Method for separating and purifying neophytadiene from tobacco leaf, neophytadiene composition, non-combustion heating-type flavor suction device, non-combustion non-heating-type flavor suction device, combustion-type flavor suction device, and method for producing same

Abstract

Provided is a method whereby neophytadiene can be separated and purified from tobacco leaves with high yield and high purity. The method comprises: a step in which an aprotic solvent is added to tobacco leaves to perform an extraction operation, and extraction residues are removed to obtain an extraction solution (A1); a step for adding an acidic aqueous solution to the extract (A1) and performing liquid-liquid extraction to remove the water layer to obtain an extract (B1); a step in which a protic solvent is added to the extract (B1) and liquid-liquid extraction is carried out, and a protic solvent layer is removed to obtain an extract (C1); a step for obtaining a dried solid (D1) by removing the solvent from the extract (C1); a step for obtaining a solution (E1) by adding a protic solvent to the dried solid (D1); a step for obtaining a supernatant (F1) by removing a precipitate generated in the solution (E1); a step for obtaining a solution (G1) by replacing the solvent of the supernatant (F1) with an aprotic solvent; and a step for separating and purifying neophytadiene from the solution (G1) by normal phase HPLC.

Description

Technical Field

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

[0002] Neophytadiene is a terpenoid compound contained in tobacco leaves, which is related to the characteristic aroma of tobacco plants and is therefore attracting attention as an important component that reflects the aroma quality. Essential oils containing neophytadiene are known to have anti-inflammatory effects (Non-Patent Document 1), but in recent years, it has been clarified that neophytadiene monomers exert anti-inflammatory effects, and therefore have attracted attention (Non-Patent Document 2).

[0003] As a substance containing neophytadiene, in plants, in addition to tobacco, algae are also known (non-patent document 2). It is reported that neophytadiene is derived from phytol released by hydrolysis of a part of chlorophyll a and b contained in plants in the lumen, and it is also reported that neophytadiene is detected in milk and beef of cattle fed with grass containing neophytadiene (non-patent document 3).

[0004] As a method for obtaining neophytadiene monomers, a synthesis reaction is mainly used. In the synthesis reaction, chlorophyll is first hydrolyzed under alkaline conditions to generate phytol. Concentrated sulfuric acid is added to the generated phytol and heated at 170°C to cause an elimination reaction to occur, thereby generating neophytadiene (Patent Document 1).

[0005] On the other hand, when isolating neophytadiene from a mixture of plants, etc., it is often purified into essential oils containing other substances, and there are few reports on the separation and purification of neophytadiene monomers. For example, in non-patent literature 4, a method is described: as a process for separating and purifying neophytadiene from tobacco, after extracting yellow tobacco with a hexane solvent, eluting the hexane layer using a silica column chromatography, replacing the solvent with acetone, crystallizing and removing saturated hydrocarbons, and eluting them using alumina chromatography.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: Japanese Patent Application No. 2012-529893

[0009] Non-patent literature:

[0010] Non-patent literature 1: Adeosun TE, Ogunwande IA, Avoseh ON, Raji IP, LawalOA. Composition and Anti-inflammatory Activity of Essential Oil of Jatrop hacurcas. Nat Prod Commun. 2017Mar; 12(3): 439-440. PMID: 30549905.

[0011] Non-patent literature 2: Bhardwaj, M., Sali, VK, Mani, S. et al. Neophytadiene from Turbinaria ornate Suppresses LPS-Induced Inf lammatory Response in RAW264.7 Macrophages and Sprague Dawley Rats. Inflammation 43,937-950 (2020)

[0012] Non-patent document 3: Research on the establishment of traceability based on the volatile component characteristics of milk produced from pasture-raising and the volatile components in the milk ("Volatile Components of Pasture-Raised Milk" Research on the Establishment of Volatile Ingredients in Special Milk Milk"), Yasuko Ueda, Hokkaido University, PhD (Agriculture) B No. 6987 (2016)

[0013] Non-patent document 4: Flue-cured Tobacco.II. Neophytadiene Summary of the invention

[0014] Problems to be solved by the invention

[0015] However, the method described in Non-Patent Document 4 cannot separate and purify neophytadiene with high yield and high purity. From the perspective of expanding the effective utilization of neophytadiene, it is desired to develop a method that can separate and purify neophytadiene from tobacco leaves with high yield and high purity.

[0016] The object of the present invention is to provide a method for separating and purifying neophytadiene from tobacco leaves with high yield and high purity, to provide a neophytadiene composition obtained by the method, to provide a non-combustion heating type flavor inhaler containing the neophytadiene composition, a non-combustion non-heating type flavor inhaler, and a combustion type flavor inhaler.

[0017] Solutions to the problem

[0018] The present invention includes the following embodiments.

[0019] [1] A method for separating and purifying neophytadiene from tobacco leaves, the method comprising:

[0020] A step of adding an aprotic solvent to the tobacco leaves for extraction, removing the extraction residue, and obtaining an extract (A1);

[0021] A step of adding an acidic aqueous solution to the extract (A1) for liquid-liquid extraction, removing the aqueous layer, and obtaining an extract (B1);

[0022] 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);

[0023] A step of removing the solvent from the extract (C1) to obtain a dry solid (D1);

[0024] A step of adding a protic solvent to the dry solid (D1) to dissolve the dry solid (D1) to obtain a solution (E1);

[0025] A step of removing the precipitate generated in the solution (E1) to obtain a supernatant (F1);

[0026] A step of replacing the solvent of the supernatant (F1) with an aprotic solvent to obtain a solution (G1); and

[0027] A step of separating and purifying neophytadiene from the solution (G1) by normal phase high performance liquid chromatography.

[0028] [2] A method for separating and purifying neophytadiene from tobacco leaves, the method comprising:

[0029] A step of adding an aprotic solvent to the tobacco leaves for extraction, removing the extraction residue, and obtaining an extract (A2);

[0030] A step of adding an acidic aqueous solution to the extract (A2) for liquid-liquid extraction, removing the aqueous layer, and obtaining an extract (B2);

[0031] A step of removing the solvent from the extract (B2) to obtain a dry solid (C2);

[0032] A step of adding a protic solvent to the dry solid (C2) to dissolve the dry solid (C2) to obtain a solution (D2);

[0033] A step of removing the precipitate generated in the solution (D2) to obtain a supernatant (E2);

[0034] A step of adding an aprotic solvent to the supernatant (E2) to perform liquid-liquid extraction, removing the protic solvent layer, and obtaining a solution (F2); and

[0035] A step of separating and purifying neophytadiene from the solution (F2) by normal phase high performance liquid chromatography.

[0036] [3] The method according to [1] or [2], wherein:

[0037] The pH of the acidic aqueous solution is 3 or less.

[0038] [4] The method according to any one of [1] to [3], wherein

[0039] The aprotic solvent is at least one solvent selected from hexane, heptane and ethyl acetate.

[0040] [5] The method according to any one of [1] to [4], wherein

[0041] The protic solvent is methanol.

[0042] [6] The method according to any one of [1] to [5], wherein

[0043] The tobacco leaf is at least one of yellow tobacco and burley tobacco.

[0044] [7] The method according to [1], wherein:

[0045] 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).

[0046] [8] The method according to [1], wherein:

[0047] The step of obtaining the solution (G1) is a step of removing the solvent of the supernatant (F1) and then adding an aprotic solvent to obtain the solution (G1).

[0048] [9] The method according to any one of [1] to [8], wherein

[0049] The neophytadiene composition obtained by separation and purification contains more than 95% by mass of neophytadiene.

[0050]

[10] A method for manufacturing a non-combustion heating type fragrance inhaler, comprising:

[0051] A step of separating and purifying neophytadiene from tobacco leaves by the method described in any one of [1] to [9] to obtain a neophytadiene composition;

[0052] The step of adding the neophytadiene composition to a substrate; and

[0053] A process for manufacturing a non-combustion heating type flavor inhaler including the substrate.

[0054]

[11] A method for manufacturing a non-combustion and non-heating type fragrance inhaler, comprising:

[0055] A step of separating and purifying neophytadiene from tobacco leaves by the method according to any one of claims [1] to [9] to obtain a neophytadiene composition;

[0056] The step of adding the neophytadiene composition to a substrate; and

[0057] The invention relates to a process for manufacturing a non-combustion and non-heating type flavor inhaler including the substrate.

[0058]

[12] A method for manufacturing a combustion type aroma inhaler, comprising:

[0059] A step of separating and purifying neophytadiene from tobacco leaves by the method described in any one of [1] to [9] to obtain a neophytadiene composition;

[0060] The step of adding the neophytadiene composition to a substrate; and

[0061] A process for manufacturing a combustion-type flavor inhaler including the substrate.

[0062]

[13] A neophytadiene composition comprising 95% by mass or more of neophytadiene, the composition being obtained by separating and purifying neophytadiene from tobacco leaves by the method described in any one of [1] to [9].

[0063]

[14] A non-combustion heating type flavor inhaler comprising the neophytadiene composition described in

[13] .

[0064]

[15] A non-combustion and non-heating flavor inhaler comprising the neophytadiene composition described in

[13] .

[0065]

[16] A combustion-type flavor inhaler comprising the neophytadiene composition described in

[13] .

[0066] Effects of the Invention

[0067] According to the present invention, a method for separating and purifying neophytadiene from tobacco leaves with high yield and high purity, a neophytadiene composition obtained by the method, a non-combustion and heating type flavor inhaler containing the neophytadiene composition, a non-combustion and non-heating type flavor inhaler, and a combustion type flavor inhaler can be provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0068] Figure 1 This is a flowchart showing an example of a method according to the first embodiment of the present invention.

[0069] Figure 2 This is a flowchart showing an example of a method according to the second embodiment of the present invention.

[0070] Figure 3 Schematic diagram showing an example of the non-combustion heating type flavor inhaler according to the present embodiment.

[0071] Figure 4 This is a schematic diagram showing an example of the non-combustion heating type flavor inhalation system according to the present embodiment.

[0072] Figure 5 Schematic diagram showing an example of the non-combustion and non-heating type flavor inhaler according to the present embodiment.

[0073] Figure 6 This is a schematic diagram showing an example of a tobacco capsule of the non-combustion and non-heating type flavor inhaler of the present embodiment.

[0074] Figure 7 1 is a schematic diagram showing an example of a power supply unit of the non-combustion and non-heating type flavor inhaler according to the present embodiment.

[0075] Figure 8 This is a schematic diagram showing an example of a cartridge of the non-combustion and non-heating type flavor inhaler of the present embodiment.

[0076] Fig. 9 This is a schematic diagram showing an example of a cartridge of the non-combustion and non-heating type flavor inhaler of the present embodiment.

[0077] Fig.10 It is a schematic diagram showing an example of the combustion type flavor inhaler according to the present embodiment.

[0078] Fig.11 This is a total ion chromatogram obtained by GC / MS analysis of a n-hexane solution in Reference Example 2.

[0079] Fig.12 This is a total ion chromatogram obtained by GC / MS analysis of the ethyl acetate solution in Reference Example 2.

[0080] Fig.13 This is a total ion chromatogram obtained by GC / MS analysis of an ethanol solution in Reference Example 2.

[0081] Fig.14 This is a total ion chromatogram obtained by GC / MS analysis of a methanol solution in Reference Example 2.

[0082] Fig.15This is a total ion chromatogram obtained by GC / MS analysis of the solution obtained in step B1 of Example 1.

[0083] Fig.16 This is a total ion chromatogram obtained by GC / MS analysis of the hexane solution obtained in step C1 of Example 1.

[0084] Fig.17 This is a total ion chromatogram obtained by GC / MS analysis of the methanol solution obtained in step C1 of Example 1.

[0085] Fig.18 This is a total ion chromatogram obtained by GC / MS analysis of the supernatant obtained in step F1 of Example 1.

[0086] Fig.19 This is a chromatogram obtained by fractionating neophytadiene by HPLC in step H1 of Example 1.

[0087] Fig. 20 This is a total ion chromatogram obtained by subjecting the solution of the neophytadiene composition obtained in Example 1 to GC / MS analysis.

[0088] Explanation of symbols

[0089] 10Heating device

[0090] 11 Body

[0091] 12 Heater

[0092] 13 Metal pipe

[0093] 14 battery cells

[0094] 15Control Unit

[0095] 16 recesses

[0096] 17 Ventilation holes

[0097] 20 Non-combustion heating type aroma inhaler

[0098] 20A cigarette holder

[0099] 20B Cooling unit

[0100] 20C filter part

[0101] 21 Tobacco filler

[0102] 22 rolls of paper

[0103] 23 Paper tube

[0104] 24 piercing

[0105] 25 Section 1

[0106] 25a 1st filling layer

[0107] 25b inner rod packaging material

[0108] 26 Section 2

[0109] 26a Second filling layer

[0110] 26b inner rod packaging material

[0111] 27 Outer rod packaging material

[0112] 28 backing paper

[0113] 30 non-burning and non-heating aroma inhalers

[0114] 30D Power Supply Unit

[0115] 30E cigarette cartridge

[0116] 30F tobacco capsule

[0117] Non-suction end

[0118] d Suction port

[0119] 110 battery

[0120] 200 Aerosol Source

[0121] 210 cigarette storage

[0122] 220 Atomization Department

[0123] 230 Channel Forming Body

[0124] 240 outer frame 240

[0125] 250 end cap

[0126] 200X Channel 1

[0127] 300 scent sources

[0128] 310 container

[0129] 320 mesh body

[0130] 330 non-woven fabric

[0131] 340 cover

[0132] 400 Burning Fragrance Inhaler

[0133] 401 Contains tobacco section

[0134] 402 filter section

[0135] 403 Tobacco Filler

[0136] 404 roll of paper

[0137] 405 Tipping Paper Components DETAILED DESCRIPTION

[0138] [Method for separating and purifying neophytadiene from tobacco leaves]

[0139] <First Embodiment>

[0140] The method for separating and purifying neophytadiene from tobacco leaves according to the present embodiment includes the following steps.

[0141] A step of adding an aprotic solvent to tobacco leaves for extraction, and removing the extraction residue to obtain an extract (A1) (hereinafter also referred to as step A1);

[0142] 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);

[0143] 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);

[0144] A step of removing the solvent from the extract (C1) to obtain a dry solid (D1) (hereinafter also referred to as step D1);

[0145] A step of adding a protic solvent to the dry solid (D1) to dissolve the dry solid (D1) to obtain a solution (E1) (hereinafter also referred to as step E1);

[0146] A step of removing the precipitate generated in the solution (E1) to obtain a supernatant (F1) (hereinafter also referred to as step F1);

[0147] A step of replacing the solvent of the supernatant (F1) with an aprotic solvent to obtain a solution (G1) (hereinafter also referred to as step G1); and

[0148] A step of separating and purifying neophytadiene from the solution (G1) by normal phase high performance liquid chromatography (hereinafter also referred to as step H1).

[0149] The method of the present embodiment includes the above-mentioned steps A1 to H1, and can separate and purify neophytadiene from tobacco leaves with high yield and high purity. Generally, it is not possible to completely separate neophytadiene and nicotine in normal phase high performance liquid chromatography, so it is difficult to separate and purify neophytadiene with high yield and high purity. However, in the method of the present embodiment, in particular, for the extract obtained from tobacco leaves through step A1, the alkaloids with nicotine as the main component contained in the above-mentioned extract are first separated by adding an acidic aqueous solution and removing the water layer through step B1. Therefore, since nicotine and the like are removed at the beginning, the yield and purity of neophytadiene after separation and purification can be efficiently improved. Furthermore, after the solvent of the extract (C1) is intentionally removed through step D1 to obtain a dry solid (D1), a protic solvent can be added through step E1 to dissolve the above-mentioned dry solid (D1) again, and the precipitate (higher hydrocarbons) generated in the obtained solution (E1) is removed through step F1, thereby efficiently separating higher hydrocarbons. Therefore, the yield and purity of the isolated and purified neophytadiene can be more efficiently improved. Below, steps A1 to H1 are described in detail. The method of this embodiment may also include other steps in addition to steps A1 to H1. A flow chart of an example of the method of this embodiment is shown in Figure 1 .

[0150] (Process A1)

[0151] In this step, an aprotic solvent is added to the tobacco leaves to perform extraction, and the extraction residue is removed to obtain an extract (A1). Through this step, the effective components containing neophytadiene contained in the tobacco leaves can be extracted into the extract (A1).

[0152] There is no particular limitation on the type of tobacco leaves, for example, yellow tobacco, burley tobacco, oriental tobacco, local varieties (incoming varieties), and other Nicotiana tabacum varieties, and Nicotiana rustica varieties. One of these tobacco leaf varieties can be used, or two or more can be used in combination. Among them, from the viewpoint of containing more neophytadiene, it is preferably at least one tobacco leaf selected from the yellow type and burley tobacco, and the yellow type is more preferred. There is no particular limitation on the shape of the tobacco leaves, and from the viewpoint of improving the extraction efficiency, fine powdered tobacco leaves are preferably used. It should be noted that as tobacco leaves, solids discharged as waste from the expansion process of tobacco leaves and tobacco leaf fine powder discharged from a tobacco leaf raw material factory can also be used.

[0153] From the viewpoint of being able to more selectively extract neophytadiene, the aprotic solvent used as the extraction solvent is preferably at least one solvent selected from hexane, heptane and ethyl acetate, and more preferably hexane. The amount of the aprotic solvent added is preferably 500 to 2000 parts by mass, more preferably 750 to 1250 parts by mass, relative to 100 parts by mass of tobacco leaves. The extraction operation can be carried out by stirring the aprotic solvent containing the tobacco leaves, etc. The extraction temperature is preferably 15 to 40°C, and the extraction time is preferably 1 to 6 hours. The removal of the extraction residue can be carried out, for example, by separating the extract (A1) and the extraction residue from each other using a stainless steel sieve.

[0154] (Process B1)

[0155] In this step, an acidic aqueous solution is added to the extract (A1) obtained in the above step A1 to perform liquid-liquid extraction, and the aqueous layer is removed to obtain an extract (B1). In this step, alkaloids with nicotine as the main component are transferred to the aqueous layer, while hydrophobic active ingredients of tobacco leaves such as neophytadiene remain in the organic layer, so that alkaloids with nicotine as the main component can be selectively separated and removed.

[0156] In the present embodiment, the acidic aqueous solution refers to an aqueous solution having a pH lower than 7. The acidic aqueous solution may be, for example, an aqueous solution containing a pH adjuster such as sulfuric acid or hydrochloric acid. From the viewpoint of making it easier to transfer alkaloids represented by nicotine to the water layer, the pH of the acidic aqueous solution is preferably 3 or less, more preferably 1 to 3. It should be noted that in the present specification, the pH of the aqueous solution is a value measured by a pH meter (e.g., trade name: desktop pH meter, manufactured by HORIBA).

[0157] The amount of the acidic aqueous solution added also 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 and the like. The extraction temperature is preferably 15 to 40°C, and the extraction time is preferably 0.5 to 3 hours. In addition, a salt such as sodium chloride can also be added to the mixed solution during the liquid-liquid extraction. A desiccant such as anhydrous sodium sulfate can also be added to the extract (B1) obtained by separating and removing the water layer to perform a dehydration operation in the extract (B1).

[0158] (Process C1)

[0159] In this step, a protic solvent is added to the extract (B1) obtained in the above step B1 to perform liquid-liquid extraction, and the protic solvent layer is removed to obtain an extract (C1). Through this step, the tobacco leaf surface resin component containing cebutriene glycol (CBT) as the main component is transferred to the protic solvent layer, and the neophytadiene and higher hydrocarbons are each transferred to the aprotic solvent layer, so that the tobacco leaf surface resin component containing CBT as the main component can be selectively separated and removed.

[0160] As a protic solvent, methanol is preferred from the viewpoint of being able to more selectively extract and remove the leaf surface resin components of tobacco leaves with CBT as the main component. As for the amount of protic solvent added, the volume ratio of the extract (B1) and the protic solvent (extract (B1): protic solvent) is preferably 100:15-100, more preferably 100:40-60. The liquid-liquid extraction operation can be performed by shaking the mixed solution and the like. The extraction temperature is preferably 15-40°C, and the extraction time is preferably 3-8 hours. In addition, from the viewpoint of further removing the leaf surface resin components of tobacco leaves with CBT as the main component, this process can be repeated twice or more.

[0161] (Process D1)

[0162] In this step, the solvent of the extract (C1) obtained in the above step C1 is removed to obtain a dry solid (D1). The method for removing the solvent (aprotic solvent) of the extract (C1) is not particularly limited, and for example, the solvent can be removed from the extract (C1) under reduced pressure.

[0163] (Process E1)

[0164] In this step, a protic solvent is added to the dry solid (D1) obtained by the above step D1 to dissolve the above dry solid (D1) to obtain a solution (E1). As a protic solvent, methanol is preferred from the viewpoint that higher hydrocarbons are difficult to dissolve and neophytadiene is easy to dissolve. As the amount of the protic solvent added, the protic solvent is preferably 750 to 2500 parts by mass, and more preferably 1000 to 1500 parts by mass, relative to 100 parts by mass of the dry solid (D1). There is no particular limitation on the method for dissolving the dry solid (D1) in the protic solvent. For example, the dry solid (D1) can be dispersed and dissolved by ultrasonic treatment of the solution. It should be noted that the dry solid (D1) does not need to be completely dissolved in the protic solvent, and only needs to be at least partially dissolved.

[0165] (Process F1)

[0166] In this step, the precipitate generated in the solution (E1) obtained in the above step E1 is removed to obtain a supernatant (F1). Through this step, higher hydrocarbons as precipitates can be selectively separated and removed. The precipitate can be generated by leaving the solution (E1) to stand. The standing can be performed at 0 to 30° C. for 1 to 12 hours, for example.

[0167] (Process G1)

[0168] In this step, the solvent of the supernatant (F1) obtained in the above step F1 is replaced with an aprotic solvent to obtain a solution (G1). In this step, the solvent (protic solvent) of the supernatant (F1) is replaced with an aprotic solvent. The method is not particularly limited. For example, an aprotic solvent may be added to the supernatant (F1) to perform liquid-liquid extraction, and the solution (G1) may be obtained by removing the protic solvent layer. Alternatively, an aprotic solvent may be added after removing the solvent of the supernatant (F1) to obtain the solution (G1).

[0169] (Process H1)

[0170] In this operation, by normal phase high performance liquid chromatography (normal phase HPLC), the solution (G1) obtained from the above-mentioned operation G1 is separated and purified. It is not particularly limited as HPLC, for example, 1260 Infinity (trade name, Agilent Technologies company system) in the commercially available product can be used. When using the above-mentioned HPLC, separation operation can be carried out by, for example, the following conditions.

[0171] ·Chromatographic column: YMC-Pack SIL / S-5μm / 12nm manufactured by YMC Corporation

[0172] Mobile phase: Hex (2 min) → Hex: AcOEt (95:5) (1 min) → Hex (12 min)

[0173] Flow rate: 7mL / min

[0174] Detector: Diode array detector (220nm)

[0175] Sample injection volume: 500μL

[0176] After separation under the above conditions, the signal detected at 220nm is graded in a fraction collector, and the graded fractions are recovered, and the separation and purification of neophytadiene can be carried out by concentrating the obtained sample, drying and solidifying. Neophytadiene can be obtained in high yield by the method of this embodiment, and the neophytadiene composition obtained by separation and purification contains more than 95% by mass of neophytadiene.

[0177] <Second Embodiment>

[0178] The method for separating and purifying neophytadiene from tobacco leaves according to the present embodiment includes the following steps.

[0179] A step of adding an aprotic solvent to tobacco leaves for extraction, and removing the extraction residue to obtain an extract (A2) (hereinafter also referred to as step A2);

[0180] 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);

[0181] A step of removing the solvent from the extract (B2) to obtain a dry solid (C2) (hereinafter also referred to as step C2);

[0182] A step of adding a protic solvent to the dry solid (C2) to dissolve the dry solid (C2) to obtain a solution (D2) (hereinafter also referred to as step D2);

[0183] A step of removing the precipitate generated in the solution (D2) to obtain a supernatant (E2) (hereinafter also referred to as step E2);

[0184] A step of adding an aprotic solvent to the supernatant (E2) to perform liquid-liquid extraction, removing the protic solvent layer, and obtaining a solution (F2) (hereinafter also referred to as step F2);

[0185] A step of separating and purifying neophytadiene from the solution (F2) by normal phase high performance liquid chromatography (hereinafter also referred to as step G2).

[0186] The method of the present embodiment can separate and purify neophytadiene from tobacco leaves with high purity by including the above-mentioned steps A2 to G2. In the method of the present embodiment, in particular, the extract obtained from tobacco leaves by step A2 is first subjected to step B2 to add an acidic aqueous solution and remove the water layer, thereby separating the alkaloids with nicotine as the main component contained in the above-mentioned extract. Therefore, since nicotine and the like are removed initially, the yield and purity of the neophytadiene after separation and purification can be efficiently improved. Furthermore, after the solvent of the extract (B2) is intentionally removed by step C2 to obtain a dry solid (C2), a protic solvent can be added by step D2 to dissolve the above-mentioned dry solid (C2) again, and the precipitate (higher hydrocarbons) generated in the obtained solution (D2) can be removed by step E2, so that higher hydrocarbons can be efficiently separated. Therefore, the yield and purity of the neophytadiene after separation and purification can be more efficiently improved. Below, steps A2 to G2 are described in detail, and the method of the present embodiment can also include other steps in addition to steps A2 to G2. A flowchart of an example of the method of this embodiment is shown in Figure 2 .

[0187] (Process A2, B2)

[0188] Steps A2 and B2 can be carried out in the same manner as steps A1 and B1 in the first embodiment.

[0189] (Process C2)

[0190] In this step, the solvent of the extract (B2) obtained in the above step B2 is removed to obtain a dry solid (C2). The method for removing the solvent (aprotic solvent) of the extract (B2) is not particularly limited, and for example, the solvent can be removed from the extract (B2) under reduced pressure.

[0191] (Process D2)

[0192] In this step, a protic solvent is added to the dry solid (C2) obtained by the above step C2 to dissolve the above dry solid (C2) to obtain a solution (D2). As a protic solvent, methanol is preferably used from the viewpoint that higher hydrocarbons are difficult to dissolve and neophytadiene is easy to dissolve. As the amount of the protic solvent added, the protic solvent is preferably 500 to 2500 parts by mass, and more preferably 750 to 1500 parts by mass, relative to 100 parts by mass of the dry solid (C2). There is no particular limitation on the method for dissolving the dry solid (C2) in the protic solvent. For example, the solution can be ultrasonically treated to disperse and dissolve the dry solid (C2). It should be noted that the dry solid (C2) does not need to be completely dissolved in the protic solvent, only needs to be at least partially dissolved.

[0193] (Process E2)

[0194] In this step, a supernatant (E2) is obtained by removing the precipitate generated in the solution (D2) obtained in the above step D2. By this step, higher hydrocarbons can be selectively separated and removed as precipitates. The precipitate can be generated by leaving the solution (D2) to stand. The standing can be performed at 15 to 40° C. for 3 to 8 hours, for example.

[0195] (Process F2)

[0196] In this step, an aprotic solvent is added to the supernatant (E2) obtained in the above step E2 to perform liquid-liquid extraction, and the protic solvent layer is removed to obtain a solution (F2). Through this step, the tobacco leaf surface resin component containing cebutriene glycol (CBT) as the main component is transferred to the protic solvent layer, and neophytadiene is transferred to the aprotic solvent layer, so that the tobacco leaf surface resin component containing CBT as the main component can be selectively separated and removed.

[0197] As an aprotic solvent, from the viewpoint of being able to more selectively extract neophytadiene, it is preferred to use at least one solvent selected from hexane, heptane and ethyl acetate, and more preferably hexane. As the amount of aprotic solvent added, the volume ratio of the supernatant (E2) and the aprotic solvent (supernatant (E2): aprotic solvent) is preferably 15 to 100:100, and more preferably 40 to 60:100. Liquid-liquid extraction can be performed by oscillating the mixed solution. The extraction temperature is preferably 0 to 30°C, and the extraction time is preferably 3 to 12 hours. In addition, from the viewpoint of further removing the leaf surface resin component of tobacco leaves with CBT as the main component, this process can be repeated more than twice.

[0198] (Process G2)

[0199] Step G2 can be carried out in the same manner as step H1 in the first embodiment. As in the first embodiment, the method of this embodiment can obtain neophytadiene at a high yield, and the neophytadiene composition obtained by separation and purification contains 95% by mass or more of neophytadiene.

[0200] [New phytadiene composition]

[0201] The neophytadiene composition of the present embodiment is obtained by the method for separating and purifying neophytadiene from tobacco leaves of the present embodiment described above, and contains more than 95% by mass of neophytadiene. The neophytadiene composition preferably contains more than 97% by mass of neophytadiene, and more preferably contains more than 99% by mass of neophytadiene. It should be noted that the content (purity) of neophytadiene in the neophytadiene composition is a value measured by GC. In addition, the components other than neophytadiene contained in the neophytadiene composition obtained by the method of the present embodiment are mainly various components derived from tobacco leaves, and since they are only trace amounts, they are difficult to identify.

[0202] [Non-combustion heating type fragrance inhaler and its manufacturing method]

[0203] The manufacturing method of the non-combustion heating type flavor inhaler of the present embodiment includes: a process of separating and purifying neophytadiene from tobacco leaves by the method of the present embodiment to obtain a neophytadiene composition; a process of adding the above-mentioned neophytadiene composition to a substrate; and a process of manufacturing a non-combustion heating type flavor inhaler containing the above-mentioned substrate. In addition, the non-combustion heating type flavor inhaler of the present embodiment contains the neophytadiene composition of the present embodiment. The manufacturing method of the non-combustion heating type flavor inhaler of the present embodiment obtains the neophytadiene composition by the method of the present embodiment, so that neophytadiene can be efficiently separated and purified from tobacco leaves, and a non-combustion heating type flavor inhaler that can provide good fragrance can be efficiently manufactured. In addition, since the non-combustion heating type flavor inhaler of the present embodiment contains the neophytadiene composition obtained by the method of the present embodiment, it can provide good fragrance.

[0204] (Base material)

[0205] In the method for manufacturing the non-combustion heating type flavor inhaler of the present embodiment, the base material to which the neophytadiene composition is added may be a tobacco material, such as a tobacco sheet, shredded tobacco, rolling paper, a polysaccharide sheet, and the like.

[0206] (1) Tobacco slices

[0207] Tobacco sheets are sheets obtained by molding a composition including cured tobacco leaves and the like. The cured tobacco leaves used in the tobacco sheets are not particularly limited, and for example, tobacco leaves that have been destemmed and separated into lamina and veins can be cited. Cured tobacco leaves refer to tobacco leaves that have been subjected to treatments such as baking and have been stored for a long time in a warehouse or the like. The "sheet" in this embodiment refers to a material having a pair of roughly parallel main surfaces and side surfaces. Tobacco sheets can be molded by known methods such as papermaking, casting, and rolling. Various tobacco sheets molded by such methods are disclosed in detail in "Tobacco Encyclopedia, Comprehensive Tobacco Research Center, 2009.3.31". The manner of adding the new phytadiene composition of this embodiment to the tobacco sheets is not limited.

[0208] For example, the neophytadiene composition of the present embodiment can be dissolved in a solvent to prepare a tobacco flavoring agent in a liquid state, and the flavoring agent can be sprayed or impregnated into a finished tobacco sheet, or the neophytadiene composition of the present embodiment can be added when the tobacco sheet is formed. For example, in the papermaking method, the following steps are taken: extracting water-soluble components from the cured tobacco leaves and separating them into a water extract and a residue, making paper with a mixture of the fibrous residue and the slurry, and adding a concentrated solution of the water extract to the sheet after papermaking, and the neophytadiene composition of the present embodiment can be added to the water extract. In the casting method, the neophytadiene composition of the present embodiment can be added to the mixture by mixing water, slurry, adhesive and a crushed material of the cured tobacco to form a mixture and casting it. In the rolling method, the neophytadiene composition of the present embodiment can be added to the mixture by mixing water, slurry, adhesive and a crushed material of the cured tobacco to form a mixture and putting it into a plurality of rollers for rolling.

[0209] Further, as described in International Publication No. 2014 / 104078, the cured tobacco pulverized material is mixed with a binder to form a mixture, the mixture is clamped by a non-woven fabric, and then the laminate is formed into a certain shape by heat fusion, so that a non-woven fabric-like tobacco sheet can be obtained. In this method, the new phytadiene composition of the present embodiment can be added to the above-mentioned mixture.

[0210] The tobacco sheet may include an aerosol-generating substrate. The type of aerosol-generating substrate is not particularly limited, and extracts from various natural products or their constituent components can be selected according to the purpose. As specific examples of aerosol-generating substrates, polyols such as glycerol, propylene glycol, sorbitol, xylitol, and erythritol, triacetin, 1,3-butylene glycol, and mixtures thereof can be cited. The content of the aerosol-generating substrate can be adjusted to various amounts according to the form of use in tobacco products. For example, when an aerosol-generating substrate is included in a tobacco sheet, from the viewpoint of obtaining a good flavor, its content is generally 5% by mass or more, preferably 10% by mass or more, and more preferably 15% by mass or more, relative to the total mass of the tobacco sheet, and is generally 50% by mass or less, preferably 40% by mass or less, and more preferably 25% by mass or less.

[0211] (2) Tobacco

[0212] As shredded tobacco, there can be cited: shreds obtained by shredding aged tobacco leaves into a given size, shreds obtained by shredding the above-mentioned tobacco sheets into a given size, or shreds obtained by mixing them. The size is not limited, and as an example, shreds with a width of 0.5 to 2.0 mm and a length of 3 to 10 mm can be cited. A preferred embodiment is to fill shredded tobacco of such a size into the filler described later. In addition, as shredded tobacco, there can be cited shredded tobacco leaves after processing into strands of shreds with a width of 0.5 to 2.0 mm and a length longer than the above-mentioned shredded tobacco, preferably the same horizontal length as the rolling paper. The new phytadiene composition of this embodiment can be added to shredded tobacco, or it can be added to the raw material before shredding.

[0213] The shredded tobacco may contain the above-mentioned aerosol generating substrate. When the shredded tobacco contains the aerosol generating substrate, from the viewpoint of generating a sufficient amount of aerosol while also obtaining a good flavor, the content thereof is usually 5% by mass or more, preferably 10% by mass or more, more preferably 15% by mass or more, and is usually 50% by mass or less, preferably 40% by mass or less, more preferably 25% by mass or less, relative to the mass of the shredded tobacco.

[0214] (3) Toilet paper

[0215] The rolling paper containing the neophytadiene composition can be prepared by dissolving the neophytadiene composition of the present embodiment in a solvent, preparing a solution-like tobacco flavoring agent, spraying the flavoring agent, impregnating the rolling paper, etc. As the rolling paper, for example, a material with pulp as the main component can be cited. As the pulp, in addition to papermaking with wood pulp such as conifer pulp and broadleaf pulp, non-wood pulp generally used for rolling paper for tobacco products such as flax pulp, sisal pulp, Spanish grass pulp, etc. can also be mixed with papermaking and manufactured to obtain. These pulps can be used alone or in combination in any proportion. In addition, the rolling paper can be composed of one sheet or more than one sheet. The rolling paper can be used for the method of wrapping tobacco raw materials such as shredded tobacco, and can also be used as a material (for example, tipping paper) for wrapping the wrapped component with other components such as cooling components and filter components. As pulp, chemical pulp obtained by kraft cooking, acid / neutral / alkaline sulfite cooking, sodium carbonate cooking, etc., refined pulp, chemical mechanical pulp, thermomechanical pulp, etc. can be used.

[0216] (4) Polysaccharide tablets

[0217] The polysaccharide sheet refers to a sheet with polysaccharides as the main component, and the neophytadiene composition of the present embodiment can be contained in the polysaccharide sheet. The non-combustion heating type fragrance inhaler using the polysaccharide sheet containing the neophytadiene composition of the present embodiment can release a good fragrance. As polysaccharides, carrageenan, agar, gellan gum, tamarind gum, psyllium seed gum, konjac glucomannan, carrageenan, locust bean gum, guar gum, agar, xanthan gum, gellan gum, tamarind gum, thorny tava gum, konjac glucomannan, starch, cassia gum, and psyllium seed gum can be cited.

[0218] The polysaccharide sheet containing the neophytadiene composition of the present embodiment can be used for a non-combustion heating type flavor inhaler, and a non-combustion non-heating type flavor inhaler and a combustion type flavor inhaler described later. In the embodiment of the combustion type flavor inhaler, for example, a polysaccharide sheet as disclosed in Patent No. 5481574 can be used. In the present embodiment, the content of the neophytadiene composition of the present embodiment can be 0.01 to 30% by mass relative to the sheet. The polysaccharide sheet can be prepared by the following method: a polysaccharide is mixed with water, heated to prepare an aqueous solution of the polysaccharide, and then spices and emulsifiers are added to the aqueous solution to mix and emulsify it. As an emulsifier, a known emulsifier can be used.

[0219] In the embodiment of the non-combustion heating type flavor inhaler, a polysaccharide sheet as described in PCT / JP2019 / 20136 can be used. In the present embodiment, agar is particularly preferably used as a polysaccharide. Relative to the sheet, the content of agar is preferably 10 to 50% by mass, more preferably 15 to 45% by mass. In addition, in the polysaccharide sheet, the neophytadiene composition of the present embodiment can be 0.01 to 30% by mass relative to the sheet.

[0220] In the present embodiment, it is preferred to use a carbohydrate compound selected from sugars and sugar alcohols. As "sugar", for example, glucose, sucrose, fructose, xylose, galactose, mannose, maltose, trehalose, lactose, and raffinose can be mentioned. As "sugar alcohol", sorbitol, which is an alcohol obtained by reducing the carbonyl group of sugar to a hydroxyl group, can be mentioned. Relative to agar, the content of the compound is preferably 10% by mass or more, more preferably 10 to 500% by mass, further preferably 10 to 300% by mass, and further preferably 10 to 200% by mass. In addition, in the present embodiment, an emulsifier is preferably used. As an emulsifier, a known emulsifier can be used, and its content is preferably 0.5 to 10% by mass, more preferably 1.0 to 8.0% by mass relative to the mass of agar.

[0221] The polysaccharide sheet in the present embodiment can be produced by kneading raw materials containing agar, a saccharide compound, a flavor, and an emulsifier in water to prepare a raw material slurry, spreading the raw material slurry on a substrate, and drying the raw material slurry.

[0222] (Non-combustion heating type aroma inhaler)

[0223] In the present embodiment, the "flavor inhaler" refers to an article used by the user to inhale the fragrance. Fragrance inhalers are roughly divided into "combustion type flavor inhalers" that produce fragrance by combustion, and "non-combustion type flavor inhalers" that do not produce fragrance by combustion. Furthermore, non-combustion type flavor inhalers are roughly divided into "non-combustion heating type flavor inhalers" that produce fragrance by heating, and "non-combustion non-heating type flavor inhalers" that produce fragrance without heating. In addition, the combination of a device for generating aerosol (a heating device or an atomizing device, etc.) and a non-combustion heating type flavor inhaler is particularly referred to as a non-combustion heating type flavor inhalation system.

[0224] exist Figure 3 One embodiment of the non-combustion heating type flavor inhaler of this embodiment is shown in FIG. Figure 3 As shown, the non-combustion heating type flavor inhaler 20 comprises: a cigarette rod portion 20A, a cylindrical cooling portion 20B having perforations on the circumference, and a filter portion 20C. The non-combustion heating type flavor inhaler 20 may also have components other than these. The axial length of the non-combustion heating type flavor inhaler 20 is not limited, and is preferably 40 to 90 mm, more preferably 50 to 75 mm, and further preferably 50 to 60 mm or less. In addition, the circumference of the non-combustion heating type flavor inhaler 20 is preferably 16 to 25 mm, more preferably 20 to 24 mm, and further preferably 21 to 23 mm. For example, a method in which the length of the cigarette rod portion 20A is 20 mm, the length of the cooling portion 20B is 20 mm, and the length of the filter portion 20C is 7 mm can be cited. The lengths of these individual components can be appropriately changed according to manufacturing adaptability, required quality, etc. Figure 3 2 shows an embodiment in which the first section 25 is arranged, but this section may not be arranged and the second section 26 may be arranged only on the downstream side of the cooling unit 20B.

[0225] (1) Cigarette rod 20A

[0226] In the cigarette rod portion 20A, as the tobacco filler 21, shredded tobacco or tobacco sheets containing the neophytadiene composition of the present embodiment can be used. The method of filling the tobacco filler 21 into the rolling paper 22 is not particularly limited. For example, the tobacco filler 21 can be wrapped with the rolling paper 22, or the tobacco filler 21 can be filled into the cylindrical rolling paper 22. When the shape of the tobacco is rectangular with a long direction, it can be filled in a manner that the long direction is an unspecified direction in the rolling paper 22, or it can be filled in a manner that is arranged in the axial direction of the cigarette rod portion 20A or in a direction orthogonal thereto. In addition, as the rolling paper 22, the rolling paper containing the neophytadiene composition of the present embodiment described above can also be used. Since the cigarette rod portion 20A is heated, the tobacco components, aerosol generating substrate and water contained in the tobacco filler 21 are vaporized for inhalation.

[0227] (2) Cooling section 20B

[0228] The cooling section 20B is preferably composed of a tubular member. The tubular member may be, for example, a paper tube 23 obtained by processing thick paper into a cylindrical shape. In addition, in order to form a channel, the cooling section 20B may be formed by a sheet of a wrinkled, pleated, pleated, or folded thin material. As such a material, for example, a sheet material selected from polyethylene, polypropylene, polyvinyl chloride, polyethylene terephthalate, polylactic acid, cellulose acetate, and aluminum foil may be used. The total surface area of ​​the cooling section 20B is appropriately adjusted in consideration of the cooling efficiency, and may be, for example, 300 to 1000 mm 2 / mm. It is preferred to provide a perforation 24 on the cooling portion 20B. Due to the presence of the perforation 24, external air is introduced into the cooling portion 20B during inhalation. Therefore, by heating the cigarette rod portion 20A, the vaporized component of the generated aerosol comes into contact with the external air, and its temperature drops, resulting in liquefaction to form an aerosol. The diameter (diameter length) of the perforation 24 is not particularly limited, and may be, for example, 0.5 to 1.5 mm. The number of perforations 24 is not particularly limited, and may be 1 or more than 2. For example, a plurality of perforations 24 may be provided on the circumference of the cooling portion 20B.

[0229] The axial length of the cooling portion 20B may be, for example, a rod-shaped portion of 7 to 28 mm. For example, the axial length of the cooling portion 20B may be 18 mm. The axial cross-sectional shape of the cooling portion 20B is substantially circular, and the diameter may be 5 to 10 mm. For example, the diameter of the cooling portion may be about 7 mm.

[0230] (3) Filter part 20C

[0231] The structure of the filter portion 20C is not particularly limited, and may be composed of a single or multiple filling layers. The outer side of the filling layer may be wrapped with one or more sheets of toilet paper. The ventilation resistance of the filter portion 20C may be appropriately changed by the amount and material of the filler filled in the filter portion 20C. For example, when the filler is cellulose acetate fiber, the ventilation resistance may be increased by increasing the amount of cellulose acetate fiber filled in the filter portion 20C. When the filler is cellulose acetate fiber, the filling density of the cellulose acetate fiber may be 0.13 to 0.18 g / cm 3 The ventilation resistance is a value measured by a ventilation resistance measuring instrument (trade name: SODIMAX, manufactured by SODIM).

[0232] 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 still more preferably 21 to 23 mm. Figure 3 The horizontal length can be selected from 4 to 10 mm, and the ventilation resistance is 15 to 60 mmH. 2 The filter portion 20C is preferably selected in the form of O / seg. The axial length of the filter portion 20C is preferably 5 to 9 mm, more preferably 6 to 8 mm. The shape of the cross section of the filter portion 20C is not particularly limited, and may be, for example, circular, elliptical, polygonal, etc. In addition, destructive capsules, flavor beads, and flavors containing flavors may be directly added to the filter portion 20C.

[0233] As the first section 25, the filter portion 20C may have a central hole portion. The central hole portion is composed of a first filling layer 25a having one or more hollow portions, and an inner rod wrapping material (inner wrapping paper) 25b covering the filling layer. The central hole portion has the function of increasing the strength of the mouthpiece portion. The central hole portion may also not have an inner rod wrapping material 25b and maintain its shape by thermoforming. The first filling layer 25a may be, for example, a rod with an inner diameter of φ5.0 to φ1.0 mm, which is filled with cellulose acetate fibers at a high density and a plasticizer containing triacetin is added at 6 to 20% by mass relative to the mass of cellulose acetate and solidified. Since the fiber filling density of the first filling layer 25a is high, air and aerosol only flow through the hollow portion during inhalation, and basically do not flow through the first filling layer 25a. Since the first filling layer 25a of the central hole portion is a fiber filling layer, the touch from the outside during use will basically not make the user feel uncomfortable. The filter portion 20C may include a second section 26. The second section 26 is composed of a second filling layer 26a and an inner rod wrapper (inner wrapping paper) 26b covering the filling layer.

[0234] The first section 25 and the second section 26 are connected by an outer rod packaging material (outer winding paper) 27. The outer rod packaging material 27 can be, for example, a cylindrical paper. In addition, the cigarette rod portion 20A, the cooling portion 20B, the connected first section 25 and the second section 26 are connected by the cigarette holder lining paper 28. These connections can be made by, for example, applying a paste such as vinyl acetate paste on the inner side surface of the cigarette holder lining paper 28 and winding the above three components. These components can also be connected in multiple times using multiple lining papers.

[0235] (Non-combustion heating type aroma inhalation system)

[0236] An example of the non-combustion heating type flavor inhalation system of this embodiment is shown in Figure 4 . Figure 4 In the embodiment, the non-combustion heating type flavor inhalation system includes: a non-combustion heating type flavor inhalation device 20 and a heating device 10 for heating the cigarette rod portion 20A from the outside.

[0237] 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 the metal tube 13 are arranged at a position corresponding to the cigarette rod portion 20A inserted therein. The heater 13 can be a heater using resistance, and the control unit 15 for temperature control issues instructions, and the battery unit 14 supplies power to heat the heater 12. The heat generated by the heater 12 can be conducted to the cigarette rod portion 20A through the metal tube 13 with high thermal conductivity. Figure 4 , the heating device 10 heats the cigarette rod 20A from the outside, but the heating may also be performed from the inside. The temperature of heating by the heating device 10 is not particularly limited, but is preferably 400° C. or less, more preferably 150 to 400° C., and further preferably 200 to 350° C. The heating temperature refers to the temperature of the heater 12 of the heating device 10.

[0238] [Non-burning and non-heating type fragrance inhaler and its manufacturing method]

[0239] According to the method of the present embodiment, the manufacturing method of the non-burning non-heating flavor inhaler of the present embodiment includes: a process of separating and purifying neophytadiene from tobacco leaves to obtain a neophytadiene composition, a process of adding the above-mentioned neophytadiene composition to a substrate, and a process of manufacturing a non-burning non-heating flavor inhaler comprising the above-mentioned substrate. In addition, the non-burning non-heating flavor inhaler of the present embodiment includes the neophytadiene composition of the present embodiment. The manufacturing method of the non-burning non-heating flavor inhaler of the present embodiment obtains the neophytadiene composition by the method of the present embodiment, so that the neophytadiene can be efficiently separated and purified from tobacco leaves, and the non-burning non-heating flavor inhaler that can provide good fragrance can be efficiently manufactured. In addition, since the non-burning non-heating flavor inhaler of the present embodiment includes the neophytadiene composition obtained by the method of the present embodiment, a good fragrance can be provided.

[0240] In the method for producing the non-combustion and non-heating type flavor inhaler of the present embodiment, the method for adding the neophytadiene composition to the base material can be performed in the same manner as the method for producing the non-combustion and heating type flavor inhaler described above.

[0241] Figure 5 One mode of the non-burning and non-heating flavor inhaler of the present embodiment is shown. The non-burning and non-heating flavor inhaler 30 has a power supply unit 30D, a cigarette cartridge 30E and a tobacco capsule 30F. The non-burning and non-heating flavor inhaler 30 has a shape extending from the non-inhalation end u (downstream) to the inhalation end d (downstream). The cigarette cartridge 30E is removable relative to the power supply unit 30D. In addition, the tobacco capsule 30F is removable relative to the cigarette cartridge 30E.

[0242] (1) Tobacco capsules

[0243] Figure 6 An example of a tobacco capsule 30F is shown. Figure 6 As shown, the tobacco capsule 30F is a tobacco rod portion, and has a flavor source 300 inside. The flavor source 300 includes a tobacco material containing the neophytadiene composition of this embodiment. The tobacco capsule 30F is connected to the tobacco cartridge 30E. Specifically, a portion of the tobacco capsule 30F is contained in the tobacco cartridge 30E.

[0244] The tobacco capsule 30F has a container 310, a mesh body 320, a nonwoven fabric 330 and a lid 340 for accommodating a flavor source 300. The aerosol atomized by the atomizing portion 220 described later is introduced into the container 310 through the mesh body 320, and is given aerosol with a flavor by contacting with the flavor source 300. Thereafter, the aerosol is inhaled by the user through the nonwoven fabric 330. Like this, the non-combustion non-heating type flavor inhaler 30 can just give aerosol a flavor without heating the flavor source 300. In addition, aerosol is not produced by the flavor source 300 basically.

[0245] In the flow direction of the aerosol, the length of the tobacco capsule 30F (accommodating body 310) is preferably 40 mm or less, more preferably 25 mm or less. Further, in the flow direction of the aerosol, the above length is preferably 1 mm or more, more preferably 5 mm or more. In the direction orthogonal to the flow direction of the aerosol, the maximum length of the accommodating body 310 of the tobacco capsule 30F (accommodating body 310) is preferably 20 mm or less, more preferably 10 mm or less. Further, in the direction orthogonal to the flow direction of the aerosol, the maximum length is preferably 1 mm or more, more preferably 3 mm or more.

[0246] The flavor source 300 containing tobacco is composed of a raw material sheet that imparts flavor to the aerosol. The lower limit of the size of the raw material sheet is preferably 0.2 to 1.2 mm, more preferably 0.2 to 0.7 mm. The smaller the size of the raw material sheet constituting the flavor source 300, the larger the specific surface area, and thus it is easier to release flavor components. As the raw material sheet constituting the flavor source 300, cut tobacco containing the neophytadiene composition of the present embodiment, or a molded body obtained by molding a tobacco material containing the neophytadiene composition of the present embodiment into a granular shape can be used. The flavor source 300 may also contain plants other than tobacco (for example, mint, herb, etc.), spices such as menthol. Further, the flavor source 300 containing tobacco may also contain a flavoring agent. As the flavoring agent, for example, raw materials that exhibit sweetness, sourness, saltiness, umami, bitterness, astringency, richness, spiciness, pungency, astringent bitterness, etc. can be cited. As the raw material that exhibits sweetness, for example, saccharides, sugar alcohols, sweeteners, etc. can be cited. As saccharides, for example, monosaccharides, disaccharides, oligosaccharides, polysaccharides, etc. can be cited. As sweeteners, for example, natural sweeteners, synthetic sweeteners, etc. can be cited.

[0247] The raw material sheet can be obtained by sieving, for example, a stainless steel mesh conforming to JIS Z 8801 according to JIS Z8815. For example, a stainless steel mesh having a mesh size of 0.71 mm is used to sieve the raw material sheet by a dry and mechanical oscillation method for 20 minutes to obtain a raw material sheet that passes through the stainless steel mesh having a mesh size of 0.71 mm. Next, a stainless steel mesh having a mesh size of 0.212 mm is used to sieve the raw material sheet by a dry and mechanical oscillation method for 20 minutes to remove the raw material sheet that passes through the stainless steel mesh having a mesh size of 0.212 mm. That is, the raw material sheet constituting the flavor source 300 is a raw material sheet that passes through the stainless steel mesh having a prescribed upper limit (mesh size = 0.71 mm) and does not pass through the stainless steel mesh having a prescribed lower limit (mesh size = 0.212 mm). Therefore, the lower limit of the size of the raw material sheet constituting the flavor source 300 is defined according to the mesh size of the stainless steel mesh having a prescribed lower limit. In addition, the upper limit of the size of the raw material sheet constituting the flavor source 300 is defined by the mesh size of the stainless steel mesh that specifies the upper limit.

[0248] From the viewpoint of good flavor, the filling amount of the flavor source 300 contained in the container 310 is preferably 300 mg or more, more preferably 350 mg or more.

[0249] (2) Power supply unit

[0250] For example, a power supply unit 30D Figure 7 As shown. The power supply unit 30D has a battery 110. The battery 110 may be a disposable battery or a rechargeable battery. The initial value of the output voltage of the battery 110 is preferably in the range of 1.2V or more and 4.2V or less. In addition, the battery capacity of the battery 110 is preferably in the range of 100mAh or more and 1000mAh or less.

[0251] (3) Cigarette Cartridges

[0252] An example of a cigarette cartridge 30E is shown in Figure 8 , 9 . Figure 8 A diagram showing a cross-sectional view of an example of a cigarette cartridge 30E, Fig. 9 The cartridge 30E has a cigarette storage 210, an atomizing section 220, a channel forming body 230, an outer frame 240, and an end cap 250. The cartridge 30E has a first channel 200X provided on the downstream side of the atomizing section 220 as an aerosol channel.

[0253] The cigarette storage 210 stores the aerosol source 200. The cigarette storage 210 is located around the channel forming body 230 on a cross section perpendicular to the flow direction of the aerosol (the direction from the non-inhalation end toward the inhalation end (from downstream to downstream)). The cigarette storage 210 is located in the gap between the channel forming body 230 and the outer frame 240. The cigarette storage 210 is composed of a porous body such as a resin net, cotton, etc. In addition, the cigarette storage 210 can also be composed of a tank containing a liquid aerosol source 200. As the aerosol source 200, for example, glycerin, propylene glycol, etc. can be cited.

[0254] The atomizing section 220 atomizes the aerosol source 200 without combustion by means of the power supplied by the battery 110. The atomizing section 220 is composed of a heating wire (coil) wound at a given pitch. The atomizing section 220 is preferably composed of a heating wire having a resistance value in the range of 1.0 to 3.0 Ω. The given pitch is greater than a value at which the heating wires do not touch, and preferably a smaller value. The given pitch is, for example, preferably less than 0.40 mm. In order to stabilize the atomization of the aerosol source 200, the given pitch is preferably constant. The given pitch refers to the interval between the centers of adjacent heating wires.

[0255] The channel forming body 230 has a cylindrical shape that forms the first channel 200X that stretches along the flow direction of the aerosol. The outer frame 240 has a cylindrical shape that accommodates the channel forming body 230. The outer frame 240 extends downstream beyond the end cap 250 and accommodates a part of the tobacco capsule 30F. The end cap 250 is a cover that closes the gap between the channel forming body 230 and the outer frame 240 from the downstream side. The end cap 250 suppresses the aerosol source 200 stored in the cigarette storage 210 from leaking to the tobacco capsule 30F side.

[0256] [Combustion type aroma inhaler and its manufacturing method]

[0257] According to the method of the present embodiment, the manufacturing method of the combustion type flavor inhaler of the present embodiment includes: a process of separating and purifying neophytadiene from tobacco leaves to obtain a neophytadiene composition, a process of adding the above-mentioned neophytadiene composition to a substrate, and a process of manufacturing a combustion type flavor inhaler comprising the above-mentioned substrate. In addition, the combustion type flavor inhaler of the present embodiment includes the neophytadiene composition of the present embodiment. The manufacturing method of the combustion type flavor inhaler of the present embodiment obtains the neophytadiene composition by the method of the present embodiment, so that the neophytadiene can be efficiently separated and purified from tobacco leaves, and a combustion type flavor inhaler that can provide a good fragrance can be efficiently manufactured. In addition, since the combustion type flavor inhaler of the present embodiment includes the neophytadiene composition obtained by the method of the present embodiment, a good fragrance can be provided.

[0258] In the method for producing the combustion-type flavor inhaler of the present embodiment, the method for adding the neophytadiene composition to the base material can be performed in the same manner as the method for producing the non-combustion heating-type flavor inhaler described above.

[0259] An example of the combustion type flavor inhaler of this embodiment is shown in Fig.10 .like Fig.10 As shown, the combustion type flavor inhaler 400 includes: a tobacco-containing section 401, and a filter section 402 adjacent to the tobacco-containing section 401. The tobacco-containing section 401 includes: a tobacco filler 403 and a rolling paper 404 rolled around the tobacco filler 403, wherein the tobacco filler 403 includes a tobacco material containing the neophytadiene composition of the present embodiment. As described above, the rolling paper 404 may include the neophytadiene composition of the present embodiment. The filter section 402 is not particularly limited as long as it has the function of a general filter. For example, a tow made of synthetic fibers (sometimes also simply referred to as a "tow") may be used, and an article made of paper or other materials processed into a cylindrical shape may be used. The tobacco-containing section 401 and the filter section 402 are connected by a tipping paper component 405 wound on the tobacco-containing section 401 and the filter section 402. The tipping paper component 405 may have ventilation holes on a portion of its periphery. The number of ventilation holes may be one or more, for example, 10 to 40. When the number of ventilation holes is more than one, the ventilation holes may be arranged in a row in a ring shape at the outer periphery of the tipping paper member 405. The plurality of ventilation holes may be arranged at substantially constant intervals. By providing the ventilation holes, air is sucked into the filter section 402 from the ventilation holes during inhalation. By diluting the mainstream smoke with external air from the ventilation holes, a product with a desired tar value may be designed.

[0260] The user can enjoy the aroma of the cigarette by lighting the head end of the tobacco-containing section 401 and sucking the mouth end of the filter section 402. It should be noted that the number of the filter section 402 is not limited to one, for example, multiple filter sections with different functions can be connected and configured.

[0261] Example

[0262] Hereinafter, specific examples of the present embodiment will be described, but the present invention is not limited to these.

[0263] [Reference Example 1] (Study on tobacco leaf varieties)

[0264] Weigh 200 g of finely ground tobacco leaves of yellow or Burley tobacco seeds and put them into a closed stainless-steel container with a capacity of 2500 ml. Then, add 2000 ml of hexane (manufactured by FUJIFILM Wako Pure Chemical Industries, Ltd., for high-performance liquid chromatography) and extract for 3 hours while stirring in a closed state in a 40 °C water bath. After extraction, separate the hexane solution and the extraction residue using a stainless-steel sieve with a mesh size of 250 μm to obtain approximately 1500 ml of hexane solution. Prepare a 0.1 mass% sulfuric acid aqueous solution in advance, mix this 0.1 mass% sulfuric acid aqueous solution and the previously obtained hexane solution at a solution ratio (volume ratio) of sulfuric acid aqueous solution:hexane solution = 1:3, further add 200 g of table salt, and shake well in a separatory funnel to perform a liquid-liquid extraction operation. At this time, components represented by nicotine transfer to the lower-layer sulfuric acid aqueous solution, and the hydrophobic active components of the tobacco leaves transfer to the upper-layer organic layer. After standing still sufficiently, take out the organic layer, add 200 g of anhydrous sodium sulfate and stir to perform a dehydration operation within the organic layer. Remove insoluble matters by filtration through filter paper. Further, use a rotary evaporator (manufactured by BUCHI, Japan) to remove hexane under reduced pressure to obtain 5.16 g of dry solid (yield 2.58%) from yellow tobacco seeds and 5.4 g of dry solid (yield 2.7%) from Burley tobacco seeds. Dissolve each dry solid in a hexane solvent and perform GC / MS analysis on the obtained solution under the conditions shown below.

[0265] Gas chromatography-mass spectrometry (GC / MS)

[0266] Device: Agilent Technologies 7890A / 5975C GC / MSD

[0267] <GC conditions>

[0268] Column: HP-5MS (manufactured by Agilent Technologies)

[0269] Inner diameter 0.25 mm × length 30 m, film thickness 0.25 μm

[0270] Injection volume: 1 μl

[0271] Injection mode: Split (10:1)

[0272] Inlet temperature: 270 °C

[0273] Septum purge flow rate: 5 ml / min

[0274] Carrier gas: Helium (He)

[0275] Column flow rate: 1 ml / min (constant flow rate mode)

[0276] Oven temperature: 40 °C (3 min) - 4 °C / min - 280 °C (20 min)

[0277] Transfer line temperature: 280 °C

[0278]

[0279] Solvent waiting time: 4 min

[0280] Ionization method: Electron impact ionization method (EI method), 70 eV

[0281] Ion source temperature: 230 °C

[0282] Quadrupole temperature: 150 °C

[0283] Measurement mode: Scan

[0284] MS scan range: m / z 26 - 450

[0285] Threshold: 50

[0286] Sampling rate: 2

[0287] Based on the total ion chromatogram obtained by the above GC / MS analysis, when extracting a single ion (m / z = 68) contained in the MS chromatogram of phytol and comparing the peak area values of each peak, it was confirmed that the peak area value of phytol in the yellow variety was about 1.4 times that of the burley variety. Therefore, it can be seen that as tobacco leaves, both the yellow variety and the burley variety are useful, but from the perspective of higher phytol yield, the yellow variety is more preferred.

[0288] [Reference Example 2] (Study on extraction solvent)

[0289] Weighed 5 g of finely ground tobacco leaves and placed them in a screw tube as a 100 ml container. After adding 50 ml of an organic solvent (n - hexane, ethyl acetate, ethanol, or methanol) and mixing well, it was left standing at room temperature for one day and night. The mixed solution was filtered through filter paper. After adding a small amount of anhydrous sodium sulfate to the obtained extract for dehydration and then filtering through filter paper again, the organic solvent was removed under reduced pressure. The dried solid was dissolved in the same solvent as used in extraction so that the concentration of the obtained dried solid was 4% by mass, and this solution was subjected to GC / MS analysis under the conditions shown in Reference Example 1. The total ion chromatograms obtained by the GC / MS analysis based on the n - hexane, ethyl acetate, ethanol, and methanol solutions are as Figures 11 to 14 shown.

[0290] According to Figures 11 to 14It can be seen from the chromatogram that when each solvent is used for extraction, neophytadiene (retention time 38.9 minutes) is extracted from tobacco leaf fine powder. Furthermore, it was confirmed that when ethyl acetate, ethanol and methanol are used for extraction, peaks of unnecessary components (retention time before 38 minutes) are present in the purification of neophytadiene. It can be seen that among the above four solvents, hexane, which does not contain the peaks of the above unnecessary components as much as possible, is more suitable as an extraction solvent.

[0291] [Example 1]

[0292] (Process A1)

[0293] 200 g of yellow tobacco leaf powder was weighed and placed in a 2500 ml sealed stainless steel container. Then, 2000 ml of hexane (Fuji Film Wako Pure Chemicals, for high performance liquid chromatography) was added and extracted for 3 hours in a 40°C warm bath while being sealed and stirred. After extraction, the hexane solution and the extraction residue were separated using a stainless steel sieve with a mesh size of 250 μm to obtain about 1500 ml of hexane solution.

[0294] (Process B1)

[0295] A 0.1% by mass aqueous sulfuric acid solution was prepared in advance, and the 0.1% by mass aqueous sulfuric acid solution and the previously obtained hexane solution were mixed at a solution ratio (volume ratio) of aqueous sulfuric acid solution: hexane solution = 1:3 to obtain 500 ml of a mixed solution. Further, 200 g of common salt was added and the mixture was fully shaken in a separatory funnel to perform a liquid-liquid extraction operation. At this time, components represented by nicotine were transferred to the aqueous sulfuric acid solution in the lower layer, and the hydrophobic active ingredients of the tobacco leaves were transferred to the organic layer in the upper layer. After standing sufficiently, the organic layer was taken out, and about 200 g of anhydrous sodium sulfate was added and stirred to perform a dehydration operation in the organic layer. Insoluble matter was removed by filtration with filter paper. The obtained solution was subjected to GC / MS analysis under the conditions shown in Reference Example 1. The results are shown in Fig.15 .Depend on Fig.15 As shown in the chromatogram, it is known that alkaloids represented by nicotine (retention time 24.6 minutes) can be removed by the presence of neophytadiene (retention time 38.9 minutes) and the above-mentioned liquid-liquid extraction operation.

[0296] (Process C1)

[0297] Methanol was added to the organic layer (hexane layer) obtained above so that the mixing ratio was organic layer: methanol = 2:1 (volume ratio). After being fully mixed, the mixture was placed in a refrigerator (4 degrees) for one night to perform a liquid-liquid extraction operation. At this time, tobacco leaf resin components represented by CBT were transferred to the methanol solution in the lower layer, and neophytadiene, higher hydrocarbons, etc. were transferred to the hexane solution in the upper layer. The hexane solution and methanol solution obtained by the extraction operation were subjected to GC / MS analysis under the conditions shown in Reference Example 1. The results of the hexane solution and the results of the methanol solution are shown in Fig.16 , Fig.17 .according to Fig.16 , 17 The chromatogram shown in confirms that CBT is selectively removed by this step.

[0298] (Process D1, E1 and F1)

[0299] The hexane solution obtained above was removed under reduced pressure using a rotary evaporator (manufactured by BUCHI, Japan) to obtain 3.15 g of a dry solid (yield 1.58%). 300 mL of methanol was added to the dry solid, and the dry solid was dispersed and dissolved in methanol while being ultrasonically treated. The obtained solution was allowed to stand in a refrigerator (4°C) for 1 hour, and the supernatant from which the precipitate was removed was recovered. The supernatant was subjected to GC / MS analysis under the conditions shown in Reference Example 1. The results are shown in Fig.18 It was confirmed that higher hydrocarbons were selectively removed by this step.

[0300] (Process G1 and H1)

[0301] Methanol was removed from the supernatant obtained above under reduced pressure using a rotary evaporator (manufactured by BUCHI, Japan). Hexane (15 mL) was added to the obtained dry solid (1.49 g) to dissolve it. Neophytadiene was separated and purified from the obtained solution using normal phase HPLC. 1260 Infinity (trade name, manufactured by Agilent Technologies) was used as HPLC. The separation conditions of HPLC are as follows.

[0302] ·Chromatographic column: YMC-Pack SIL / S-5μm / 12nm manufactured by YMC Corporation

[0303] Mobile phase: Hex (2 min) → Hex:AcOEt (95:5) (1 min) → Hex (12 min)

[0304] Flow rate: 7mL / min

[0305] Detector: Diode array detector (220nm)

[0306] Sample injection volume: 500μL

[0307] After separation under the above conditions, the signal detected at 220 nm was separated in the fraction collector. The separated fractions are Fig.19 The fraction is shown in the dotted line. The fraction is recovered, and the obtained sample is concentrated and dried to obtain a colorless and transparent neophytadiene composition. According to the total ion chromatogram obtained by GC / MS analysis, the yield of neophytadiene is 65.0%, confirming that neophytadiene can be separated and purified from tobacco leaves with a high yield. In addition, the obtained neophytadiene composition was dissolved in methanol and GC / MS analysis was performed under the conditions shown in Reference Example 1. The results are shown in Fig. 20 When the purity was calculated, it was found that the purity of the neophytadiene composition was 99.83%, confirming that neophytadiene could be separated and purified at a high purity by the method of this embodiment.

[0308] [Example 2]

[0309] 1000 mass ppm (alcohol preparation) of the neophytadiene composition obtained in Example 1 was added to tobacco shreds for flavoring. The obtained flavored shreds were dried, filled into the tobacco rod portion of an externally heated non-combustion heating type flavor inhaler, and evaluated. The unflavored shreds were also filled into the tobacco rod portion of an externally heated non-combustion heating type flavor inhaler in the same manner and evaluated. As a result, the flavored shreds better reflected the original important flavor of tobacco than the unflavored shreds.

[0310] [Example 3]

[0311] 500 mass ppm (alcohol preparation) of the neophytadiene composition obtained in Example 1 was added to the tobacco shreds for flavoring. The obtained flavored tobacco shreds were dried, filled into the tobacco-containing section of the combustion-type flavor inhaler, and evaluated. The unflavored tobacco shreds were also filled into the tobacco-containing section of the combustion-type flavor inhaler in the same manner and evaluated. As a result, the flavored tobacco shreds better reflected the original important flavor of tobacco than the unflavored tobacco shreds.

Claims

1. A method for separating and purifying neophytadiene from tobacco leaves, the method comprising: A step of adding an aprotic solvent to the tobacco leaves for extraction, removing the extraction residue, and obtaining an extract (A1); A step of adding an acidic aqueous solution to the extract (A1) for liquid-liquid extraction, removing the aqueous layer, and obtaining 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 solid (D1); A step of adding a protic solvent to the dry solid (D1) to dissolve the dry solid (D1) to obtain a solution (E1); A step of removing the precipitate generated 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); and A step of separating and purifying neophytadiene from the solution (G1) by normal phase high performance liquid chromatography.

2. A method for separating and purifying neophytadiene from tobacco leaves, the method comprising: A step of adding an aprotic solvent to the tobacco leaves for extraction, removing the extraction residue, and obtaining an extract (A2); A step of adding an acidic aqueous solution to the extract (A2) for liquid-liquid extraction, removing the aqueous layer, and obtaining an extract (B2); A step of removing the solvent from the extract (B2) to obtain a dry solid (C2); A step of adding a protic solvent to the dry solid (C2) to dissolve the dry solid (C2) to obtain a solution (D2); A step of removing the precipitate generated 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, removing the protic solvent layer, and obtaining a solution (F2); as well as A step of separating and purifying neophytadiene from the solution (F2) by normal phase high performance liquid chromatography.

3. The method according to claim 1 or 2, wherein: The pH of the acidic aqueous solution is 3 or less.

4. The method according to any one of claims 1 to 3, wherein: The aprotic solvent is at least one solvent selected from hexane, heptane and ethyl acetate.

5. The method according to any one of claims 1 to 4, wherein: The protic solvent is methanol.

6. The method according to any one of claims 1 to 5, wherein: The tobacco leaf is at least one of yellow tobacco and burley tobacco.

7. 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 the protic solvent layer to obtain the solution (G1).

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

9. The method according to any one of claims 1 to 8, wherein: The neophytadiene composition obtained by separation and purification contains more than 95% by mass of neophytadiene.

10. A method for manufacturing a non-combustion heating type aroma inhaler, comprising: A step of separating and purifying neophytadiene from tobacco leaves by the method according to any one of claims 1 to 9 to obtain a neophytadiene composition; The step of adding the neophytadiene composition to a substrate; A process for manufacturing a non-combustion heating type flavor inhaler including the substrate.

11. A method for manufacturing a non-combustion and non-heating type aroma inhaler, comprising: A step of separating and purifying neophytadiene from tobacco leaves by the method according to any one of claims 1 to 9 to obtain a neophytadiene composition; The step of adding the neophytadiene composition to a substrate; and The invention relates to a process for manufacturing a non-combustion and non-heating type flavor inhaler including the substrate.

12. A method for manufacturing a combustion type aroma inhaler, comprising: A step of separating and purifying neophytadiene from tobacco leaves by the method according to any one of claims 1 to 9 to obtain a neophytadiene composition; The step of adding the neophytadiene composition to a substrate; and A process for manufacturing a combustion-type flavor inhaler including the substrate.

13. A neophytadiene composition comprising 95% by mass or more of neophytadiene, The composition is obtained by separating and purifying neophytadiene from tobacco leaves by the method according to any one of claims 1 to 9.

14. A non-combustion heating type flavor inhaler comprising the neophytadiene composition according to claim 13.

15. A non-combustion and non-heating type flavor inhaler, comprising the neophytadiene composition according to claim 13.

16. A combustion type flavor inhaler comprising the neophytadiene composition according to claim 13.

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