Method for extracting amomum tsao-ko essential oil

Abstract

The invention provides a method for extracting amomum tsao-ko essential oil. The method for extracting the amomum tsao-ko essential oil comprises the following steps: (1) crushing amomum tsao-ko fruits, and performing supercritical carbon dioxide extraction to obtain crude oil; (2) carrying out centrifugal treatment on the crude product oil to obtain amomum tsao-ko oil and oil-containing wax; and (3) mixing the oil-containing wax with ethanol, centrifuging, and concentrating the centrifuged supernatant liquid to obtain the amomum tsao-ko oil, and (4) mixing the amomum tsao-ko oil obtained in the step (2) and the step (3), and purifying through molecular distillation to obtain the amomum tsao-ko essential oil. The amomum tsao-ko essential oil extracted by the method disclosed by the invention is high in quality and has a remarkable improvement effect on constipation.

Description

Technical Field

[0001] The invention relates to the technical field of essential oil extraction, and in particular to a method for extracting tsaoko essential oil. Background Art

[0002] As the largest digestive and immune organ in the human body, the intestine is not only responsible for the absorption of nutrients and energy intake, but also regulates the immune system, which has an important impact on the health of the body from different angles and levels.

[0003] Tsaoko is a perennial herbaceous plant of the ginger family that is both medicinal and edible. The tsaoko essential oil extracted from tsaoko has anti-inflammatory, antioxidant, and digestion-promoting effects. The current tsaoko essential oil preparation process has significant defects, such as limited purity of the obtained product, residual solvents, poor color, and impure aroma, which leads to poor medicinal efficacy and is not conducive to the vigorous promotion and application of tsaoko essential oil. Therefore, it is necessary to develop a new tsaoko essential oil preparation process. Summary of the invention

[0004] In order to solve one of the above technical problems existing in the prior art, the present invention provides a method for extracting tsaoko essential oil. The tsaoko essential oil extracted by the method of the present invention can significantly improve its efficacy in intestinal regulation, especially in improving constipation.

[0005] The first aspect of the present invention provides a method for extracting tsaoko essential oil, comprising the following steps:

[0006] (1) crushing the tsaoko fruit and subjecting it to supercritical carbon dioxide extraction to obtain crude oil;

[0007] (2) centrifuging the crude oil to obtain tsaoko oil and oil-containing wax;

[0008] (3) mixing the oil-containing wax with ethanol and centrifuging the mixture, and concentrating the upper liquid after the centrifugation to obtain tsaoko oil;

[0009] (4) mixing the tsaoko oils obtained in step (2) and step (3), and purifying by molecular distillation to obtain tsaoko essential oil.

[0010] According to some embodiments of the present invention, in step (1), the tsaoko fruit is crushed into 20-60 meshes.

[0011] According to some embodiments of the present invention, in step (1), the pressure of the supercritical carbon dioxide extraction is 15 MPa to 20 MPa, for example, 15 MPa, 16 MPa, 17 MPa, 18 MPa, 19 MPa, 20 MPa or any value therebetween.

[0012] According to some embodiments of the present invention, in step (1), the temperature of the supercritical carbon dioxide extraction is 50-60°C, for example, 50°C, 52°C, 55°C, 58°C, 60°C or any value therebetween, preferably 55-60°C.

[0013] According to some embodiments of the present invention, in step (1), the supercritical carbon dioxide extraction time is 30 to 120 min, preferably 60 to 120 min, and more preferably 60 to 100 min.

[0014] According to some embodiments of the present invention, in step (2), the centrifugal rotation speed is 3500-4500 r / min, for example, 3500 r / min, 3800 r / min, 4000 r / min, 4200 r / min, 4500 r / min or any value therebetween; preferably, 3800-4200 r / min.

[0015] According to some embodiments of the present invention, in step (2), the centrifugation temperature is 0-10°C.

[0016] According to some embodiments of the present invention, in step (2), the centrifugation temperature is 3-8°C.

[0017] According to some embodiments of the present invention, in step (2), the centrifugation temperature is 4-6°C.

[0018] According to some embodiments of the present invention, in step (2), the centrifugation time is 10 to 20 minutes.

[0019] According to some embodiments of the present invention, in step (3), the amount of ethanol used is 5-10 times, for example, 5 times, 6 times, 7 times, 8 times, 9 times or 10 times the volume of the oil-containing wax.

[0020] According to some embodiments of the present invention, in step (3), the ethanol is selected from ethanol with a volume fraction of 95% or more.

[0021] According to some embodiments of the present invention, in step (3), the ethanol is ethanol with a volume fraction of 95%.

[0022] According to some embodiments of the present invention, in step (3), the centrifugal rotation speed is 3500-4500 r / min, for example, 3500 r / min, 3800 r / min, 4000 r / min, 4200 r / min, 4500 r / min or any value therebetween; preferably, 3800-4200 r / min.

[0023] According to some embodiments of the present invention, in step (3), the centrifugation temperature is 0-10°C.

[0024] According to some embodiments of the present invention, in step (3), the centrifugation temperature is 3-8°C.

[0025] According to some embodiments of the present invention, in step (3), the centrifugation temperature is 4-6°C.

[0026] According to some embodiments of the present invention, in step (3), the centrifugation time is 10 to 20 minutes.

[0027] According to some embodiments of the present invention, in step (3), the upper liquid after centrifugation includes ethanol and tsaoko oil.

[0028] According to some embodiments of the present invention, in step (3), the concentration temperature is 60-70°C, preferably 62-68°C.

[0029] According to some embodiments of the present invention, in step (4), the tsaoko oil is dried before molecular distillation. Preferably, the drying is performed using a molecular sieve. In the present invention, the molecular sieve includes but is not limited to Hβ molecular sieve, 4A molecular sieve, etc. In some embodiments, in step (4), 250 to 350 g of molecular sieve is added to every 1 kg of tsaoko oil. In some embodiments, in step (4), 280 to 320 g of molecular sieve is added to every 1 kg of tsaoko oil.

[0030] According to some embodiments of the present invention, in step (4), the feed temperature of the molecular distillation is 45-55°C, for example, 45°C, 48°C, 50°C, 52°C, 55°C or any value therebetween, preferably 50-55°C.

[0031] According to some embodiments of the present invention, in step (4), the feed flow rate of the molecular distillation is 450-550 mL / h, for example, 450 mL / h, 480 mL / h, 500 mL / h, 520 mL / h, 550 mL / h or any value therebetween.

[0032] According to some embodiments of the present invention, in step (4), the evaporation temperature of one section of the molecular distillation is 115-125°C, for example, 115°C, 118°C, 120°C, 122°C, 125°C or any value therebetween. In some embodiments, in step (4), the evaporation temperature of one section of the molecular distillation is 120-125°C.

[0033] According to some embodiments of the present invention, in step (4), the second stage evaporation temperature of the molecular distillation is 55-65°C, for example, 55°C, 58°C, 60°C, 62°C, 65°C or any value therebetween.

[0034] The second aspect of the present invention provides an essential oil of Amomum villosum, which is extracted by the method described in the first aspect of the present invention.

[0035] According to some embodiments of the present invention, the tsaoko essential oil is a brown-yellow clear liquid.

[0036] According to some embodiments of the present invention, the relative density (20°C / 20°C) of the tsaoko essential oil is 0.930-0.945.

[0037] According to some embodiments of the present invention, the refractive index (20°C±0.2°C) of the tsaoko essential oil is 1.475-1.488.

[0038] According to some embodiments of the present invention, the acid value (mg·KOH / mg) of the tsaoko essential oil is 16.5 to 17.5.

[0039] According to some embodiments of the present invention, the miscibility (20°C±0.2°C) of the tsaoko essential oil is as follows: 1 volume of tsaoko essential oil is dissolved in 20 volumes of 95% (volume fraction) ethanol solution.

[0040] According to some embodiments of the present invention, the miscibility (20°C±0.2°C) of the tsaoko essential oil is: 1 volume of tsaoko essential oil is insoluble in 20 volumes of water.

[0041] According to some embodiments of the present invention, the miscibility (20°C±0.2°C) of the tsaoko essential oil is: 1 volume of the tsaoko essential oil is insoluble in 50% to 95% (volume fraction, excluding 95%) ethanol solution.

[0042] The third aspect of the present invention provides a pharmaceutical composition, which includes the Amomum villosum essential oil described in the second aspect of the present invention.

[0043] According to some embodiments of the present invention, the pharmaceutical composition is an oral dosage form or a non-oral dosage form. The oral dosage form of the present invention includes but is not limited to solid oral dosage forms (such as enteric-coated tablets, dripping pills, oral tablets, chewable tablets, granules, powders or capsules) or liquid oral dosage forms (such as syrups or tinctures). Non-oral dosage forms of the present invention include but are not limited to administration by injection or other systemic routes, and the other systemic routes are such as transdermal administration or transmucosal administration (such as nasal, sublingual, oral, vaginal or rectal, by suppositories). Other routes of administration (such as those that can be used in veterinary applications) include enteral and parenteral delivery, including muscle, subcutaneous and / or intramedullary injection, and intrathecal injection, direct ventricular injection, intravenous injection, intraperitoneal injection, intranasal injection or intraocular injection.

[0044] According to some embodiments of the present invention, the pharmaceutical composition further comprises pharmaceutically acceptable excipients. The excipients of the present invention include, but are not limited to, calcium carbonate, calcium phosphate, calcium sulfate, sucrose, glucose, lactose, fructose, xylitol, sorbitol, starch, starch paste, cellulose derivatives, gelatin, polyvinyl pyrrolidone, sodium chloride, dextrin, stearic acid, magnesium stearate, calcium stearate, vegetable oil, polyethylene glycol, sterile phosphate buffered saline, saline, Ringer's solution, and combinations thereof.

[0045] The fourth aspect of the present invention provides use of the Amomum villosum essential oil described in the second aspect or the pharmaceutical composition described in the third aspect in the preparation of a medicament for preventing or treating constipation.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] 1. The method for extracting tsaoko essential oil of the present invention uses tsaoko fruits as raw materials, directly performs supercritical carbon dioxide extraction after crushing, and then subjecting the crude oil after extraction to processes such as centrifugation, molecular sieve treatment and concentration, so as to recover the tsaoko oil that is difficult to separate from the wax component in the crude oil, and high-quality tsaoko essential oil can still be obtained without complex process conditions such as acid extraction and enzymolysis.

[0048] 2. The present invention utilizes supercritical CO2 extraction combined with a molecular distillation purification process to effectively remove impurities such as macromolecular waxes, pigments, and colloids in the tsaoko essential oil; and before molecular distillation purification, a molecular sieve is used to dry the tsaoko oil, and the molecular sieve can also adsorb and remove residual small molecular impurities, thereby further improving the quality of the essential oil, improving the color and aroma of the essential oil, and increasing the purity.

[0049] 3. The tsaoko essential oil extracted by the method of the present invention can significantly improve constipation, has good effect and high safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 The present invention is a flowchart of a specific embodiment of the process for extracting tsaoko essential oil.

[0051] Figure 2 These are photos of the Amomum villosum essential oil obtained in Example 1 and Comparative Example 1 of the present application, wherein 1 represents Comparative Example 1 and 2 represents Example 1.

[0052] Figure 3 The results are compared in terms of the time to first black stool in mice after treatment with loperamide-induced constipation model using the tsaoko essential oils extracted from Example 1, Example 2, Comparative Example 1 and Comparative Example 2.

[0053] Figure 4 The blood toxicity results of the tsaoko essential oil extracted in Example 1. DETAILED DESCRIPTION

[0054] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments and drawings. The specific embodiments described herein are only used to explain the present invention and are not intended to constitute any limitation to the present invention.

[0055] The reagents used in the following experiments of the present invention, unless otherwise specified, are all conventional commercial products or reagents prepared by conventional methods, and the methods used in the experiments, unless otherwise specified, are all conventional experimental methods. The instruments used in the experiments, unless otherwise specified, can all be obtained through commercial channels.

[0056] The principle of using supercritical carbon dioxide to extract tsaoko essential oil is:

[0057] Under supercritical conditions, the supercritical fluid (carbon dioxide) is brought into contact with the substance to be separated (tsaoko fruit) so that it can selectively extract the components of polarity, boiling point and molecular weight in sequence. The dried tsaoko fruit contains oily substances. By controlling the extraction pressure and extraction temperature, the tsaoko oil, waxy substances and water in the tsaoko are extracted in sequence. Under high pressure, supercritical CO2 can penetrate into the tsaoko particles and flow the CO2 containing dissolved compounds to another device to reduce the pressure. Under lower pressure, CO2 becomes gaseous, and the compounds dissolved in it will reprecipitate. The separated tsaoko oil can remove CO2 from it through condensation and evaporation processes to obtain the tsaoko oil oil. The extracted tsaoko oil is a mixture of tsaoko oil, wax and water. Due to the large difference in density between the three, water can be placed in the bottom layer, wax in the middle layer, and tsaoko oil in the upper layer through high-speed centrifugation. Water and wax are removed to obtain pure tsaoko oil.

[0058] As a specific embodiment of the present invention, Figure 1 As shown, the method for extracting tsaoko essential oil comprises the following steps:

[0059] The tsaoko fruit is crushed and then subjected to supercritical carbon dioxide extraction to obtain crude oil;

[0060] The crude oil is centrifuged to obtain tsaoko oil 1, oil-containing wax and water;

[0061] The oil-containing wax is mixed with 95% ethanol by volume and then centrifuged to obtain an ethanol solution of wax and tsaoko oil; the ethanol solution containing tsaoko oil is concentrated to obtain tsaoko oil 2;

[0062] The obtained tsaoko oil 1 and tsaoko oil 2 are mixed with molecular sieves, and then molecular distillation is performed for purification to obtain tsaoko essential oil.

[0063] Example 1 Extraction of Amomum villosum essential oil

[0064] 1. Supercritical CO2 extraction:

[0065] The Amomum villosum fruit is crushed into appropriate particle size (20-60 mesh) to increase its surface area for easy extraction.

[0066] The crushed tsaoko particles are placed in a supercritical CO2 extractor and pressurized to a supercritical state to obtain primary oil. Extraction parameters: 18MPa / 55℃; separation parameters: 6.5MPa / 60℃; time: 90min.

[0067] 2. Centrifugation:

[0068] The primary oil obtained by supercritical CO2 extraction was centrifuged at 4000 r / min, 5°C, and 15 min. After centrifugation, the upper layer of grass fruit oil was separated and recorded as grass fruit oil 1, and the lower layer was waxy.

[0069] 3. Oil-containing wax dissolution:

[0070] The waxy layer obtained in step 2 above, which is difficult to separate from the tsaoko oil, is added with 5-10 times the volume of 95% (volume fraction) ethanol, fully dissolved and then centrifuged at 4000r / min, 5°C, 15min. After centrifugation, the tsaoko oil containing ethanol in the upper layer is separated and set aside.

[0071] 4. Concentration:

[0072] The ethanol-containing amomum villosum oil obtained in step 3 is concentrated at a concentration temperature of 65° C. The obtained concentrated solution is recorded as amomum villosum oil 2.

[0073] 5. Combined with grass fruit oil:

[0074] Combine the tsaoko oil 1 obtained in step 2 and the tsaoko oil 2 obtained in step 4.

[0075] 6. Remove moisture:

[0076] Molecular sieves were used to remove excess water from the combined tsaoko oil in step 5, with 300 g of molecular sieves added for every 1 kg of tsaoko oil.

[0077] 7. Molecular distillation:

[0078] The tsaoko oil after removing water in step 6 was subjected to molecular distillation treatment, with a feed temperature of 50°C, a flow rate of 500 mL / h, a first evaporation temperature of 120°C, a second evaporation temperature of 60°C, and the second distillation portion was collected to obtain tsaoko essential oil (appearance and clarity as shown in FIG. Figure 2 shown).

[0079] Embodiment 2 Extraction of Amomum villosum essential oil

[0080] 1. Supercritical CO2 extraction:

[0081] The Amomum villosum fruit is crushed into appropriate particle size (20-60 mesh) to increase its surface area for easy extraction.

[0082] The crushed tsaoko particles are placed in a supercritical CO2 extractor and pressurized to a supercritical state to obtain primary oil. Extraction parameters: 18MPa / 55℃; separation parameters: 6.5MPa / 60℃; time: 90min.

[0083] 2. Centrifugation:

[0084] The primary oil obtained by supercritical CO2 extraction was centrifuged at 4000 r / min, 5°C, and 15 min. After centrifugation, the upper layer of grass fruit oil was separated and recorded as grass fruit oil 1, and the lower layer was waxy.

[0085] 3. Oil-containing wax dissolution:

[0086] The waxy layer obtained in step 2 above, which is difficult to separate from the tsaoko oil, is added with 5-10 times the volume of 95% (volume fraction) ethanol, fully dissolved and then centrifuged at 4000r / min, 5°C, 15min. After centrifugation, the tsaoko oil containing ethanol in the upper layer is separated and set aside.

[0087] 4. Concentration:

[0088] The ethanol-containing amomum villosum oil obtained in step 3 is concentrated at a concentration temperature of 65° C. The obtained concentrated solution is recorded as amomum villosum oil 2.

[0089] 5. Combined with grass fruit oil:

[0090] Combine the tsaoko oil 1 obtained in step 2 and the tsaoko oil 2 obtained in step 4.

[0091] 6. Remove moisture:

[0092] Molecular sieves were used to remove excess water from the combined tsaoko oil in step 5, with 300 g of molecular sieves added for every 1 kg of tsaoko oil.

[0093] 7. Molecular distillation:

[0094] The tsaoko oil after water removal in step 6 is subjected to molecular distillation treatment, with a feed temperature of 45° C., a flow rate of 500 mL / h, a first-stage evaporation temperature of 115° C., a second-stage evaporation temperature of 60° C., and the second-stage distillation portion is collected to obtain tsaoko essential oil.

[0095] Example 3 Extraction of Amomum villosum essential oil

[0096] 1. Supercritical CO2 extraction:

[0097] The Amomum villosum fruit is crushed into appropriate particle size (20-60 mesh) to increase its surface area for easy extraction.

[0098] The crushed tsaoko particles are placed in a supercritical CO2 extractor and pressurized to a supercritical state to obtain primary oil. Extraction parameters: 18MPa / 55℃; separation parameters: 6.5MPa / 60℃; time: 90min.

[0099] 2. Centrifugation:

[0100] The primary oil obtained by supercritical CO2 extraction was centrifuged at 4000 r / min, 5°C, and 15 min. After centrifugation, the upper layer of grass fruit oil was separated and recorded as grass fruit oil 1, and the lower layer was waxy.

[0101] 3. Oil-containing wax dissolution:

[0102] The waxy layer obtained in step 2 above, which is difficult to separate from the tsaoko oil, is added with 5-10 times the volume of 95% (volume fraction) ethanol, fully dissolved and then centrifuged at 4000r / min, 5°C, 15min. After centrifugation, the tsaoko oil containing ethanol in the upper layer is separated and set aside.

[0103] 4. Concentration:

[0104] The ethanol-containing amomum villosum oil obtained in step 3 is concentrated at a concentration temperature of 65° C. The obtained concentrated solution is recorded as amomum villosum oil 2.

[0105] 5. Combined with grass fruit oil:

[0106] Combine the tsaoko oil 1 obtained in step 2 and the tsaoko oil 2 obtained in step 4.

[0107] 6. Molecular distillation:

[0108] The combined tsaoko oil in step 5 is subjected to molecular distillation treatment, with a feed temperature of 50° C., a flow rate of 500 mL / h, a first-stage evaporation temperature of 120° C., a second-stage evaporation temperature of 60° C., and the second-stage distillation portion is collected to obtain tsaoko essential oil.

[0109] Comparative Example 1 Extraction of Amomum villosum essential oil

[0110] Supercritical CO2 extraction:

[0111] The Amomum villosum fruit is crushed into appropriate particle size (20-60 mesh) to increase its surface area for easy extraction.

[0112] The crushed tsaoko particles were placed in a supercritical CO2 extractor and pressurized to a supercritical state. The extraction parameters were 18MPa / 55°C, separation parameters were 6.5MPa / 60°C, and the time was 90min. The tsaoko essential oil (appearance and clarity were as shown in Table 1) was obtained. Figure 2 shown).

[0113] Comparative Example 2 Extraction of Amomum villosum essential oil

[0114] 1. Supercritical CO2 extraction:

[0115] The Amomum villosum fruit is crushed into appropriate particle size (20-60 mesh) to increase its surface area for easy extraction.

[0116] The crushed tsaoko particles are placed in a supercritical CO2 extractor and pressurized to a supercritical state to obtain primary oil. Extraction parameters: 18MPa / 55℃; separation parameters: 6.5MPa / 60℃; time: 90min.

[0117] 2. Remove moisture:

[0118] The excess water in the primary oil obtained in step 1 is removed by using molecular sieves, and 300 g of molecular sieves are added for every 1 kg of primary oil.

[0119] 3. Molecular distillation:

[0120] The initial oil after removing water in step 2 is subjected to molecular distillation treatment, with a feed temperature of 50° C., a flow rate of 500 mL / h, a first-stage evaporation temperature of 120° C., a second-stage evaporation temperature of 60° C., and the second-stage distillation portion is collected to obtain tsaoko essential oil.

[0121] Experiment 1: Physical and chemical index test

[0122] According to the standard of "T / CACM 1543.1-2023 Amomum villosum essential oil", the appearance, clarity and relative density of the Amomum villosum essential oil prepared in the above examples and comparative examples were measured. 20 20 , refractive index Nd 20 The physical and chemical indicators such as acid value, solubility in ethanol and water, aroma and flavor quality, Pb content, As content, etc. were tested, and the results are shown in Table 1.

[0123] Table 1

[0124]

[0125] As can be seen from Table 1 above, the methods of Examples 1, 2 and 3 of the present invention are first extracted by supercritical CO2, and then the crude oil obtained by extraction is separated and recovered by a centrifugal process to recover the insoluble wax components, and then molecular distillation is performed after molecular sieve treatment (or molecular distillation is directly performed without molecular sieve treatment). Compared with the Tsaoko essential oil obtained by supercritical CO2 extraction in Comparative Example 1 and the method of supercritical CO2 extraction in Comparative Example 2 without centrifugal separation and recovery of wax, the Tsaoko essential oil obtained in Examples 1, 2 and 3 is lighter in color, better in clarity, greater in solubility in 95% ethanol solution, and better in aroma and flavor quality.

[0126] Experiment 2 Effect of Amomum villosum essential oil on symptom relief of loperamide-induced constipation model in mice

[0127] Fifty 6-8 week old male C57BL / 6J mice were divided into 5 groups, with 10 mice in each group.

[0128] The first group was the model group, which was gavaged with normal saline in the first week and loperamide hydrochloride solution at a dose of 50 mg / kg in the second week.

[0129] The second group was treatment group 1, which was gavaged with the essential oil of Amomum villosum extracted in Example 1 at a dose of 200 μL / kg every day for two consecutive weeks; in the second week, the mice were gavaged with loperamide hydrochloride solution at a dose of 50 mg / kg.

[0130] The third group was treatment group 2, which was gavaged with the essential oil of Amomum villosum extracted in Example 2 at a dose of 200 μL / kg every day for two consecutive weeks; in the second week, the mice were gavaged with loperamide hydrochloride solution at a dose of 50 mg / kg.

[0131] The fourth group was treatment group 3, which was gavaged with the essential oil of Amomum villosum extracted from comparative example 1 at a dose of 200 μL / kg every day for two consecutive weeks; in the second week, the mice were gavaged with loperamide hydrochloride solution at a dose of 50 mg / kg.

[0132] The fifth group is treatment group 4, which was gavaged with the essential oil of Amomum villosum extracted from comparative example 2 at a dose of 200 μL / kg every day for two consecutive weeks; in the second week, the mice were gavaged with loperamide hydrochloride solution at a dose of 50 mg / kg.

[0133] After modeling and treatment, mice in each group were gavaged with 0.25 mL of activated carbon solution. The time was counted from the gavage of activated carbon solution and the time of the first discharge of black stool was recorded. Preparation of activated carbon solution: 5 g of gum arabic was added to 100 mL of distilled water, and 5 g of activated carbon powder was added after boiling. After boiling for 3 times, it was placed at room temperature to cool, and distilled water was added to the total volume of the solution to 100 mL. Finally, the activated carbon solution was obtained and stored in a refrigerator at 4 °C for later use.

[0134] The time of the first black stool of mice in each group is shown in Figure 3 Compared with other treatment groups, the time for the mice to pass the first black stool was shorter after treatment with the Tsaoko essential oil extracted in Examples 1 and 2. In particular, the Tsaoko essential oil extracted in Example 1 can significantly shorten the time for the model mice to pass the first black stool, indicating that it has a significant effect on improving constipation.

[0135] Experiment 3 Evaluation of the blood toxicity of the essential oil of Amomum villosum extracted in Example 1 in mice

[0136] Fifteen male C57BL / 6J mice aged 6-8 weeks were divided into three groups, with 5 mice in each group. The first group was a normal group, which was gavaged with normal saline for two consecutive weeks. The second group was a low-dose group, which was gavaged with the essential oil of Amomum villosum extracted in Example 1 at a dose of 200 μL / kg every day for two consecutive weeks. The third group was a high-dose group, which was gavaged with the essential oil of Amomum villosum extracted in Example 1 at a dose of 400 μL / kg every day for two consecutive weeks. After the experiment, the eyeballs were removed and blood was collected in anticoagulant tubes for routine blood tests. The results of the blood toxicity investigation are shown in Figure 4 , indicating that the tsaoko essential oil extracted in Example 1 of the present invention has no blood toxicity.

[0137] The technical solution of the present invention is not limited to the above-mentioned specific embodiments. All technical variations made according to the technical solution of the present invention fall within the protection scope of the present invention.

Claims

1. A method for extracting tsaoko essential oil, comprising the following steps: (1) crushing the tsaoko fruit and subjecting it to supercritical carbon dioxide extraction to obtain crude oil; (2) centrifuging the crude oil to obtain tsaoko oil and oil-containing wax; (3) mixing the oil-containing wax with ethanol and centrifuging the mixture, and concentrating the upper liquid after the centrifugation to obtain tsaoko oil; (4) mixing the tsaoko oils obtained in step (2) and step (3), and purifying by molecular distillation to obtain tsaoko essential oil.

2. The method according to claim 1, characterized in that: In step (1), the conditions for supercritical carbon dioxide extraction are: pressure 15MPa-20MPa, and / or temperature 50-60°C, preferably 55-60°C; and / or time 30-120min, preferably 60-100min.

3. The method according to claim 1 or 2, characterized in that: In step (2), the centrifugal speed is 3500-4500 r / min, preferably 3800-4200 r / min; and / or the centrifugal temperature is 0-10°C, preferably 3-8°C; and / or the centrifugal time is 10-20 min.

4. The method according to any one of claims 1 to 3, characterized in that: In step (3), the amount of ethanol used is 5-10 times the volume of the oil-containing wax; and / or the ethanol is ethanol with a volume fraction of more than 95%.

5. The method according to any one of claims 1 to 4, characterized in that: In step (3), the centrifugal rotation speed is 3500-4500 r / min, preferably 3800-4200 r / min; and / or the centrifugal temperature is 0-10°C, preferably 3-8°C; and / or the centrifugal time is 10-20 min; and / or the upper layer liquid after centrifugation includes ethanol and tsaoko oil; and / or the concentration temperature is 60-70°C, preferably 62-68°C.

6. The method according to any one of claims 1 to 5, characterized in that: In step (4), drying is performed before molecular distillation purification, preferably using molecular sieves for drying; preferably, the molecular sieves include Hβ molecular sieves and / or 4A molecular sieves; Preferably, 250-350 g, preferably 280-320 g, of molecular sieve is added to every 1 kg of grass fruit oil.

7. The method according to any one of claims 1 to 6, characterized in that: In step (4), the feed temperature of the molecular distillation is 45 to 55° C., preferably 50 to 55° C.; and / or the feed flow rate is 450 to 550 mL / h; and / or the first evaporation temperature is 115 to 125° C., preferably 120 to 125° C.; and / or the second evaporation temperature is 55 to 65° C.

8. An essential oil of Amomum villosum, which is extracted by the method according to any one of claims 1 to 7; Preferably, the tsaoko essential oil is a brown-yellow clear liquid; Preferably, the relative density (20°C / 20°C) of the tsaoko essential oil is 0.930 to 0.945; Preferably, the refractive index (20°C ± 0.2°C) of the tsaoko essential oil is 1.475 to 1.488; Preferably, the acid value (mg·KOH / mg) of the tsaoko essential oil is 16.5 to 17.5; Preferably, the miscibility (20°C±0.2°C) of the tsaoko essential oil is: 1 volume of tsaoko essential oil is dissolved in 20 volumes of 95% (volume fraction) ethanol solution.

9. A pharmaceutical composition comprising the tsaoko essential oil according to claim 8 and pharmaceutically optional auxiliary materials.

10. Use of the tsaoko essential oil according to claim 8 or the pharmaceutical composition according to claim 9 in the preparation of a medicament for preventing or treating constipation.

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