Extraction and purification method of sclareolide

Abstract

The invention provides an extraction and purification method of sclareolide. The extraction and purification method comprises the following steps: mixing a sclareolide raw material with an organic solvent, and collecting an organic phase to obtain a sclareolide mixed solution; concentrating the sclareolide mixed solution, and carrying out cooling crystallization to obtain a sclareolide coarse crystal; and carrying out recrystallization or crystal washing on the sclareolide coarse crystal to obtain the sclareolide, the sclareolide raw materials comprise the sclareolide and sclareol. The method can solve the problem of complex process for extracting and purifying sclareolide in the prior art, and is suitable for the field of preparation of sclareolide.

Description

Technical Field

[0001] The invention relates to the field of sclareolide preparation, and in particular to a method for extracting and purifying sclareolide. Background Art

[0002] Sclareolide is an important ambergris-flavored synthetic flavoring agent with extensive applications in cosmetics, tobacco, food, and other fields. In the cosmetics and perfume industry, it is primarily used in the synthesis of ambroxan, a natural ambergris substitute, and can also be used in the blending of flavors. In the tobacco industry, as an excellent flavor enhancer, it can mask tobacco's harsh odors, improve and enhance the flavor quality, imparting a captivating amber aroma and making cigarettes softer and more mellow. In the food industry, it can enhance and improve the sensory properties of food, serving as a flavoring agent to enhance the olfactory and taste effects of food. Sclareolide can also help improve weight loss and is therefore widely used in weight loss products. Currently, sclareolide is primarily used in the synthesis of ambergris substitute, ambroxan.

[0003] The synthesis methods of sclareolide are mainly divided into chemical synthesis and biotransformation. The former uses sclareol as a substrate through a series of redox reactions, such as traditional methods such as chromic anhydride oxidation or potassium permanganate oxidation. These methods have the advantages of short synthesis cycles and high yields, but the oxidants are expensive, post-processing is difficult, and they produce large amounts of heavy metal wastewater, seriously polluting the environment. Biotransformation, which uses microbial fermentation to convert sclareol into sclareolide, has the advantages of mild reaction conditions, good selectivity, and environmental friendliness. However, highly active strains are rare, the reaction time is long, and the yield is relatively low. Currently, chemical synthesis is the main method used in production, and reports on the biotransformation of sclareolide are relatively rare. However, considering the sustainable development and environmental friendliness, biotransformation may partially replace chemical synthesis in the future.

[0004] Currently, there are few reports on the extraction and purification methods of sclareolide, and the process maturity is low. In the existing technology, the extraction of sclareolide mostly uses equipment such as column chromatography or molecular distillation. The purification process is complex, the cost is high, and it is not easy to achieve in production, especially large-scale production. Summary of the Invention

[0005] The main purpose of the present invention is to provide a method for extracting and purifying sclareolide, so as to solve the problem of complex process for extracting and purifying sclareolide in the prior art.

[0006] In order to achieve the above-mentioned purpose, according to the first aspect of the present invention, a method for extracting and purifying sclareolide is provided, which comprises: mixing a sclareolide raw material with an organic solvent, collecting the organic phase, and obtaining a sclareolide mixed liquid; concentrating the sclareolide mixed liquid, cooling and crystallizing it, and obtaining crude sclareolide crystals; recrystallizing or washing the crude sclareolide crystals to obtain sclareolide; the sclareolide raw material comprises sclareolide and sclareol alcohol.

[0007] Furthermore, the organic solvent includes one or more of ethanol, ethyl acetate or isopropanol, the ethanol includes anhydrous ethanol or hydrous ethanol, and the volume percentage of ethanol in the hydrous ethanol is ≥95%; preferably, the sclareolactone raw material is mixed with the organic solvent, and the sclareolactone in the sclareolactone raw material is extracted, the extraction temperature is 40-60°C, and the extraction time is 1-5h; preferably, the number of extractions is 2-4 times, and the organic phases are combined to obtain a sclareolactone mixed solution; preferably, the mass volume ratio of the sclareolactone raw material to the organic solvent is 1kg:5-8L.

[0008] Furthermore, concentrating the sclareolactone mixed liquid includes: when the organic solvent is ethanol, concentrating the sclareolactone mixed liquid to a nearly saturated sclareolactone mixed liquid, and then cooling and crystallizing the nearly sclareolactone mixed liquid; when the organic solvent is not ethanol, concentrating the sclareolactone mixed liquid to a solid crude product free of organic solvent, mixing the solid crude product with ethanol to obtain a nearly saturated sclareolactone mixed liquid, and cooling and crystallizing the nearly sclareolactone mixed liquid; the concentration of sclareolactone in the nearly saturated sclareolactone mixed liquid is greater than 90% of the solubility of sclareolactone and less than the solubility of sclareolactone.

[0009] Furthermore, the temperature of the nearly sclareolide saturated mixed solution is 40-60°C.

[0010] Furthermore, concentration includes concentration by reduced pressure evaporation, concentration by normal pressure evaporation or concentration by membrane separation.

[0011] Furthermore, the temperature of the crystallization by cooling is 4-25°C; preferably 4-15°C; more preferably 7-10°C; preferably, the time of the crystallization by cooling is 12-48h.

[0012] Furthermore, the recrystallization includes: using anhydrous ethanol to recrystallize the crude crystals of sclareolide 1-2 times to obtain sclareolide.

[0013] Furthermore, the crystal washing comprises: washing the crude sclareolide crystals with aqueous ethanol to obtain sclareolide; preferably, the volume ratio of the crude sclareolide crystals to the aqueous ethanol is 1:2-1:4.

[0014] Furthermore, the volume percentage of ethanol in the hydrous ethanol is 55 to 75%.

[0015] Furthermore, the sclareolide raw material is a product of preparing sclareolide by biotransformation; preferably, the sclareolide raw material includes fermentation broth or dried fermentation product.

[0016] By applying the technical solution of the present invention, the prepared sclareolactone raw material is extracted and purified using the above-mentioned extraction and purification method through cooling crystallization, recrystallization or crystal washing, etc., which can stably prepare high-purity sclareolactone, overcoming the shortcomings of the more complicated extraction and purification processes of most lactone products. DETAILED DESCRIPTION

[0017] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present invention will be described in detail below with reference to the embodiments.

[0018] As mentioned in the background art, there are currently few reports on the extraction and purification methods for sclareolide, and the process maturity is low. In the prior art, column chromatography or molecular distillation are often used to extract such lactone products. For example, the molecular distillation device mentioned in CN1594303A "Method for separating and purifying sclareolide from sclareol reaction synthesis products" is used to purify sclareolide. It performs high-temperature and high-pressure distillation on the reaction compound and continuously separates it step by step from the residual distillation to obtain high-purity sclareolide. Although this method is simple and does not use organic solvents, the equipment cost is high and large-scale production is not easy to achieve. If purification is performed by chromatography, the extract needs to be subjected to multiple adsorption and elution using resin in CN116143799A, and the purification process is complicated, the cost is high, and it is not easy to achieve in production, especially large-scale production. Therefore, in the present application, the inventors attempt to develop a simple process for the extraction and purification of sclareolide and propose a series of protection schemes for the present application.

[0019] In a first typical embodiment of the present application, a method for extracting and purifying sclareolide is provided, which comprises: mixing a sclareolide raw material with an organic solvent, collecting an organic phase, and obtaining a sclareolide mixed solution; concentrating the sclareolide mixed solution, cooling and crystallizing the mixed solution, and obtaining crude sclareolide crystals; recrystallizing or washing the crude sclareolide crystals to obtain sclareolide; the sclareolide raw material comprises sclareolide and sclareol alcohol.

[0020] The above-mentioned sclareolide extraction and purification method can be used to extract and purify sclareolide from a sclareolide raw material. The above-mentioned sclareolide raw material includes, but is not limited to, sclareolide raw materials prepared by chemical synthesis or biotransformation. The above-mentioned sclareolide raw materials include both the target product sclareolide and the reaction raw material sclareolol, as well as other raw materials or intermediates. The above-mentioned sclareolide raw materials include dry powders obtained by drying and aqueous solutions obtained by biotransformation. The extraction method for the dry powder is to mix the dry powder with an organic solvent and obtain an organic phase by filtration. The extraction method for the aqueous solution is to extract the aqueous solution with an organic solvent and collect the organic phase by separation. In both cases, the sclareolide raw material is mixed with an organic solvent to extract a sclareolide mixed solution containing sclareolide and structural analogs such as sclareol from the sclareolide raw material.

[0021] After obtaining the mixed solution, the sclareolide mixed solution is concentrated and then cooled and crystallized. As the temperature decreases, the solubility of sclareolide decreases, and the concentrated sclareolide with a high concentration is precipitated and crystallized, thereby obtaining crude sclareolide crystals with high purity, thereby achieving a crude separation of sclareolide from other components such as sclareol or other impurities. The remaining mother liquor is a saturated solution of sclareolide, which can be further concentrated and crystallized in the future, or added to other batches of sclareolide mixed solution for concentration and subsequent cooling and crystallization, thereby increasing the yield of sclareolide.

[0022] The crude sclareolide crystals are then recrystallized or washed to remove impurities remaining on the crystal surface, further improving the purity of sclareolide, thereby obtaining sclareolide with higher purity.

[0023] In a preferred embodiment, the organic solvent includes one or more of ethanol, ethyl acetate or isopropanol, and the ethanol includes anhydrous ethanol or hydrous ethanol, and the volume percentage of ethanol in the hydrous ethanol is ≥95%; preferably, the sclareolactone raw material is mixed with the organic solvent, and the sclareolactone in the sclareolactone raw material is extracted, the extraction temperature is 40-60°C, and the extraction time is 1-5h; preferably, the number of extractions is 2-4 times, and the organic phases are combined to obtain a sclareolactone mixed solution; preferably, the mass volume ratio of the sclareolactone raw material to the organic solvent is 1kg:5-8L.

[0024] Sclareolide is extracted by mixing it with an organic solvent such as ethanol, ethyl acetate, or isopropanol. The extraction temperature is 40-60°C, including but not limited to 40, 45, 50, 55, or 60°C, and the extraction time is 1-5 hours. Compared to room temperature extraction, an extraction temperature of 40-60°C allows for better extraction of sclareolide, increases its solubility in the solution, reduces extraction time, and improves extraction yield. However, if the temperature is too high, other impurities may be introduced into the solution, and the organic solvent is prone to volatilization, which reduces the extraction solvent used, resulting in a lower extraction yield and hindering industrial production, posing a safety hazard. The mass-to-volume ratio of the sclareolide raw material to the organic solvent is 1 kg:5-8 L, including but not limited to 1 kg:5 L, 1 kg:5.5 L, 1 kg:6 L, 1 kg:6.5 L, 1 kg:7 L, 1 kg:7.5 L, or 1 kg:8 L. The amount of organic solvent mentioned above includes the total volume of solvent used in multiple extractions. Utilizing this ratio of raw material to organic solvent allows for efficient extraction of the target product, sclareolide, from the raw material, while also reducing solvent waste caused by excessive use and avoiding the excessive workload of the concentration process caused by the use of large amounts of solvent. The volume percentage of ethanol in the aqueous ethanol used for extraction is ≥ 95%, including but not limited to 95%, 96%, 97%, 98%, 99%, or 99.5%.

[0025] In a preferred embodiment, concentrating the sclareolactone mixed solution includes: when the organic solvent is ethanol, concentrating the sclareolactone mixed solution to a nearly saturated sclareolactone mixed solution, and then cooling and crystallizing the nearly saturated sclareolactone mixed solution; when the organic solvent is not ethanol, concentrating the sclareolactone mixed solution to a solid crude product without organic solvent, mixing the solid crude product with ethanol to obtain a nearly saturated sclareolactone mixed solution, and cooling and crystallizing the nearly saturated sclareolactone mixed solution; the concentration of sclareolactone in the nearly saturated sclareolactone mixed solution is greater than 90% of the solubility of sclareolactone and less than the solubility of sclareolactone.

[0026] For the sclareolide raw material of solid (including but not limited to dry powder) state, preferably ethanol is utilized to extract the sclareolide raw material, organic phase sclareolide mixture can be obtained by filtration, the sclareolide mixture obtained by lixiviate can directly carry out decrease temperature crystallization after concentrating, it is not necessary to reuse ethanol dissolution after the sclareolide mixture is dried to obtain solid crude product, it is possible to reduce the required step and solvent type for production, and reduce the impact of other solvents on purification effect. For the sclareolide raw material of liquid state, preferably ethyl acetate is utilized to mix, organic phase is obtained by extraction, it is possible to save the drying step for raw material. The nearly saturated mixed solution of the sclareolide obtained by above-mentioned concentration or dissolution method is a solution close to saturation of sclareolide, utilize this nearly saturated solution, it is possible to prevent that too high concentration directly separates out solid or part of solid can not dissolve before crystallization step, the solid purity produced by this part is lower, affects the purity of overall product.

[0027] The above-mentioned cooling crystallization preferably occurs in ethanol. In the crystallization carried out in ethanol, a large amount of sclareolide is precipitated, and sclareol and other impurities are almost not precipitated. The purity and yield of the target product are high, and sclareolide does not react with ethanol to generate other impurities.

[0028] In a preferred embodiment, the temperature of the nearly sclareolide mixed solution is 40-60°C, including but not limited to 40, 45, 50, 55 or 60°C.

[0029] The sclareolide mixture is concentrated to obtain a nearly sclareolide-saturated mixed solution, and the temperature is maintained at 40-60°C. The solubility of sclareolide increases with increasing temperature, so maintaining a higher temperature allows more sclareolide to be dissolved per unit volume of the mixed solution, thereby precipitating more crude sclareolide crystals in the subsequent cooling crystallization step, thereby improving the purification yield and efficiency.

[0030] The nearly saturated sclareolide mixture is preferably an ethanol solution. In ethanol, the solubility of sclareolide varies significantly at different temperatures, and its solubility is lower in ethanol at lower temperatures. Therefore, the crude sclareolide crystals precipitated by cooling are of high purity and are precipitated in large quantities. Furthermore, sclareolide does not react with ethanol, preventing the target product from reacting with the solvent, resulting in low yield and low purity.

[0031] In a preferred embodiment, the concentration includes concentration by reduced pressure evaporation, concentration by normal pressure evaporation or concentration by membrane separation.

[0032] Vacuum evaporation concentration is the concentration of a solution under reduced pressure (less than 1 standard atmosphere). Lowering the pressure lowers the boiling point of the solution, which can accelerate the evaporation of the solution. Atmospheric pressure evaporation concentration is the evaporation concentration of a solution at room pressure.

[0033] In a preferred embodiment, the temperature of the crystallization by cooling is 4-25°C; preferably 4-15°C; more preferably 7-10°C; preferably, the time of the crystallization by cooling is 12-48h.

[0034] In the above-mentioned crystallization by cooling, the temperature of the crystallization by cooling is 4-25°C, preferably 4-15°C, more preferably 7-10°C, including but not limited to 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25°C. Within the above-mentioned temperature range, the solubility of sclareolide is low, and sclareolide crystals with higher purity can be precipitated. The temperature of the crystallization by cooling should not be too low, otherwise the solubility of impurities such as sclareol will also be reduced, resulting in precipitation, affecting the purity of the target product; the temperature should not be too high, otherwise the yield of sclareolide will be reduced, and a large amount of sclareolide will be dissolved in the mother liquor and cannot be precipitated. The cooling crystallization time includes but is not limited to 12, 14, 16, 18, 20, 24, 28, 32, 36, 40 or 48 hours. If the crystallization time is too short, it will lead to insufficient crystallization of sclareolide, affecting the yield.

[0035] In a preferred embodiment, the recrystallization comprises: using anhydrous ethanol to recrystallize the crude sclareolide crystals 1-2 times to obtain sclareolide.

[0036] In a preferred embodiment, the crystal washing comprises: washing crude sclareolide crystals with aqueous ethanol to obtain sclareolide.

[0037] In a preferred embodiment, the volume ratio of the crude sclareolide crystals to the aqueous ethanol is 1:2-1:4.

[0038] In a preferred embodiment, the volume percentage of ethanol in the hydrous ethanol is 55-75%.

[0039] The aqueous ethanol used for washing the crystals has a different volume percentage of ethanol from that used for extraction. The crude sclareolide crystals are washed with aqueous ethanol, wherein the volume percentage of ethanol in the aqueous ethanol is 55-75%, including but not limited to 55%, 60%, 65%, 70%, or 75%. Compared to anhydrous ethanol, aqueous ethanol with such a composition has a higher polarity and can remove sclareol and other impurities with higher polarity remaining on the surface of the crude crystals, thereby reducing the dissolution of sclareolide, thereby improving the purity of the target product while ensuring the yield of the target product.

[0040] In a preferred embodiment, the sclareolide raw material is a product produced by biotransformation; preferably, the sclareolide raw material includes a fermentation broth or a dried fermentation product.

[0041] The aforementioned biotransformation products for preparing sclareolide, i.e., sclareolide raw materials, include but are not limited to products produced by the microbial or enzymatic conversion of sclareol or other compounds to sclareolide. Since the reactions in these raw materials mostly occur in an aqueous phase, free from residual organic solvents or oxides, they are more conducive to obtaining a higher-purity sclareolide product, with sclareolide having a purity exceeding 97% (vapor phase purity) being possible.

[0042] The beneficial effects of the present application will be further explained in detail below with reference to specific embodiments.

[0043] Example 1

[0044] 1 kg of dry powder containing sclarea lactone was put into a reactor, and 3 L of ethyl acetate was added thereto. The mixture was stirred and extracted at 25 ° C for 2 h. The filtrate was collected by filtration using a Buchner funnel. The separated heavy phase was continued to be put into the reactor and 2 L of ethyl acetate was added for a second extraction. After separation, 2 L of ethyl acetate was continued to be added for a third extraction. After filtration, 5.6 L of the filtrate was combined (gas phase detection showed that the product contained 99.6 g).

[0045] Pour 5.6 L of the separated filtrate (containing 99.6 g of product) into a rotary evaporator and concentrate under reduced pressure at 60°C until the solvent is essentially gone and all the solid crude product remains. Add 0.5 L of anhydrous ethanol to the crude solid product and dissolve it completely at 50°C before pouring it into a beaker. Place the solution in a 10°C room for cooling and crystallization. After 24 hours of crystallization, filter and separate, collecting 69.7 g of crystals.

[0046] About 210 mL of 70% ethanol (crystal mass: 70% ethanol volume = 1:3) was added to the separated crystals, and the crystals were washed with stirring. After 10 minutes, the crystals were separated and collected by filtration and dried to obtain 67.2 g of clarylide crystals (gas phase detection purity was 97.5%).

[0047] The mother liquor after the crystallization was concentrated again, and then cooled (at 10° C.) for crystallization. After subsequent separation, washing and drying, 23.3 g of sclareolide (purity 95.4%) was obtained. The yield of Example 1 was 88.21% based on the total product amount.

[0048] Example 2

[0049] 1 kg of dry powder containing sclarea lactone was put into a reactor, and 3 L of anhydrous ethanol was added thereto. The mixture was stirred and extracted in a 50°C water bath for 2 h. The filtrate was collected by filtration using a Buchner funnel. The separated heavy phase was then put into the reactor and 2 L of anhydrous ethanol was added for a second extraction. After separation, 2 L of anhydrous ethanol was added for a third extraction. After filtration, 5.4 L of the filtrate was combined (gas phase detection contained 108 g of product).

[0050] The separated filtrate (5.4 L) (containing 108 g of product) was poured into a rotary evaporator and concentrated under reduced pressure at 50°C until 0.5 L of solution remained. The concentrate was poured into a beaker. The concentrate was placed in a 4°C refrigerator for cooling and crystallization. After crystallization for 24 hours, it was separated by suction and separated, and 77.2 g of crystals were collected.

[0051] About 300 mL of 70% ethanol was added to the separated crystals, and the mixture was stirred to wash the crystals. After 10 min, the crystals were separated and collected by filtration and dried to obtain 63.1 g of sclareolide crystals (purity determined by gas chromatography was 97.3%).

[0052] The mother liquor after the crystallization was concentrated again, then cooled (at 4° C.) for crystallization for 24 hours, and then separated, washed, and dried to obtain 35.4 g of sclareolide (purity 96.1%). The yield of Example 2 was calculated to be 88.33% based on the total product amount.

[0053] Compared with Example 1, Example 2 reduces the types of solvents and uses only one organic solvent, ethanol, to complete the extraction and purification of the target product, and reduces the process step of dissolving the crude product, making the process simpler.

[0054] Example 3

[0055] The experimental scheme is the same as that of Example 2, except that the temperature of the two cooling crystallizations is 25°C, the time of the two cooling crystallizations is 48 h, and 60.8 g of sclareolide crystals (purity by gas phase detection is 97.7%) are obtained in the first cooling crystallization, and 34.5 g of sclareolide crystals (purity by gas phase detection is 96.5%) are obtained in the mother liquor crystallization. The total product yield is 85.83%.

[0056] Example 4

[0057] The experimental scheme is the same as that of Example 2, except that the temperature of the two cooling crystallizations is 10°C, the time of the two cooling crystallizations is 16 h, and 62.8 g of sclareolide crystals (purity by gas phase detection is 97.6%) are obtained in the first cooling crystallization, and 35.7 g of sclareolide crystals (purity by gas phase detection is 96.3%) are obtained in the mother liquor crystallization. The total product yield is 88.58%.

[0058] Example 5

[0059] The experimental scheme is the same as that of Example 2, except that the temperature of the two cooling crystallizations is 7°C, the time of the two cooling crystallizations is 12 h, and 63.0 g of sclareolide crystals (purity by gas phase detection is 97.7%) are obtained in the first cooling crystallization, and 35.1 g of sclareolide crystals (purity by gas phase detection is 96.1%) are obtained in the mother liquor crystallization. The total product yield is 88.22%.

[0060] Example 6

[0061] The experimental scheme was the same as that of Example 2, except that 95% vol ethanol was used for stirring extraction. The first cooling crystallization obtained 62.3 g of sclareolide crystals (purity by gas phase detection was 97.2%), and the mother liquor crystallization obtained 35.0 g of sclareolide crystals (purity by gas phase detection was 96.0%). The total product yield was 87.19%.

[0062] Example 7

[0063] The experimental protocol was the same as that of Example 2, except that extraction was performed in a 40°C water bath for 5 h, with a total of four extractions. The amount of solvent used in the first three extractions was the same as in Example 2, and the amount added in the fourth extraction was 2 L. The first crystallization by cooling yielded 63.7 g of sclareolide crystals (purity by gas chromatography assay, 97.3%), and crystallization of the mother liquor yielded 35.1 g of sclareolide crystals (purity by gas chromatography assay, 96.0%), for a total product yield of 88.59%.

[0064] Example 8

[0065] The experimental protocol was the same as that of Example 2, except that extraction was performed in a 60°C water bath for 1 hour, with two extractions performed, the first using 4 L of anhydrous ethanol and the second using 3 L of anhydrous ethanol. The first crystallization by cooling yielded 62.1 g of sclareolide crystals (purity by gas chromatography assay: 97.2%), and the mother liquor crystallization yielded 35.4 g of sclareolide crystals (purity by gas chromatography assay: 96.0%), for a total product yield of 87.35%.

[0066] Example 9

[0067] The experimental protocol was the same as that of Example 2, except that the extraction was performed at room temperature (25°C) for 6 h, a total of one extraction, and 4 L of anhydrous ethanol. The first crystallization by cooling yielded 58.8 g of sclareolide crystals (purity by gas chromatography assay: 97.3%), and the mother liquor crystallization yielded 30.7 g of sclareolide crystals (purity by gas chromatography assay: 96.2%), for a total product yield of 80.31%.

[0068] Example 10

[0069] The experimental protocol was identical to that of Example 2, except that 1 kg of dry powder containing sclareolide was placed in a reactor, 3 L of anhydrous ethanol was added, and extraction was carried out in a 50°C water bath for 2 h with stirring. The filtrate was collected by filtration using a Buchner funnel. The separated heavy phase was then placed in a reactor and extracted a second time with 2 L of anhydrous ethanol. The filtrates were then combined after filtration. The final yield of the total product was 83.27%.

[0070] Example 11

[0071] The experimental protocol was identical to that of Example 1, except that 1 kg of dry powder containing sclareolide was placed in a reactor, 3 L of isopropanol was added, and extraction was carried out at 25°C for 2 h. The filtrate was separated and collected by suction filtration using a Buchner funnel. The separated heavy phase was then placed in the reactor and extracted a second time with 3 L of isopropanol. After separation, 2 L of isopropanol was added for a third extraction, and the mixture was filtered and combined. The final yield of the total product was 87.66%.

[0072] From the above description, it can be seen that the above-mentioned embodiments of the present invention achieve the following technical effects: the prepared sclareolactone raw material is extracted and purified by the above-mentioned extraction and purification method, and through the exploration of various experimental parameters, a simple extraction and purification method is obtained that can stably prepare high-purity sclareolactone, overcoming the shortcomings of the complex extraction and purification process of sclareolactone and the large amount and variety of organic solvents used.

[0073] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. A method for extracting and purifying sclareolide, characterized in that: The extraction and purification method comprises: mixing the sclareolide raw material with an organic solvent, collecting the organic phase, and obtaining a sclareolide mixed solution; Concentrating the sclareolide mixed solution and performing cooling crystallization to obtain sclareolide crude crystals; Recrystallizing or washing the crude sclareolide crystals to obtain the sclareolide; The sclareolide raw material includes sclareolide and sclareol.

2. The extraction and purification method according to claim 1, wherein The organic solvent includes one or more of ethanol, ethyl acetate or isopropanol, The ethanol includes anhydrous ethanol or hydrous ethanol, and the volume percentage of ethanol in the hydrous ethanol is ≥95%; Preferably, the sclareolide raw material is mixed with the organic solvent, and the sclareolide in the sclareolide raw material is extracted, the extraction temperature is 40-60° C., and the extraction time is 1-5 hours; Preferably, the extraction is performed 2-4 times, and the organic phases are combined to obtain the sclareolide mixed solution; Preferably, the mass volume ratio of the sclareolide raw material to the organic solvent is 1 kg:5-8 L.

3. The extraction and purification method according to claim 2, wherein Concentrating the sclareolide mixed solution comprises: When the organic solvent is ethanol, concentrating the sclareolide mixed solution to a sclareolide nearly saturated mixed solution, and then performing the cooling crystallization on the sclareolide nearly saturated mixed solution; When the organic solvent is not ethanol, concentrating the sclareolide mixed solution to obtain a solid crude product free of the organic solvent, mixing the solid crude product with the ethanol to obtain the sclareolide nearly saturated mixed solution, and performing the cooling crystallization on the sclareolide nearly saturated mixed solution; The concentration of the sclareolactone in the nearly sclareolactone mixed solution is greater than 90% of the solubility of the sclareolactone and less than the solubility of the sclareolactone.

4. The extraction and purification method according to claim 3, wherein The temperature of the nearly sclareolide mixed solution is 40-60°C.

5. The extraction and purification method according to claim 1, wherein The concentration includes concentration by reduced pressure evaporation, concentration by normal pressure evaporation or concentration by membrane separation.

6. The extraction and purification method according to claim 1, wherein The temperature of the cooling crystallization is 4-25°C; Preferably 4-15°C; more preferably 7-10°C; Preferably, the cooling crystallization time is 12-48 hours.

7. The extraction and purification method according to claim 1, wherein The recrystallization comprises: using anhydrous ethanol to recrystallize the crude sclareolide crystals 1-2 times to obtain the sclareolide.

8. The extraction and purification method according to claim 1, wherein The crystal washing comprises: washing the crude sclareolide crystals with aqueous ethanol to obtain the sclareolide; Preferably, the volume ratio of the mass of the crude sclareolide crystals to the aqueous ethanol is 1:2-1:

4.

9. The extraction and purification method according to claim 8, characterized in that The volume percentage of ethanol in the hydrous ethanol is 55-75%.

10. The extraction and purification method according to claim 1, characterized in that: The sclareolide raw material is a product prepared by biotransformation. Preferably, the sclareolide raw material includes fermentation broth or dried fermentation product.