Method for efficiently preparing hawthorn extract by high-voltage discharge and aqueous two-phase system

Abstract

The invention discloses a method for efficiently preparing hawthorn extract through high-voltage discharge and an aqueous two-phase system, and relates to the technical field of tobacco flavor extract preparation. According to the aqueous two-phase extraction method, two insoluble polymers or polymers and an inorganic salt solution are adopted, extraction is carried out according to the difference of partition coefficients of solutes in two phases, and the method has the advantages of being high in extraction efficiency, simple and easy to operate, mild in extraction condition, beneficial to keeping biological activity, environmentally friendly and the like; high-voltage discharge and an aqueous two-phase system are combined to extract the hawthorn extract, the yield and quality of the extract are improved, volatilization of heat-sensitive aroma components is reduced, and the extracted hawthorn extract has the advantages of increasing the aroma concentration of cigarettes, increasing the sweet feeling of the cigarettes, reducing the irritation of the cigarettes, improving the comfort of the cigarettes and the like.

Description

Technical Field

[0001] This invention relates to the field of tobacco flavoring extract preparation technology, and in particular to a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system. Background Technology

[0002] Hawthorn, also known as mountain hawthorn or mountain red hawthorn, is a plant of the Rosaceae family. It has a long history of being used for food and medicine in China. Hawthorn is widely cultivated and abundant in my country. It contains rich vitamins, organic acids, flavonoids and various trace elements, and has high nutritional and medicinal value. Its fruit is sweet and sour, which can help digestion, strengthen the stomach and quench thirst. At the same time, it also has the effects of promoting blood circulation, removing blood stasis, lowering blood lipids, regulating blood pressure, inhibiting tumors, anti-oxidation and enhancing human immunity.

[0003] In the field of tobacco production technology, hawthorn is mainly used to prepare extracts for flavoring cigarettes. Hawthorn extract is a brownish-yellow liquid with the sour, sweet, and fragrant characteristics of hawthorn fruit. It is easily soluble in ethanol and ether and is widely used in the food, beverage, alcohol, health product, daily chemical, and tobacco industries. Hawthorn extract is a traditional tobacco flavoring raw material that can effectively regulate the acidity and alkalinity of smoke, mask tobacco impurities, reduce spiciness and irritation, moderate the taste, enhance the aroma, and improve the taste and aroma quality of cigarettes. Therefore, it has been widely adopted by domestic cigarette companies.

[0004] During the research, it was found that domestic fragrance manufacturers use the traditional low-alcohol continuous extraction method to prepare hawthorn extract. However, because some aroma components have low solubility in alcohol, the extraction efficiency of latent aroma substances is low, which limits the application of hawthorn extract in cigarette flavoring.

[0005] High-voltage discharge is an emerging unconventional green extraction technology based on electro-hydraulic discharge. It induces chemical and physical reactions in water. When electron avalanches, known as "streamers," diffuse from the positive electrode to the negative electrode, electrical breakdown occurs in the water. Energy is injected directly into the aqueous solution through a plasma channel formed by the high-voltage discharge between the two submerged electrodes. The direct effect of electrical breakdown in the liquid damages cell structures and forms membrane pores. However, various phenomena occur during this process, including the generation of low-temperature plasma, the emission of high-intensity ultraviolet light, the generation of strong shock waves and cavitation bubbles, and the generation of free radicals from water photodissociation. The increased shock waves and cavitation bubble explosions lead to reduced particle size, cell membrane fragmentation, increased mass transfer, and increased release of intracellular compounds into the solvent. This extraction technology can significantly improve extraction efficiency, reduce solvent and energy consumption, and is environmentally friendly. Summary of the Invention

[0006] The purpose of this invention is to provide a method for the efficient preparation of hawthorn extract using high-voltage discharge and a two-phase aqueous system, in order to solve the problem in the above-mentioned background art that the traditional low-alcohol continuous extraction method for preparing hawthorn extract has low extraction efficiency due to the low solubility of some aroma components in alcohol.

[0007] To achieve the above objectives, the present invention provides the following technical solution: a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, comprising the following steps: S1. Dry the hawthorn, then grind the dried hawthorn into powder to make hawthorn powder, and sieve it for later use. S2. Take a certain amount of hawthorn powder, add distilled water, extract in water, and then filter to obtain a first filter residue and a first extract. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Pump the mixed solution to a high-voltage discharge treatment chamber for discharge treatment and filtration to obtain a secondary extract. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure, then slowly add anhydrous ethanol while stirring rapidly. After completion, place it in a refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. S5. Place the centrifuged liquid on a rotary evaporator and concentrate under reduced pressure to obtain hawthorn extract.

[0008] In a preferred embodiment of the present invention, in step S1, the hawthorn is dried at 80°C for 12 hours, and the sieve is a 60-mesh sieve.

[0009] In a preferred embodiment of the present invention, in step S2, the amount of distilled water added is 4 times the mass of hawthorn powder, and the hawthorn powder is extracted in water at 90°C for 2 hours.

[0010] In a preferred embodiment of the present invention, in step S3, the mixed solution is delivered to the high-voltage discharge treatment chamber by a peristaltic pump at a flow rate of 8-12 mL / min, and the discharge treatment time is 20-40 min.

[0011] In a preferred embodiment of the present invention, in step S3, the ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.24~0.28 g / mL, and the ethanol volume fraction is 40~60%.

[0012] In a preferred embodiment of the present invention, in step S3, the mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 0.8~1.2g:20~40mL.

[0013] In a preferred embodiment of the present invention, in step S3, a high-voltage pulse generator is installed in the high-voltage discharge processing chamber to realize high-voltage discharge technology. The electrode spacing of the high-voltage pulse generator is 2~6mm, the pulse width is 20μs, the pulse voltage is 9kV, and the pulse frequency is 100Hz.

[0014] In a preferred embodiment of the present invention, in step S4, the third extract is concentrated to 100 mL under reduced pressure, the amount of anhydrous ethanol added is 300 mL, the ethanol concentration is adjusted to 80%, and the temperature of the third extract in the refrigerator is 4°C.

[0015] In a preferred embodiment of the present invention, in step S5, the centrifuged liquid is concentrated under reduced pressure at 55°C and 0.09 MPa on a rotary evaporator.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: The aqueous two-phase extraction method of this invention consists of two immiscible polymers or a polymer and an inorganic salt solution. Extraction is performed by utilizing the difference in the partition coefficients of the solute between the two phases. This method features high extraction efficiency, simple and easy operation, mild extraction conditions, preservation of biological activity, and environmental friendliness. Combining high-voltage discharge with an aqueous two-phase system for extracting hawthorn extract improves the yield and quality of the extract and reduces the volatilization of heat-sensitive aroma components. The extracted hawthorn extract has advantages such as increasing the aroma concentration of cigarettes, enhancing the sweetness of cigarettes, reducing cigarette irritation, and improving cigarette comfort. Attached Figure Description

[0017] Figure 1 This is the total ion chromatogram of the hot reflux hawthorn extract of the present invention; Figure 2 This is the total ion current chromatogram of Example 3 of the present invention. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] This invention provides a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, comprising the following steps: S1. Dry the hawthorn to promote the Maillard reaction of proteins and sugars in the hawthorn at high temperature. Crush the dried hawthorn into powder to make hawthorn powder, and sieve it for later use. In step S1, the hawthorn is dried at 80°C for 12 hours, and the sieve is a 60-mesh sieve. S2. Take a certain amount of hawthorn powder, add distilled water, extract in water, and then filter to obtain a first filter residue and a first extract. In step S2, the amount of distilled water added is 4 times the mass of hawthorn powder, and the hawthorn powder is extracted in water at 90°C for 2 hours. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Pump the mixed solution to a high-voltage discharge treatment chamber for discharge treatment and filtration to obtain a secondary extract. In step S3, the mixed solution is delivered to the high-voltage discharge treatment chamber by a peristaltic pump at a flow rate of 8-12 mL / min, and the discharge treatment time is 20-40 min. The ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.24~0.28 g / mL, and the ethanol volume fraction is 40~60%. The mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 0.8~1.2g:20~40mL; A high-voltage pulse generator is installed in the high-voltage discharge processing chamber. Specifically, the high-voltage pulse generator is a TP3020 model. High-voltage discharge technology is achieved through the high-voltage pulse generator. The electrode spacing of the high-voltage pulse generator is 2~6mm, the pulse width is 20μs, the pulse voltage is 9kV, and the pulse frequency is 100Hz. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure, then slowly add anhydrous ethanol while stirring rapidly. After completion, place it in a refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. In step S4, the third extract is concentrated under reduced pressure to 100 mL, the amount of anhydrous ethanol added is 300 mL, the ethanol concentration is adjusted to 80%, and the temperature of the third extract in the refrigerator is 4°C. S5. The centrifuged liquid was concentrated under reduced pressure in a rotary evaporator to obtain hawthorn extract; In step S5, the centrifuged liquid is placed on a rotary evaporator and concentrated under reduced pressure at 55°C and 0.09 MPa.

[0020] Example 1

[0021] The present invention discloses a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, comprising the following steps: S1. Dry the hawthorn at 80℃ for 12 hours, then pulverize the dried hawthorn into powder to make hawthorn powder, and pass it through a 60-mesh sieve for later use. S2. Take a certain amount of hawthorn powder, add 4 times the weight of hawthorn powder in distilled water, soak in water at 90℃ for 2 hours, then filter to obtain a first filter residue and a first extract. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Send the mixed solution to the high-voltage discharge treatment chamber through a peristaltic pump at a flow rate of 8 mL / min, perform discharge treatment for 20 min and filter to obtain a secondary extract. In step S3, the ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.24 g / mL, and the ethanol volume fraction is 40%. The mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 0.8 g: 20 mL; The high-voltage pulse generator is equipped with an electrode spacing of 2mm, a pulse width of 20μs, a pulse voltage of 9kV, and a pulse frequency of 100Hz. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure to 100 mL, then slowly add 300 mL of anhydrous ethanol while stirring rapidly. Adjust the ethanol concentration to 80%. After completion, place it in a 4°C refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. S5. The centrifuged liquid was concentrated under reduced pressure at 55℃ and 0.09 MPa in a rotary evaporator to obtain hawthorn extract.

[0022] Example 2

[0023] The present invention discloses a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, comprising the following steps: S1. Dry the hawthorn at 80℃ for 12 hours, then pulverize the dried hawthorn into powder to make hawthorn powder, and pass it through a 60-mesh sieve for later use. S2. Take a certain amount of hawthorn powder, add 4 times the weight of hawthorn powder in distilled water, soak in water at 90℃ for 2 hours, then filter to obtain a first filter residue and a first extract. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Send the mixed solution to the high-voltage discharge treatment chamber through a peristaltic pump at a flow rate of 9 mL / min, perform discharge treatment for 25 min and filter to obtain a secondary extract. In step S3, the ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.25 g / mL, and the ethanol volume fraction is 45%. The mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 0.9 g: 25 mL; The high-voltage pulse generator is equipped with an electrode spacing of 3mm, a pulse width of 20μs, a pulse voltage of 9kV, and a pulse frequency of 100Hz. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure to 100 mL, then slowly add 300 mL of anhydrous ethanol while stirring rapidly. Adjust the ethanol concentration to 80%. After completion, place it in a 4°C refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. S5. The centrifuged liquid was concentrated under reduced pressure at 55℃ and 0.09 MPa in a rotary evaporator to obtain hawthorn extract.

[0024] Example 3

[0025] The present invention discloses a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, comprising the following steps: S1. Dry the hawthorn at 80℃ for 12 hours, then pulverize the dried hawthorn into powder to make hawthorn powder, and pass it through a 60-mesh sieve for later use. S2. Take a certain amount of hawthorn powder, add 4 times the weight of hawthorn powder in distilled water, soak in water at 90℃ for 2 hours, then filter to obtain a first filter residue and a first extract. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Send the mixed solution to the high-voltage discharge treatment chamber through a peristaltic pump at a flow rate of 10 mL / min, perform discharge treatment for 30 min and filter to obtain a secondary extract. In step S3, the ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.26 g / mL, and the ethanol volume fraction is 50%. The mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 1g:30mL; The high-voltage pulse generator is equipped with an electrode spacing of 4mm, a pulse width of 20μs, a pulse voltage of 9kV, and a pulse frequency of 100Hz. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure to 100 mL, then slowly add 300 mL of anhydrous ethanol while stirring rapidly. Adjust the ethanol concentration to 80%. After completion, place it in a 4°C refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. S5. The centrifuged liquid was concentrated under reduced pressure at 55℃ and 0.09 MPa in a rotary evaporator to obtain hawthorn extract.

[0026] Example 4

[0027] The present invention discloses a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, comprising the following steps: S1. Dry the hawthorn at 80℃ for 12 hours, then pulverize the dried hawthorn into powder to make hawthorn powder, and pass it through a 60-mesh sieve for later use. S2. Take a certain amount of hawthorn powder, add 4 times the weight of hawthorn powder in distilled water, soak in water at 90℃ for 2 hours, then filter to obtain a first filter residue and a first extract. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Send the mixed solution to the high-voltage discharge treatment chamber through a peristaltic pump at a flow rate of 11 mL / min, perform discharge treatment for 35 min and filter to obtain a secondary extract. In step S3, the ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.27 g / mL, and the ethanol volume fraction is 55%. The mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 1.1 g: 35 mL; The high-voltage pulse generator is equipped with an electrode spacing of 5mm, a pulse width of 20μs, a pulse voltage of 9kV, and a pulse frequency of 100Hz. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure to 100 mL, then slowly add 300 mL of anhydrous ethanol while stirring rapidly. Adjust the ethanol concentration to 80%. After completion, place it in a 4°C refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. S5. The centrifuged liquid was concentrated under reduced pressure at 55℃ and 0.09 MPa in a rotary evaporator to obtain hawthorn extract.

[0028] Example 5

[0029] The present invention discloses a method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, comprising the following steps: S1. Dry the hawthorn at 80℃ for 12 hours, then pulverize the dried hawthorn into powder to make hawthorn powder, and pass it through a 60-mesh sieve for later use. S2. Take a certain amount of hawthorn powder, add 4 times the weight of hawthorn powder in distilled water, soak in water at 90℃ for 2 hours, then filter to obtain a first filter residue and a first extract. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Send the mixed solution to the high-voltage discharge treatment chamber through a peristaltic pump at a flow rate of 12 mL / min, perform discharge treatment for 40 min and filter to obtain a secondary extract. In step S3, the ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.28 g / mL, and the ethanol volume fraction is 60%. The mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 1.2 g: 40 mL; The high-voltage pulse generator is equipped with an electrode spacing of 6mm, a pulse width of 20μs, a pulse voltage of 9kV, and a pulse frequency of 100Hz. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure to 100 mL, then slowly add 300 mL of anhydrous ethanol while stirring rapidly. Adjust the ethanol concentration to 80%. After completion, place it in a 4°C refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. S5. The centrifuged liquid was concentrated under reduced pressure at 55℃ and 0.09 MPa in a rotary evaporator to obtain hawthorn extract.

[0030] The hawthorn extracts obtained in Examples 1-5 were added to tobacco shreds to make cigarettes. Cigarettes were then evaluated by five provincial-level cigarette tasters using the usual evaluation methods in the cigarette industry. Each indicator was scored out of 10, with higher satisfaction scores receiving higher marks. The average score given by the five provincial-level tasters was calculated. A control group was prepared using hawthorn extract obtained by spraying the same mass of ethanol and by the hot reflux method. The evaluation results are shown in Table 1. Table 1. Results of the drug evaluation

[0031] In summary, the hawthorn extract obtained by high-voltage discharge combined with a two-phase aqueous system treatment has a higher content of heat-sensitive aroma substances in its volatile aroma components than the hawthorn extract obtained by the hot reflux method, and the types of aldehydes, ketones, esters, and other substances have all increased.

[0032] The total ion chromatogram of the hot reflux hawthorn extract, used as a control group, is shown below. Figure 1 As shown in Table 2, the X-axis represents retention time in minutes, and the Y-axis represents abundance. The GC-MS analysis results of the hot reflux hawthorn extract are presented. GC-MS represents gas chromatography-mass spectrometry. Table 2. GC-MS analysis results of hot reflux hawthorn extract

[0033]

[0034] In the examples of preparing hawthorn extract using high-voltage discharge combined with an aqueous two-phase system, the total ion chromatogram of Example 3, which showed the best flavoring effect, is as follows: Figure 2 As shown in Table 3, the X-axis represents retention time in minutes, and the Y-axis represents abundance. The GC-MS analysis results of hawthorn extract in Example 3 are shown in Table 3: Table 3 shows the GC-MS analysis results of hawthorn extract in Example 3.

[0035] Table 3. GC-MS analysis results of hawthorn extract in Example 3

[0036]

[0037] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system, characterized in that, Includes the following steps: S1. Dry the hawthorn, then grind the dried hawthorn into powder to make hawthorn powder, and sieve it for later use. S2. Take a certain amount of hawthorn powder, add distilled water, extract in water, and then filter to obtain a first filter residue and a first extract. S3. Add an ethanol-ammonium sulfate aqueous two-phase system solution to the primary filter residue to prepare a mixed solution. Pump the mixed solution to a high-voltage discharge treatment chamber for discharge treatment and filtration to obtain a secondary extract. S4. Mix the second extract and the first extract to obtain the third extract. Concentrate the third extract under reduced pressure, then slowly add anhydrous ethanol while stirring rapidly. After completion, place it in a refrigerator overnight and then centrifuge to recover the centrifuged liquid and precipitate. S5. Place the centrifuged liquid on a rotary evaporator and concentrate under reduced pressure to obtain hawthorn extract.

2. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 1, characterized in that: In step S1, the hawthorn is dried at 80°C for 12 hours, and the sieve is a 60-mesh sieve.

3. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 1, characterized in that: In step S2, the amount of distilled water added is 4 times the mass of hawthorn powder, and the hawthorn powder is extracted in water at 90°C for 2 hours.

4. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 1, characterized in that: In step S3, the mixed solution is delivered to the high-voltage discharge treatment chamber by a peristaltic pump at a flow rate of 8~12mL / min, and the discharge treatment time is 20~40min.

5. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 1, characterized in that: In step S3, the ammonium sulfate mass concentration in the ethanol-ammonium sulfate aqueous two-phase system solution is 0.24~0.28 g / mL, and the ethanol volume fraction is 40~60%.

6. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 5, characterized in that: In step S3, the mass ratio of the primary filter residue to the ethanol-ammonium sulfate aqueous two-phase system solution is 0.8~1.2g:20~40mL.

7. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 1, characterized in that: In step S3, a high-voltage pulse generator is installed in the high-voltage discharge treatment chamber to realize high-voltage discharge technology. The electrode spacing of the high-voltage pulse generator is 2~6mm, the pulse width is 20μs, the pulse voltage is 9kV, and the pulse frequency is 100Hz.

8. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 1, characterized in that: In step S4, the third extract is concentrated under reduced pressure to 100 mL, the amount of anhydrous ethanol added is 300 mL, the ethanol concentration is adjusted to 80%, and the temperature of the third extract in the refrigerator is 4°C.

9. The method for efficiently preparing hawthorn extract using high-voltage discharge and a two-phase aqueous system according to claim 1, characterized in that: In step S5, the centrifuged liquid is placed on a rotary evaporator and concentrated under reduced pressure at 55°C and 0.09 MPa.

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