobacco leaves, but the yellow variety is more preferable from the viewpoint of higher neophytadiene yield.

[0102] [Reference Example 2] (Examination of extraction solvent) Weighed 5 g of tobacco leaf fine powder into a screw tube of a 100 ml container, added 50 ml of an organic solvent (n-hexane, ethyl acetate, ethanol, or methanol), mixed well, and then left to stand at room temperature for one day and night. A small amount of anhydrous sodium sulfate was added to the extract obtained by filtering the mixed solution with filter paper for dehydration, followed by filtration again with filter paper, and the organic solvent was removed under reduced pressure. The dried solid was dissolved in the same solvent as the solvent used for extraction so that the concentration of the obtained dried solid became 4% by mass, and the solution was analyzed by GC / MS under the conditions shown in Reference Example 1. The total ion chromatograms obtained by the analysis of the n-hexane, ethyl acetate, ethanol, and methanol solutions by GC / MS are shown in FIGS. 11 to 14.

[0103] 11 to 14, it was found that neophytadiene (retention time 38.9 minutes) was extracted from the fine leaf tobacco powder when extracted with each solvent. Furthermore, it was confirmed that peaks of components (retention time 38 minutes or less) that are not necessary for the purification of neophytadiene were present in the extractions using ethyl acetate, ethanol, and methanol. This indicates that, of the four solvents, hexane, which contains as few peaks of these unnecessary components as possible, is the most suitable extraction solvent.

[0104] [Example 1] (Process A1) 200 g of flue-cured tobacco powder was weighed and placed in a 2500 ml sealed stainless steel container. 2000 ml of hexane (Fujifilm Wako Pure Chemical Industries, high-performance liquid chromatograph grade) was then added, and the mixture was extracted for 3 hours in a sealed, stirring bath at 40°C. After extraction, the hexane solution and extraction residue were separated using a 250 μm stainless steel mesh, yielding approximately 1500 ml of hexane solution.

[0105] (Process B1) A 0.1% by weight aqueous sulfuric acid solution was prepared in advance. This 0.1% by weight aqueous sulfuric acid solution was mixed with the previously prepared hexane solution at a volume ratio of 1:3 (sulfuric acid aqueous solution:hexane solution) to obtain 500 ml of a mixed solution. 200 g of salt was then added, and the mixture was thoroughly shaken in a separatory funnel to perform liquid-liquid extraction. During this process, components such as nicotine migrated to the lower sulfuric acid aqueous solution, while the hydrophobic active ingredients of tobacco leaf migrated to the upper organic layer. After allowing to stand for a sufficient period, the organic layer was removed, and approximately 200 g of anhydrous sodium sulfate was added and stirred to dehydrate the organic layer. Insoluble matter was removed by paper filtration. The resulting solution was analyzed by GC / MS under the conditions described in Reference Example 1. The results are shown in Figure 15. The chromatogram shown in Figure 15 confirmed the presence of neophytadiene (retention time 38.9 minutes) and the removal of alkaloids such as nicotine (retention time 24.6 minutes) by the liquid-liquid extraction procedure.

[0106] (Process C1) Methanol was added to the organic layer (hexane layer) obtained above to a volume ratio of 2:1 (organic layer:methanol), mixed thoroughly, and then left to stand overnight in a refrigerator (4°C) for liquid-liquid extraction. During this process, CBT and other tobacco leaf surface resin components migrated to the lower methanol solution, while neophytadiene and higher hydrocarbons migrated to the upper hexane solution. The hexane and methanol solutions obtained by this extraction were analyzed by GC / MS under the conditions described in Reference Example 1. The results for the hexane solution are shown in Figure 16, and the results for the methanol solution are shown in Figure 17. The chromatograms shown in Figures 16 and 17 confirm that CBT was selectively removed by this process.

[0107] (Steps D1, E1 and F1) Hexane was removed from the hexane solution obtained above under reduced pressure using a rotary evaporator (manufactured by Nippon Buchi Co., Ltd.), yielding 3.15 g of a dry product (yield: 1.58%). 300 mL of methanol was added to the dry product, and the dry product was dispersed and dissolved in the methanol while being subjected to ultrasonic waves. The resulting solution was allowed to stand in a refrigerator (4°C) for 1 hour, after which the supernatant, from which the precipitate had been removed, was recovered. The supernatant was analyzed by GC / MS under the conditions described in Reference Example 1. The results are shown in Figure 18. It was confirmed that higher hydrocarbons were selectively removed by this process.

[0108] (Steps G1 and H1) Methanol was removed from the supernatant liquid obtained above under reduced pressure using a rotary evaporator (manufactured by Nippon Buchi). Hexane (15 mL) was added to the resulting dried product (1.49 g) to dissolve it. Neophytadiene was isolated and purified from the resulting solution using normal-phase HPLC. The HPLC used was a 1260 Infinity (trade name, manufactured by Agilent Technologies). The HPLC separation conditions were as follows: Column: YMC-Pack SIL / S-5μm / 12nm ·Mobile phase: Hex(2min)→Hex:AcOEt(95:5)(1min)→Hex(12min) ·Flow rate: 7mL / min Detector: Diode array detector (220 nm) Sample injection volume: 500 μL

[0109] After separation under the above conditions, the signal detected at 220 nm was fractionated in a fraction collector. The fractionated fractions are shown within the dashed line in Figure 19. The fractions were collected, and the resulting sample was concentrated and dried to obtain a colorless, transparent neophytadiene composition. The total ion chromatogram obtained by GC / MS analysis showed a neophytadiene yield of 65.0%, confirming that neophytadiene can be isolated and purified from tobacco leaves in high yield. The resulting neophytadiene composition was dissolved in methanol and analyzed by GC / MS under the conditions shown in Reference Example 1. The results are shown in Figure 20. The purity of the neophytadiene composition was calculated to be 99.83%, confirming that the method of this embodiment can isolate and purify neophytadiene with high purity.

[0110] [Example 2] The neophytadiene composition (alcohol preparation) obtained in Example 1 was added to tobacco shreds at 1000 ppm by mass. The resulting aromatized shredded sheets were dried and packed into the tobacco rod portion of an externally heated, non-combustion heating type flavor inhaler for evaluation. Unaromatized shredded sheets were also packed into the tobacco rod portion of an externally heated, non-combustion heating type flavor inhaler for evaluation. As a result, the aromatized shredded sheets exhibited the essential tobacco aroma better than the unaromatized shredded sheets.

[0111] [Example 3] The neophytadiene composition (alcohol preparation) obtained in Example 1 was added to tobacco shreds at 500 ppm by mass. The obtained flavored tobacco shreds were dried, packed into the tobacco-containing segment of a combustion-type flavor inhaler, and evaluated. Unflavored tobacco shreds were also packed into the tobacco-containing segment of a combustion-type flavor inhaler and evaluated in the same manner. As a result, the flavored tobacco shreds expressed the essential inherent tobacco aroma better than the unflavored tobacco shreds. [Explanation of symbols]

[0112] 10 Heating device 11 Body 12 Heater 13 Metal tube 14 Battery unit 15 Control Unit 16 Recess 17 Ventilation holes 20 Non-combustion heating type flavor inhaler 20A Tobacco rod part 20B Cooling section 20C filter section 21 Tobacco filler 22 Rolling Paper 23 Paper tube 24 perforation 25 First Segment 25a 1st packed bed 25b inner plug wrapper 26 Second Segment 26a 2nd packed bed 26b Inner plug wrapper 27 Outer plug wrapper 28 Lining Paper 30 Non-burning, non-heating flavor inhaler 30D Power Supply Unit 30E Cartridge 30F Tobacco Capsule u Non-suction end d Mouth end 110 Batteries 200 Aerosol Sources 210 Reservoir 220 Atomization section 230 Flow path formation body 240 Outer frame 240 250 end cap 200X First flow path 300 Flavor source 310 Containment Unit 320 mesh body 330 Nonwoven fabric 340 Cap 400 Combustion-type Flavor Inhaler 401 Tobacco-containing segment 402 Filter Segment 403 Tobacco fillers 404 Rolling Paper 405 Chip paper material

Claims

1. A method for isolating and purifying neophytadiene from tobacco leaves, comprising the steps of: a step of adding an aprotic solvent to tobacco leaves to perform an extraction operation, and removing the extraction residue to obtain an extract (A1); a step of adding an acidic aqueous solution to the extract (A1) to perform liquid-liquid extraction, and removing the aqueous layer to obtain an extract (B1); a step of adding a protic solvent to the extract (B1) to perform liquid-liquid extraction, and removing the protic solvent layer to obtain an extract (C1); a step of removing the solvent from the extract (C1) to obtain a dry product (D1); a step of adding a protic solvent to the dried product (D1) to dissolve the dried product (D1) and obtain a solution (E1); removing a precipitate formed in the solution (E1) to obtain a supernatant (F1); a step of replacing the solvent of the supernatant (F1) with an aprotic solvent to obtain a solution (G1); isolating and purifying neophytadiene from the solution (G1) by normal phase high performance liquid chromatography; A method comprising:

2. A method for isolating and purifying neophytadiene from tobacco leaves, comprising the steps of: a step of adding an aprotic solvent to tobacco leaves to perform an extraction operation, and removing the extraction residue to obtain an extract (A2); a step of adding an acidic aqueous solution to the extract (A2) to perform liquid-liquid extraction, and removing the aqueous layer to obtain an extract (B2); a step of removing the solvent from the extract (B2) to obtain a dry product (C2); a step of adding a protic solvent to the dried product (C2) to dissolve the dried product (C2) and obtain a solution (D2); a step of removing a precipitate formed in the solution (D2) to obtain a supernatant (E2); a step of adding an aprotic solvent to the supernatant (E2) to perform liquid-liquid extraction, and removing the protic solvent layer to obtain a solution (F2); isolating and purifying neophytadiene from the solution (F2) by normal phase high performance liquid chromatography; A method comprising:

3. 3. The method according to claim 1, wherein the acidic aqueous solution has a pH of 3 or less.

4. 3. The method according to claim 1, wherein the aprotic solvent is at least one solvent selected from the group consisting of hexane, heptane, and ethyl acetate.

5. 3. The method of claim 1, wherein the protic solvent is methanol.

6. 3. The method of claim 1 or 2, wherein the tobacco leaf is at least one of flue-cured and burley.

7. 2. The method according to claim 1, wherein the step of obtaining the solution (G1) is a step of adding an aprotic solvent to the supernatant (F1) to perform liquid-liquid extraction, and removing a protic solvent layer to obtain the solution (G1).

8. 2. The method according to claim 1, wherein the step of obtaining the solution (G1) is a step of removing the solvent from the supernatant (F1) and then adding an aprotic solvent to obtain the solution (G1).

9. 3. The method according to claim 1, wherein the neophytadiene composition obtained by the isolation and purification contains neophytadiene in an amount of 95% by mass or more.

10. A step of isolating and purifying neophytadiene from tobacco leaves by the method according to claim 1 or 2 to obtain a neophytadiene composition; adding the neophytadiene composition to a substrate; a step of manufacturing a non-combustion heating type flavor inhaler including the base material; A method for manufacturing a non-combustion heating type flavor inhaler, comprising:

11. A step of isolating and purifying neophytadiene from tobacco leaves by the method according to claim 1 or 2 to obtain a neophytadiene composition; adding the neophytadiene composition to a substrate; a step of manufacturing a non-combustion, non-heating flavor inhaler comprising the base material; A method for manufacturing a non-burning, non-heating flavor inhaler, comprising:

12. A step of isolating and purifying neophytadiene from tobacco leaves by the method according to claim 1 or 2 to obtain a neophytadiene composition; adding the neophytadiene composition to a substrate; manufacturing a combustion-type flavor inhaler comprising the substrate; A method for manufacturing a combustion type flavor inhaler, comprising:

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