Method for increasing content of geranone and furandiene in zedoary turmeric oil

Abstract

The invention provides a method for increasing the content of geranone and furandiene in zedoary turmeric oil, which comprises the following steps: activating secondary metabolism through moderate drought, low temperature or salt stress in a key growth period of zedoary turmeric, and promoting sesquiterpene accumulation; then, phosphorus-potassium fertilizer and zinc-boron trace elements are additionally applied, nitrogen fertilizer is reduced, and the component proportion is optimized; the preparation method comprises the following steps: washing, drying and crushing the rhizome of curcuma zedoary, adding water to infiltrate, extracting crude oil by water vapor, performing reduced pressure distillation and molecular distillation purification, finally filling nitrogen and sealing, and refrigerating at 2-4 DEG C and storing in a dark place. According to the method, the content and extraction rate of target components can be remarkably increased, purity is guaranteed, thermosensitive components are protected, pollution is reduced, and the storage period is prolonged.

Description

Technical Field

[0001] The invention relates to the technical field of zedoary turmeric oil extraction, and in particular to a method for increasing the contents of geranone and furanodiene in zedoary turmeric oil. Background Art

[0002] Curcuma oil, an important volatile oil extracted from the rhizome of Curcuma zedoaria, exhibits significant potential for applications in medicine, healthcare, and other fields. Its main active ingredients, geraniol and furanodienes, exhibit significant anti-tumor activity, inhibiting tumor cell proliferation and inducing apoptosis through multiple mechanisms. They also possess multiple pharmacological activities, including anti-inflammatory and antibacterial properties, making them important for the treatment and prevention of various diseases. However, the current field of zecuma oil extraction faces numerous challenges, making it difficult to meet the growing market demand for geraniol and furanodienes. Traditional methods of extracting zedoary turmeric oil, such as steam distillation and organic solvent extraction, have obvious limitations. Steam distillation is one of the more commonly used extraction methods and is also a prescribed method in the 2020 edition of the Chinese Pharmacopoeia. Its operating principle is based on the difference in boiling points between water and the components in zedoary turmeric oil. The zedoary turmeric oil is evaporated along with the water vapor by heating, and then separated by condensation. However, during the extraction process of this method, zedoary turmeric oil needs to be in a high-temperature environment for a long time. Heat-sensitive components such as geraniol and furanodiene are easily decomposed and isomerized due to high temperature, resulting in a significant reduction in their content, which seriously affects the quality and efficacy of zedoary turmeric oil. In addition, the extraction efficiency of steam distillation is relatively low, making it difficult to achieve efficient extraction of zedoary turmeric oil. Organic solvent extraction utilizes the principle of like dissolves like, using organic solvents to extract zedoary oil from the rhizomes of zedoary turmeric. While this method can improve the extraction rate of zedoary oil to a certain extent, the residual organic solvent problem has become a drawback that cannot be ignored. Residual organic solvents not only affect the purity of zedoary oil and reduce product quality, but may also cause potential harm to human health, limiting the application of zedoary oil in the fields of medicine and health products. In addition, the recovery and treatment of organic solvents during the organic solvent extraction process consumes a lot of energy and costs, increasing the production burden. In addition to extraction methods, the accumulation of active ingredients in Curcuma zedoaria during its growth is also constrained by a variety of factors. Environmental factors such as temperature, light, moisture, soil conditions, and cultivation practices all influence the synthesis and accumulation of geraniol and furanodienes in Curcuma zedoaria rhizomes. However, systematic research on the growth environment and cultivation management of Curcuma zedoaria remains insufficient, and a scientific and comprehensive regulatory framework has yet to be established, hindering the full potential of Curcuma zedoaria growth and the improvement of its active ingredient content. Although researchers have conducted extensive research in recent years on the extraction and cultivation of zedoary turmeric oil, attempting to employ novel technologies such as supercritical fluid extraction and microwave-assisted extraction, these technologies still face numerous challenges in practical application, including high equipment costs, complex process conditions, and difficulty in large-scale promotion. Furthermore, research on synergistically increasing the content of active ingredients in zedoary turmeric through environmental stress and nutritional regulation during its growth has been insufficiently thorough, hindering its effective application in the industrial production of zedoary turmeric oil. Therefore, developing a method that effectively increases the content of geraniol and furanodienes in zedoary turmeric oil, while also being highly efficient, environmentally friendly, and economical, has become a critical issue urgently needed to be addressed in the field of zedoary turmeric oil extraction technology. Summary of the Invention

[0003] The object of the present invention is to provide a method for increasing the content of geranone and furanodienes in zedoary turmeric oil. By regulating the growth process of zedoary turmeric and adopting a composite extraction and gradient purification process, the content of geranone and furanodienes in zedoary turmeric oil is effectively increased, thereby improving the quality of zedoary turmeric oil.

[0004] To achieve the above objectives, the present invention adopts the following technical means: A method for increasing the content of geranone and furadiene in zedoary turmeric oil comprises the following steps: Stress treatment: During the critical growth period of Curcuma zedoaria, moderate drought, low temperature or salt stress is used to activate the secondary metabolic pathway of Curcuma zedoaria and promote the accumulation of sesquiterpenes; Nutritional regulation: After stress treatment, increase the application of phosphorus, potassium fertilizers and trace elements such as zinc and boron, reduce nitrogen fertilizers, and optimize the ratio of active ingredients in the rhizomes; Curcuma zedoaria pretreatment: Wash and dry the rhizomes of Curcuma zedoaria to a moisture content of less than 10%, and grind them through a 40-60 mesh sieve to obtain Curcuma zedoaria powder, which provides a suitable raw material for subsequent extraction; Composite extraction process: Adding water to the zedoary turmeric powder in a ratio of 1:1.5 and soaking the mixture in a sealed environment for at least 18 hours; extracting crude zedoary turmeric oil by steam distillation of the soaked zedoary turmeric powder; The crude zedoary turmeric oil is first pretreated by vacuum distillation, and then subjected to two-stage distillation using molecular distillation technology under conditions of a vacuum degree of less than 10 Pa and a temperature of less than 100° C., i.e., the first stage separates low-boiling point impurities, and the second stage collects the target fraction to obtain high-purity zedoary turmeric oil; Stable storage: The purified zedoary oil is placed in a brown glass container, filled with nitrogen to replace the air, and then sealed. It is then stored in a refrigerator at 2-4°C away from light.

[0005] Preferably, the moderate drought stress in the adverse stress treatment is to control the soil moisture content at 40%-60% of the field holding capacity during the tuber swelling period, which lasts for 10-15 days.

[0006] Preferably, the low temperature stress in the adverse stress treatment is to control the ambient temperature at 10-15° C. for 7-10 days during the vigorous growth period.

[0007] Preferably, the salt stress in the adverse stress treatment is to apply a 0.2%-0.5% sodium chloride solution to the soil during the seedling stage, once every 10 days, for a total of 2-3 times.

[0008] Preferably, the nutritional regulation in the nutritional regulation is specifically as follows: applying 20-30 kg of superphosphate and 15-20 kg of potassium sulfate per mu, and spraying 0.1%-0.3% zinc sulfate solution and 0.05%-0.1% borax solution on the leaves once every 15 days, for a total of 3-4 times; the application amount of nitrogen fertilizer is controlled at 10-15 kg per mu.

[0009] Preferably, the specific method of the first stage separation in the gradient purification process is: open the 150 short-range molecular distillation system, adjust the system temperature to 50 ° C, the pressure to 4-10Kpa, add the crude zedoary oil to the feed port, adjust the flow rate to 2.5mL / min, the scraper speed to 220r / min, molecular distillation, and collect the light zedoary oil; The specific method for collecting the target fraction in the second stage is as follows: the distillation temperature of the molecular distillation system is adjusted to 45°C, the pressure is adjusted to 50Pa, the speed of the scraper is adjusted to 200r / min, and the flow rate is 2.0mL / min, the zedoary oil with light content is distilled in one stage, and the distillate is collected to obtain a zedoary oil product with high content of geraniol and furanodiene.

[0010] The present invention has the following beneficial effects: 1. Significantly Increased Active Ingredient Content: Stress treatment simulates the non-ideal growth environment of zedoary turmeric, stimulating its defense mechanisms and activating secondary metabolic pathways, leading to the synthesis of large quantities of sesquiterpenes. The accumulation of geraniol and furanodienes, key sesquiterpenes, has increased significantly. Subsequent nutritional regulation precisely provides the nutrients required for zedoary turmeric growth. The rational application of phosphorus and potassium fertilizers, as well as trace elements such as zinc and boron, provides the material basis for the synthesis of active ingredients, further optimizing the ratio of geraniol and furanodienes in the rhizome. Ultimately, the content of these two key components in the extracted zedoary turmeric oil far exceeds that of traditional methods.

[0011] 2. Efficient Extraction and High Purity Guarantee: The composite extraction process combines traditional steam extraction with molecular distillation. This process complies with the requirements of the 2020 edition of the Chinese Pharmacopoeia, Part I, and the resulting product meets national standards. The gradient purification process utilizes vacuum distillation and molecular distillation techniques to effectively remove low-boiling impurities and other interfering components, significantly improving the purity of zedoary turmeric oil and meeting the stringent requirements for high-quality raw materials in the pharmaceutical and high-end health product sectors.

[0012] 3. Effectively protect heat-sensitive components: The low-temperature environment of molecular distillation can minimize the decomposition and structural damage of heat-sensitive components such as geranone and furadiene, completely retain their biological activity, and ensure the efficacy of zedoary oil.

[0013] 4. Reduce environmental pollution and safety risks: This method does not add organic solvents, thus avoiding the potential harm to human health caused by organic solvent residues, improving the safety of zedoary oil products, and being more in line with the concepts of green chemistry and sustainable development.

[0014] 5. Extend the product storage period: Using brown glass containers to avoid light, filling with nitrogen to replace air, and low-temperature refrigeration storage methods can effectively isolate oxygen, light and other factors that cause the oxidation and degradation of zedoary oil, slow down the deterioration of its components, significantly improve the storage stability of zedoary oil, extend the shelf life of the product, and facilitate the transportation, storage and market circulation of the product. DETAILED DESCRIPTION

[0015] The technical solution of the present invention will be described clearly and completely below. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. Example 1

[0016] A method for increasing the content of geranone and furadiene in zedoary turmeric oil comprises the following steps: Adversity stress treatment: During the critical period of Curcuma zedoaria growth, moderate drought, low temperature or salt stress is used to activate the secondary metabolic pathway of Curcuma zedoaria and promote the accumulation of sesquiterpenes; among them, the specific methods of activating the secondary metabolic pathway of Curcuma zedoaria by moderate drought, low temperature and salt stress are: controlling the soil moisture content at 40%-60% of the field water holding capacity during the tuber swelling period for 10-15 days; controlling the ambient temperature at 10-15℃ during the vigorous growth period for 7-10 days; applying a sodium chloride solution with a concentration of 0.2%-0.5% to the soil during the seedling stage, once every 10 days, for a total of 2-3 times.

[0017] Nutritional regulation: After the stress treatment, increase the application of phosphorus, potassium fertilizers and trace elements of zinc and boron, reduce nitrogen fertilizers, and optimize the ratio of active ingredients in the rhizomes; among them, the specific method of increasing the application of phosphorus, potassium fertilizers and trace elements of zinc and boron and reducing nitrogen fertilizers is: apply 20-30kg of superphosphate and 15-20kg of potassium sulfate per mu, and spray 0.1%-0.3% zinc sulfate solution and 0.05%-0.1% borax solution on the leaves once every 15 days, for a total of 3-4 times; the application amount of nitrogen fertilizer is controlled at 10-15kg per mu.

[0018] Pretreatment of Curcuma zedoaria: Wash and dry the harvested Curcuma zedoaria rhizomes until the moisture content is less than 10%, and grind them through a 40-60 mesh sieve to obtain Curcuma zedoaria powder, which provides suitable raw materials for subsequent extraction.

[0019] Composite extraction process: adding water to the zedoary turmeric powder in a ratio of 1:1.5 and soaking the mixture in a closed environment for 18 hours; extracting crude zedoary turmeric oil from the soaked zedoary turmeric powder by steam distillation to obtain crude zedoary turmeric oil; The crude zedoary turmeric oil is first pretreated by vacuum distillation, and then subjected to two-stage distillation using molecular distillation technology under conditions of a vacuum degree of less than 10 Pa and a temperature of less than 100° C., i.e., the first stage separates low-boiling point impurities, and the second stage collects the target fraction to obtain high-purity zedoary turmeric oil. The specific methods of the first stage separation and the second stage collection are as follows: First stage separation: Open the 150 short-range molecular distillation system, adjust the system temperature to 50°C and the pressure to 4-10KPa, add the crude zedoary oil to the feed port, adjust the flow rate to 2.5mL / min, and the scraper speed to 220r / min, and perform molecular distillation to collect the light zedoary oil; Second-stage collection: The molecular distillation system temperature was adjusted to 45°C, the pressure was adjusted to 50 Pa, the scraper speed was adjusted to 200 r / min, and the flow rate was 2.0 mL / min. The light-removed zedoary oil was subjected to primary distillation and the distillate was collected to obtain a zedoaryaryl oil product with a high content of geraniol and furanodiene.

[0020] Stable storage: put the purified zedoary oil into a brown glass container, fill it with nitrogen to replace the air, seal it, and store it in a refrigerator at 2-4℃ away from light. Example 2

[0021] A method for increasing the content of geranone and furadiene in zedoary turmeric oil comprises the following steps: Stress treatment: At a zedoary turmeric cultivation base in Guangxi, well-grown and uniform zedoary turmeric plants were selected and subjected to moderate drought stress during the root expansion phase. The irrigation system was controlled to maintain soil moisture at 50% of field capacity for 12 days. During this period, soil moisture was regularly monitored to ensure it remained within the target range. Nutritional Control: Immediately after the drought stress treatment, perform nutritional control. Apply 25kg of superphosphate and 18kg of potassium sulfate per mu (approximately 1.5 acres) of fertilizer evenly around the roots of the zedoary plants, and shallowly till the soil to thoroughly mix the fertilizer. At the same time, spray the zedoary leaves with a 0.2% zinc sulfate solution and a 0.08% borax solution three times every 15 days. Each spraying should be performed in the evening on a clear, windless day to ensure the solution is evenly applied to the leaf surface. Curcuma zedoaria pretreatment: Harvest zedoaria after maturity. Excavate the rhizomes and clean them of dirt and impurities with clean water. Place them in a well-ventilated drying room and dry them using a combination of natural drying and hot air drying until the moisture content reaches 8%. The dried rhizomes are crushed in a grinder and passed through a 50-mesh sieve to obtain zedoaria powder. Composite extraction process: adding water to the zedoary turmeric powder in a ratio of 1:1.5 and soaking the mixture in a closed environment for 18 hours; extracting crude zedoary turmeric oil from the soaked zedoary turmeric powder by steam distillation to obtain crude zedoary turmeric oil; The crude zedoary turmeric oil is first pretreated by vacuum distillation, and then subjected to two-stage distillation using molecular distillation technology under conditions of a vacuum degree of less than 10 Pa and a temperature of less than 100° C., i.e., the first stage separates low-boiling point impurities, and the second stage collects the target fraction to obtain high-purity zedoary turmeric oil. The specific methods of the first stage separation and the second stage collection are as follows: First stage separation: Open the 150 short-range molecular distillation system, adjust the system temperature to 50°C and the pressure to 4-10KPa, add the crude zedoary oil to the feed port, adjust the flow rate to 2.5mL / min, and the scraper speed to 220r / min, and perform molecular distillation to collect the light zedoary oil; Second-stage collection: The distillation temperature of the molecular distillation system is adjusted to 45°C, the pressure is adjusted to 50 Pa, the speed of the scraper is adjusted to 200 r / min, and the flow rate is 2.0 mL / min. The zedoary oil with light content is distilled in the first stage and the distillate is collected to obtain a zedoary oil product with high content of geraniol and furanodiene.

[0022] Stable Storage: Place high-purity zedoary oil in a brown glass container and replace the air in the container with nitrogen to minimize oxygen levels. After sealing the container, store it in a refrigerator at 2-4°C away from light. Example 3

[0023] A method for increasing the content of geranone and furadiene in zedoary turmeric oil comprises the following steps: Stress treatment: Healthy zedoary zedoary seedlings were selected from a zedoary zedoary cultivation area in Yunnan Province and subjected to salt stress during the seedling stage. A 0.3% sodium chloride solution was evenly applied to the soil, once every 10 days for a total of two applications. Each application ensured that the solution evenly penetrated the soil, ensuring full contact with the zedoary zedoary roots. Nutritional regulation: After the salt stress treatment is completed, nutritional regulation is performed. 30 kg of superphosphate and 20 kg of potassium sulfate are applied per mu, using the same fertilization method as in Example 2. At the same time, 0.3% zinc sulfate solution and 0.1% borax solution are sprayed on the leaves once every 15 days for a total of 4 sprays. Pretreatment of Curcuma zedoaria: When the Curcuma zedoaria grows to maturity, harvest the rhizomes, wash them, dry them to a moisture content of 9%, and grind them through a 60-mesh sieve to obtain Curcuma zedoaria powder. Composite extraction process: Adding water to the zedoary turmeric powder in a ratio of 1:1.5 and soaking the mixture in a sealed environment for 18 hours; extracting crude zedoary turmeric oil by steam distillation of the soaked zedoary turmeric powder; obtaining the crude zedoary turmeric oil; The crude zedoary turmeric oil is first pretreated by vacuum distillation, and then subjected to two-stage distillation using molecular distillation technology under conditions of a vacuum degree of less than 10 Pa and a temperature of less than 100° C., i.e., the first stage separates low-boiling point impurities, and the second stage collects the target fraction to obtain high-purity zedoary turmeric oil. The specific methods of the first stage separation and the second stage collection are as follows: First stage separation: Open the 150 short-range molecular distillation system, adjust the system temperature to 50°C and the pressure to 4-10KPa, add the crude zedoary oil to the feed port, adjust the flow rate to 2.5mL / min, and the scraper speed to 220r / min, and perform molecular distillation to collect the light zedoary oil; Second-stage collection: The molecular distillation system temperature was adjusted to 45°C, the pressure was adjusted to 50 Pa, the scraper speed was adjusted to 200 r / min, and the flow rate was 2.0 mL / min. The light-removed zedoary oil was subjected to primary distillation and the distillate was collected to obtain a zedoaryaryl oil product with a high content of geraniol and furanodiene.

[0024] Stable storage: Place high-purity zedoary oil into a brown glass container, fill it with nitrogen to replace the air, seal it, and store it in a refrigerator at 2-4°C away from light. Example 4

[0025] A method for increasing the content of geranone and furadiene in zedoary turmeric oil comprises the following steps: Stress treatment: At a zedoary turmeric cultivation base in Sichuan, a temperature-controlled greenhouse was constructed to treat low-temperature stress during the plant's peak growth period. Temperature control equipment within the greenhouse maintained the ambient temperature at 12°C for eight days. During the treatment period, the greenhouse temperature was monitored in real time to ensure it remained within the set range. Nutritional regulation: After the low temperature stress treatment, nutritional regulation was carried out. 22 kg of superphosphate and 16 kg of potassium sulfate were applied per mu, and the fertilization method was the same as in Examples 1 and 2. At the same time, the leaves of the zedoary turmeric were sprayed with 0.15% zinc sulfate solution and 0.06% borax solution every 15 days for 4 consecutive sprays. Pretreatment of Curcuma zedoaria: After the Curcuma zedoaria is mature and harvested, the rhizomes are washed and dried in a drying room to a moisture content of 7%. They are then crushed using a grinder and passed through a 40-mesh sieve to make Curcuma zedoaria powder. Composite extraction process: adding water to the zedoary turmeric powder in a ratio of 1:1.5 and soaking the mixture in a closed environment for 18 hours; extracting crude zedoary turmeric oil from the soaked zedoary turmeric powder by steam distillation to obtain crude zedoary turmeric oil; The crude zedoary turmeric oil is first pretreated by vacuum distillation, and then subjected to two-stage distillation using molecular distillation technology under conditions of a vacuum degree of less than 10 Pa and a temperature of less than 100° C., i.e., the first stage separates low-boiling point impurities, and the second stage collects the target fraction to obtain high-purity zedoary turmeric oil. The specific methods of the first stage separation and the second stage collection are as follows: First stage separation: Open the 150 short-range molecular distillation system, adjust the system temperature to 50°C and the pressure to 4-10KPa, add the crude zedoary oil to the feed port, adjust the flow rate to 2.5mL / min, and the scraper speed to 220r / min, and perform molecular distillation to collect the light zedoary oil; Second-stage collection: The molecular distillation system temperature was adjusted to 45°C, the pressure was adjusted to 50 Pa, the scraper speed was adjusted to 200 r / min, and the flow rate was 2.0 mL / min. The light-removed zedoary oil was subjected to primary distillation and the distillate was collected to obtain a zedoaryaryl oil product with a high content of geraniol and furanodiene.

[0026] Stable storage: Place high-purity zedoary oil into a brown glass container, fill it with nitrogen to expel all air, seal it, and store it in a refrigerator at 2-4°C away from light.

[0027] Specific experimental verification 1. Experimental Purpose The effectiveness of the method provided by the present invention for increasing the content of geraniol and furanodiene in zedoary turmeric oil was verified, the differences in the content of target components in zedoary turmeric oil after treatment in different embodiments were compared with those in the traditional method, and the extraction rate and purification effect of the method were evaluated.

[0028] 2. Experimental Materials and Instruments Experimental Materials: Control samples were collected from Curcuma rhizomes not treated with the present invention in Guangxi, Yunnan, and Sichuan. Curcuma rhizomes treated according to Examples 2-4 were also prepared. Chemical reagents such as petroleum ether, hydrophobic ionic liquids (e.g., 1-butyl-3-methylimidazolium hexafluorophosphate), zinc sulfate, borax, superphosphate, potassium sulfate, and sodium chloride were all of analytical grade. Carbon dioxide (food grade) was also used. Experimental instruments: 150 molecular distillation system, centrifuge, rotary evaporator, high performance liquid chromatography (HPLC), electronic balance, blast drying oven, constant temperature incubator, soil moisture meter, temperature and humidity recorder, etc. 3. Experimental Methods Determination of active ingredient content: Sample preparation: 1 g of the control sample and the Curcuma zedoaria powder treated in Examples 2-4 were taken, accurately weighed, placed in a stoppered conical flask, added with 20 mL of methanol, and ultrasonically extracted for 30 minutes. The mixture was filtered and the filtrate was diluted with methanol to an appropriate concentration as the test solution. Chromatographic conditions: High performance liquid chromatography was used with octadecylsilane bonded silica gel as the filler; the mobile phase was acetonitrile-water (gradient elution); the detection wavelength was 203 nm; the column temperature was 30°C; and the flow rate was 1.0 mL / min. Content determination: Accurately pipette 10 μL of the reference solution and the test solution respectively, inject them into the high performance liquid chromatograph, determine the peak area of ​​geraniol and furanodiene, and calculate the content according to the standard curve. Extraction rate calculation: accurately record the mass of the zedoary oil finally obtained in each embodiment and control experiment, and calculate the extraction rate according to the mass of the zedoary powder input according to the formula "extraction rate (%) = (mass of zedoary oil / mass of zedoary powder) × 100%". Comparative experiment: Traditional steam distillation control: Take the same mass of untreated Zedoaria rhizome as in the example, wash it and extract Zedoaria oil by steam distillation, record the extraction time and the mass of Zedoaria oil, and determine the contents of geraniol and furanodiene therein. Single extraction method control: Supercritical carbon dioxide extraction, microwave-assisted extraction, and ionic liquid-assisted extraction were selected to perform separate extraction experiments on untreated Curcuma zedoaria powder, setting the same parameters as in the composite extraction process of the present invention, recording the extraction results, and determining the content of the target components. IV. Experimental Procedure According to the above experimental material preparation requirements, complete the procurement and preparation of materials and reagents required for the experiment, and debug and calibrate the experimental instruments to ensure their normal operation. To conduct an active ingredient content determination experiment, test solutions of a control sample and samples of each example were first prepared, and then high performance liquid chromatography analysis was performed according to chromatographic conditions, and the peak area was recorded and the content was calculated. The extraction rates of zedoary turmeric oil in each embodiment and control experiment were calculated, and the effects of different treatment methods on the extraction rate were analyzed. Carry out comparative experiments, conduct experiments on traditional steam distillation method and single extraction method respectively, record experimental data and conduct measurement and analysis.

[0029] 5. Data Processing and Analysis 1. Data organization Categorize and organize the data on the zedoary oil extraction rate, geranone content, and furanodiene content for the control samples, traditional steam distillation samples, single extraction method samples, and samples from Examples 2-4, and enter them into an Excel spreadsheet. For example, create columns such as "Sample Type," "Extraction Rate (%)," "Geranone Content (%)," and "Furandiene Content (%)" to ensure data accuracy. 2. Calculate statistics Use Excel or SPSS software to calculate the mean and standard deviation for each data set. For the extraction rate, for example, for the control sample, calculate the mean of all replicate extraction rate data to reflect the central tendency of the data set; calculate the standard deviation to reflect the degree of dispersion. Similarly, perform the same calculations for geranone content and furanodienes content. 3. Analysis of variance (ANOVA) One-way analysis of variance (ANOVA) was performed using SPSS software to determine whether there were significant differences in the zedoary turmeric oil extraction yield, geranone content, and furandiene content among the different treatments (control sample, traditional method, single extraction method, and Examples 2-4). A significance level of 0.05 was set; if the P value was less than 0.05, the difference between the groups was considered significant; if the P value was greater than 0.05, the difference between the groups was considered insignificant. For example, when analyzing the difference in geranone content between Example 2 and the control sample, a P value of < 0.05 indicated that the treatment in Example 2 significantly increased the geranone content.

[0030] Expected Experimental Results 1. Comparison of active ingredient content Sample type Geraniol content (%) Furandiene content (%) Control samples 8.2 ± 0.5 5.1 ± 0.3 Traditional steam distillation sample 9.0 ± 0.6 5.5 ± 0.4 Supercritical carbon dioxide extraction single extraction sample 11.5 ± 0.8 7.2 ± 0.5 Microwave-assisted extraction of single sample 10.8 ± 0.7 6.8 ± 0.4 Ionic liquid-assisted extraction of single sample 12.0 ± 0.9 7.5 ± 0.6 Example 2 Sample 18.5 ± 1.2 12.3 ± 0.8 Example 3 Sample 17.8 ± 1.1 11.8 ± 0.7 Example 4 Sample 18.2 ± 1.0 12.0 ± 0.8 Comparison of the data reveals that the geraniol and furanodienes levels in the zedoary oils treated in Examples 2-4 were significantly higher than those in the control sample and the traditional steam distillation sample, and significantly higher than the target component levels in zedoary oils obtained using a single extraction method. This demonstrates that the stress treatment, nutritional regulation, and combined extraction process of the present invention synergistically promote the accumulation and extraction of active ingredients. 2. Extraction rate comparison Sample type Extraction rate (%) Control samples 3.2 ± 0.2 Traditional steam distillation sample 3.8 ± 0.3 Supercritical carbon dioxide extraction single extraction sample 4.5 ± 0.4 Microwave-assisted extraction of single sample 4.2 ± 0.3 Ionic liquid-assisted extraction of single sample 4.8 ± 0.5 Example 2 Sample 7.5 ± 0.6 Example 3 Sample 7.3 ± 0.5 Example 4 Sample 7.4 ± 0.6 The extraction rate of the composite extraction process of the present invention is significantly higher than that of the traditional steam distillation method and the single extraction method. The composite extraction process fully utilizes the advantages of each extraction method and extracts the zedoary turmeric oil more fully. 3. Purification effect High-performance liquid chromatography analysis and impurity detection showed that the impurity peak area of ​​the zedoary turmeric oil treated by the gradient purification process of the present invention was greatly reduced, and the purity reached more than 95%, which was significantly higher than the purity of the zedoary turmeric oil obtained by the traditional extraction method (about 80%), effectively improving the quality of the zedoary turmeric oil.

[0031] The present invention is provided as an example, not as a limitation of the embodiments. Those skilled in the art will appreciate that other variations or modifications may be made based on the above description. It is not necessary and impossible to enumerate all embodiments here, and obvious variations or modifications derived therefrom remain within the scope of protection of the present invention.

Claims

1. A method for increasing the content of geranone and furadiene in zedoary turmeric oil, characterized in that: The following steps are involved: Stress treatment: During the critical growth period of Curcuma zedoaria, moderate drought, low temperature or salt stress is used to activate the secondary metabolic pathway of Curcuma zedoaria and promote the accumulation of sesquiterpenes; Nutritional regulation: After stress treatment, increase the application of phosphorus, potassium fertilizers and trace elements such as zinc and boron, reduce nitrogen fertilizers, and optimize the ratio of active ingredients in the rhizomes; Curcuma zedoaria pretreatment: Wash and dry the rhizomes of Curcuma zedoaria to a moisture content of less than 10%, and grind them through a 40-60 mesh sieve to obtain Curcuma zedoaria powder, which provides a suitable raw material for subsequent extraction; Composite extraction process: Adding water to the zedoary turmeric powder in a ratio of 1:1.5 and soaking the mixture in a sealed environment for at least 18 hours; extracting crude zedoary turmeric oil by steam distillation of the soaked zedoary turmeric powder; The crude zedoary turmeric oil is first pretreated by vacuum distillation, and then subjected to two-stage distillation using molecular distillation technology under conditions of a vacuum degree of less than 10 Pa and a temperature of less than 100° C., i.e., the first stage separates low-boiling point impurities, and the second stage collects the target fraction to obtain high-purity zedoary turmeric oil; Stable storage: The purified zedoary oil is placed in a brown glass container, filled with nitrogen to replace the air, and then sealed. It is then stored in a refrigerator at 2-4°C away from light.

2. A method for increasing the content of geranone and furadiene in zedoary turmeric oil according to claim 1, characterized in that: The moderate drought stress in the adverse stress treatment is to control the soil moisture content at 40%-60% of the field holding capacity during the tuber swelling period, which lasts for 10-15 days.

3. A method for increasing the content of geranone and furadiene in zedoary turmeric oil according to claim 1, characterized in that: The low temperature stress of the adverse stress treatment is to control the environmental temperature at 10-15° C. for 7-10 days during the vigorous growth period.

4. A method for increasing the content of geranone and furadiene in zedoary turmeric oil according to claim 1, characterized in that: The salt stress in the adverse stress treatment is to apply a sodium chloride solution with a concentration of 0.2%-0.5% to the soil during the seedling stage, once every 10 days, for a total of 2-3 times.

5. A method for increasing the content of geranone and furadiene in zedoary turmeric oil according to claim 1, characterized in that: The nutritional regulation in the nutritional regulation is specifically as follows: applying 20-30 kg of superphosphate and 15-20 kg of potassium sulfate per mu, and spraying 0.1%-0.3% zinc sulfate solution and 0.05%-0.1% borax solution on the leaves once every 15 days, for a total of 3-4 times; the application amount of nitrogen fertilizer is controlled at 10-15 kg per mu.

6. A method for increasing the content of geranone and furadiene in zedoary turmeric oil according to claim 1, characterized in that: The specific method of the first stage separation in the composite extraction process is as follows: open the 150 short-range molecular distillation system, adjust the system temperature to 50°C and the pressure to 4-10Kpa, add the crude zedoary oil to the feed port, adjust the flow rate to 2.5mL / min, and the scraper speed to 220r / min, and perform molecular distillation to collect the light zedoary oil; The specific method for collecting the target fraction in the second stage is as follows: the distillation temperature of the molecular distillation system is adjusted to 45°C, the pressure is adjusted to 50Pa, the speed of the scraper is adjusted to 200r / min, and the flow rate is 2.0mL / min, the zedoary oil with light content is distilled in one stage, and the distillate is collected to obtain a zedoary oil product with high content of geraniol and furanodiene.