Preparation method of ganoderma lucidum spore oil with liver regulating effect

Abstract

The invention relates to the technical field of preparation methods of nutritional products, in particular to a preparation method of ganoderma lucidum spore oil with a liver regulating effect. Comprising the steps of preparing ganoderma lucidum spore powder, extracting ganoderma lucidum spore oil and enriching the ganoderma lucidum spore oil. Ganoderma lucidum blocks are treated through an ultrasonic extraction technology, ganoderma lucidum spore powder is prepared through a spray drying method, n-hexyl alcohol is used for soaking extraction, high-purity ganoderma lucidum spore oil is obtained through vacuum concentration and molecular distillation, and residual solvent is removed through film evaporation. Active ingredients such as triterpenoids and unsaturated fatty acid are further enriched by using supercritical CO2 extraction; the extraction efficiency and the purity of active ingredients of the ganoderma lucidum spore oil are remarkably improved, the obtained ganoderma lucidum spore oil has remarkable effects in the aspects of reducing liver toxicity indexes (ALT and AST) and enhancing antioxidant enzyme activity (SOD and GSH-Px), the operation controllability is high, the ganoderma lucidum spore oil is suitable for large-scale industrial application, and technical support is provided for research and development of liver protection drugs and functional food.

Description

Technical Field

[0001] The invention relates to the technical field of a method for preparing a nutritional product, and in particular to a method for preparing ganoderma lucidum spore oil having a liver regulating effect. Background Art

[0002] Ganoderma lucidum spore oil is a precious natural oil extracted from Ganoderma lucidum spores, rich in triterpenoids, unsaturated fatty acids, steroids and a variety of biologically active ingredients. As one of the core active ingredients of Ganoderma lucidum, Ganoderma lucidum spore oil has significant biological effects in regulating immune function, anti-oxidation, anti-inflammatory, anti-tumor and promoting liver repair. In the field of traditional Chinese medicine and functional food, Ganoderma lucidum spore oil has attracted widespread attention for its excellent liver regulating effect.

[0003] As the metabolic center of the human body, the liver plays an important role in detoxification, protein synthesis and bile production. However, due to changes in modern lifestyles and increased environmental pollution, the incidence of liver diseases has been increasing year by year. Liver-related diseases such as fatty liver, cirrhosis and drug-induced liver injury seriously threaten human health, and there is an urgent need to develop functional products with efficient liver protection.

[0004] At present, the preparation methods of Ganoderma lucidum spore oil mainly include cold pressing, organic solvent extraction, supercritical CO2 extraction and enzymatic hydrolysis. However, these methods have certain limitations in extraction efficiency, ingredient retention rate and environmental friendliness. For example, although the traditional cold pressing method can retain some active ingredients, the extraction efficiency is low; the organic solvent extraction method may introduce harmful residues; the supercritical CO2 extraction method is environmentally friendly and efficient, but the cost is relatively high and the equipment is complex. In addition, the thick wall structure of the outer wall of Ganoderma lucidum spores forms a natural barrier to the release of ingredients, which further increases the technical difficulty of preparing Ganoderma lucidum spore oil.

[0005] In order to overcome the above problems, a method for preparing Ganoderma lucidum spore oil was developed which can efficiently release the contents of Ganoderma lucidum spores while taking into account extraction efficiency and ingredient retention rate. This can not only better tap the potential of Ganoderma lucidum spore oil in regulating liver function, but also promote the widespread application of Ganoderma lucidum-related products to meet market demand. Summary of the invention

[0006] In view of the above problems, the present invention aims to provide a method for preparing Ganoderma lucidum spore oil having liver regulating effect, comprising the following steps:

[0007] S1 Preparation of Ganoderma lucidum spore powder;

[0008] Ganoderma lucidum is crushed to obtain Ganoderma lucidum blocks, the Ganoderma lucidum blocks are subjected to ultrasonic treatment, the Ganoderma lucidum liquid concentration and the ultrasonic power are used as parameters to obtain the optimal Ganoderma lucidum liquid concentration and the ultrasonic power, the Ganoderma lucidum blocks are extracted using the optimal Ganoderma lucidum liquid concentration and the optimal ultrasonic power, after extraction, centrifugation is performed to obtain Ganoderma lucidum extract, and the Ganoderma lucidum extract is spray-dried to obtain Ganoderma lucidum spore powder;

[0009] Preparation of S2 Ganoderma lucidum spore oil:

[0010] The ganoderma lucidum spore powder is soaked in fatty alcohol, and the fatty alcohol soaking liquid is extracted after soaking, and concentrated to obtain a concentrated liquid, and then the concentrated liquid is subjected to solvent fractionation to obtain ganoderma lucidum spore oil;

[0011] Enrichment of S3 Ganoderma lucidum spore oil:

[0012] The ganoderma spore oil is subjected to thin film evaporation to remove the fluoride solvent to obtain refined ganoderma spore oil, and then the refined ganoderma spore oil is subjected to supercritical CO2 extraction to obtain the final product of ganoderma spore oil.

[0013] Furthermore, the fatty alcohol is n-hexanol.

[0014] Furthermore, in step S1, the concentration of the Ganoderma lucidum liquid is in the range of 1-10 g / ml, and the ultrasonic power is in the range of 80-400 W.

[0015] Furthermore, in the step S1, for the concentration of the Ganoderma lucidum liquid, a target concentration of 3 g / ml is adopted, and for the ultrasonic power, a target power of 150 W is adopted.

[0016] Furthermore, in step S1, obtaining the optimal Ganoderma lucidum liquid concentration and the optimal ultrasonic power includes:

[0017] S11 Determination of the concentration of Ganoderma lucidum liquid;

[0018] Taking the concentration of Ganoderma lucidum liquid as a variable, five different concentration points of 2g / ml, 4g / ml, 6g / ml, 8g / ml and 10g / ml were selected to perform ultrasonic extraction on Ganoderma lucidum block material, and the leaching rate of the extract was used as an evaluation index to draw a concentration-leaching rate curve, and the concentration value corresponding to the highest point of the concentration-leaching rate curve was taken as the optimal Ganoderma lucidum liquid concentration;

[0019] S12 Determination of ultrasonic power;

[0020] Taking ultrasonic power as a variable, taking ultrasonic power of 100W, 150W, 200W, 250W, 300W, 350W and 400W, a total of seven power points, ultrasonic extraction of Ganoderma lucidum block was carried out, taking the leaching rate of the extract as an evaluation index, drawing a power-leaching rate curve, and taking the power value corresponding to the highest point of the power-leaching rate curve as the optimal ultrasonic power;

[0021] S13 Synergistic optimization of Ganoderma lucidum liquid concentration and ultrasonic power;

[0022] Taking the optimal Ganoderma lucidum concentration and the optimal ultrasonic power as the initial values, the response surface methodology was used to construct a mathematical model with the leaching rate as the response variable and the Ganoderma lucidum concentration and the ultrasonic power as the independent variables. The mathematical model was solved to obtain the synergistic optimal solution of the optimal Ganoderma lucidum concentration and the optimal ultrasonic power.

[0023] Furthermore, in the step S2, the mass ratio of the Ganoderma lucidum spore powder to the fatty alcohol is 1:6.

[0024] Furthermore, in step S2, the soaking treatment temperature is 50-90° C., and the soaking time is 8-12 hours.

[0025] Furthermore, the concentration is reduced pressure concentration, and the fractionation is molecular distillation.

[0026] Further, the supercritical CO2 extraction comprises:

[0027] The refined Ganoderma lucidum spore oil is placed in a supercritical CO2 extraction device, and the refined Ganoderma lucidum spore oil is extracted under supercritical CO2 conditions to obtain a supercritical CO2 extract, and the supercritical CO2 extract is post-processed to obtain a final product, which is the Ganoderma lucidum spore oil with liver regulating effect.

[0028] Beneficial Effects

[0029] The present invention provides a method for preparing ganoderma lucidum spore oil having liver regulating effect, and the efficient preparation of ganoderma lucidum spore oil is achieved by optimizing the extraction process and enrichment means, and the beneficial effects are as follows:

[0030] 1. Improve extraction efficiency: The present invention adopts ultrasonic extraction combined with spray drying technology. During the preparation process of Ganoderma lucidum spore powder, the leaching rate and active ingredient content of Ganoderma lucidum spore powder are significantly improved through the synergistic optimization of the optimal Ganoderma lucidum liquid concentration (3g / ml) and the optimal ultrasonic power (150W), thereby improving the raw material utilization rate.

[0031] 2. Enhance the purity and activity of Ganoderma lucidum spore oil: In the preparation of Ganoderma lucidum spore oil, non-target components are effectively removed through immersion treatment with fatty alcohols (such as n-hexanol) and molecular distillation fractionation technology. Ganoderma lucidum spore oil is further refined through thin film evaporation and supercritical CO2 extraction, which greatly improves the purity of key active ingredients such as triterpenoids and unsaturated fatty acids in the product.

[0032] 3. Optimize product biological activity: Experimental data show that the Ganoderma lucidum spore oil prepared by the method of the present invention exhibits significant liver protection in a hepatotoxic induced mouse model. Compared with the comparative example, the serum ALT and AST levels of mice in the Ganoderma lucidum spore oil group of the embodiment were significantly reduced, while the activities of liver antioxidant enzymes (SOD, GSH-Px) were significantly increased, and the content of lipid peroxides (MDA) was significantly reduced, indicating that the product has significant advantages in regulating liver function and enhancing antioxidant capacity.

[0033] 4. Green and environmentally friendly process: The present invention adopts green processes such as ultrasonic extraction, fatty alcohol immersion and supercritical CO2 extraction, which greatly reduces the use of organic solvents, reduces environmental pollution, and meets the requirements of modern green production.

[0034] 5. Strong process controllability: By optimizing the concentration of Ganoderma lucidum liquid and ultrasonic power parameters and adopting a multi-stage separation and refining process, the method provided by the present invention has strong process controllability and is suitable for large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 It is a schematic diagram of the method flow of the present invention.

[0036] Figure 2 These are photos of mouse liver and HE photos of liver tissue in the mouse liver poison comparison experiment of the present invention.

[0037] Figure 3 It is a schematic diagram of the results of serum ALT and AST levels in the mouse liver poison comparison experiment of the present invention.

[0038] Figure 4 It is a schematic diagram of the results of antioxidant enzyme activity and MDA content in liver tissue of the mouse liver poison comparison experiment of the present invention. DETAILED DESCRIPTION

[0039] In order to deepen the understanding of the present invention, the present invention will be further described in detail below in conjunction with examples. The examples are only used to explain the present invention and do not constitute a limitation on the protection scope of the present invention.

[0040] Example

[0041] according to Figure 1 As shown, this embodiment provides step S1 for preparing Ganoderma lucidum spore powder, which specifically includes:

[0042] Step S1, preparation of Ganoderma lucidum spore powder:

[0043] Take 500 grams of fresh fruiting bodies of cultivated Ganoderma lucidum, wash them clean and remove impurities.

[0044] The cleaned Ganoderma lucidum fruiting bodies were dried in an oven at 60° C. for 12 hours to obtain dried Ganoderma lucidum blocks.

[0045] The dried Ganoderma lucidum blocks were placed in a crusher for crushing, and 200 grams of Ganoderma lucidum powder with a particle size of less than 40 meshes was obtained by screening.

[0046] Ultrasonic treatment of Ganoderma lucidum blocks:

[0047] To prepare Ganoderma lucidum liquid, take 10 grams of dried Ganoderma lucidum blocks and add 30 milliliters of distilled water (the concentration of Ganoderma lucidum liquid is 3g / ml).

[0048] The ultrasonic extractor containing the Ganoderma lucidum block and distilled water was placed on the ultrasonic generator, and the ultrasonic power was set to 150 W and the processing time was 30 minutes.

[0049] After the ultrasonic treatment, the mixture was allowed to stand at room temperature for 10 minutes.

[0050] Extraction and spray drying:

[0051] The mixed solution after ultrasonic treatment was centrifuged at a speed of 3000 rpm for 20 minutes to separate and obtain a clear Ganoderma lucidum extract.

[0052] The Ganoderma lucidum extract was spray dried by a spray drying device, and the spray drying conditions were set as a feed temperature of 150° C. and a discharge temperature of 80° C. to obtain 50 g of Ganoderma lucidum spore powder.

[0053] Step S2: Preparation of Ganoderma lucidum spore oil

[0054] Fatty alcohol soaking treatment:

[0055] Take 50 grams of Ganoderma lucidum spore powder and add 300 ml of n-hexanol in a mass ratio of 1:6.

[0056] The mixture was placed in a constant temperature water bath, the temperature was controlled at 70°C, and the soaking time was 10 hours.

[0057] Concentration of extract:

[0058] After the soaking is completed, the mixed solution is vacuum filtered at a filtration temperature of 40° C. to remove solid impurities. The filtered n-hexanol soaking solution is concentrated in a vacuum concentration device to a volume of about 50 ml to obtain a concentrated solution.

[0059] Solvent fractionation:

[0060] The concentrated solution was placed in a molecular distillation device, and the molecular distillation conditions were set at a temperature of 120 °C and a vacuum degree of 1 × 10 - 3 After the fractionation, the low boiling point components were collected to obtain 30 ml of Ganoderma lucidum spore oil.

[0061] Step S3: Enrichment of Ganoderma lucidum spore oil

[0062] Thin film evaporation:

[0063] 30 ml of the obtained Ganoderma lucidum spore oil was placed in a thin film evaporation device, and the evaporation temperature was set to 50° C. and the evaporation pressure was set to 500 Pa.

[0064] After the evaporation is completed, the fluoride solvent is removed to obtain 28 ml of refined Ganoderma lucidum spore oil.

[0065] Supercritical CO2 extraction: Place 28 ml of refined Ganoderma lucidum spore oil in a supercritical CO2 extraction device.

[0066] The supercritical CO2 extraction conditions were set to a pressure of 300 bar, a temperature of 40°C, a flow rate of 10 g / min, and an extraction time of 2 hours.

[0067] After the extraction is completed, the supercritical CO2 extract is collected and subjected to a reduced pressure solvent removal treatment to obtain the final product - 25 ml of Ganoderma lucidum spore oil with liver regulating effects.

[0068] Product Analysis:

[0069] Through high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) analysis, it was confirmed that the final product contained significant levels of triterpenoids, polysaccharides and other bioactive ingredients, and had good anti-liver damage activity.

[0070] Comparative Example A

[0071] Step S1: Preparation of Ganoderma Lucidum Spore Powder

[0072] Preparation and crushing of ganoderma lucidum block: 500 g of wild ganoderma lucidum fruiting bodies were taken, cleaned and impurities were removed. The cleaned ganoderma lucidum fruiting bodies were naturally dried at room temperature for 24 hours to obtain dried ganoderma lucidum blocks. The dried ganoderma lucidum blocks were crushed using a common crusher and sieved to obtain 200 g of ganoderma lucidum powder with a particle size of less than 60 mesh.

[0073] Soaking and extraction of ganoderma lucidum block: add 10 g of dried ganoderma lucidum block to 50 ml of ethanol (70%), soak for 2 hours, and place at room temperature. After the extraction, filter with filter paper to obtain ganoderma lucidum extract. Naturally dry the ganoderma lucidum extract under normal pressure to obtain 50 g of ganoderma lucidum spore powder.

[0074] Step S2: Preparation of Ganoderma Lucidum Spore Oil

[0075] Ethanol soaking treatment: Take 50 grams of Ganoderma lucidum spore powder, add 200 ml of ethanol at a mass ratio of 1:4. Place the mixture on a shaker and shake and soak at room temperature for 6 hours.

[0076] Concentration of the extract: After the soaking is completed, the mixed solution is filtered through a filter to remove solid impurities. The filtered ethanol soaking solution is concentrated to 25 ml using a rotary evaporator at normal pressure to obtain a concentrated solution.

[0077] Solvent fractionation: The concentrated solution was placed in a common distillation apparatus and distilled under normal pressure to separate and obtain 20 ml of Ganoderma lucidum spore oil.

[0078] Step S3: Enrichment of Ganoderma lucidum spore oil

[0079] Simple filtration: Filter the obtained 20 ml of Ganoderma lucidum spore oil through activated carbon to remove impurities and obtain 18 ml of refined Ganoderma lucidum spore oil.

[0080] Normal pressure extraction: 18 ml of refined Ganoderma lucidum spore oil was placed in a normal pressure extraction device and extracted with petroleum ether for 1 hour. After the extraction, the solvent was removed under reduced pressure to obtain the final product - 15 ml of Ganoderma lucidum spore oil.

[0081] Product analysis: High performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) were used to confirm the content of active ingredients in the final product.

[0082] Comparative Example B

[0083] Step S1: Preparation of Ganoderma Lucidum Spore Powder

[0084] Preparation and crushing of Ganoderma lucidum block: Take 400 g of cultivated Ganoderma lucidum fruiting bodies, clean them and remove impurities. Dry the cleaned Ganoderma lucidum fruiting bodies in a microwave dryer at 120°C for 6 hours to obtain dried Ganoderma lucidum blocks. Grind the dried Ganoderma lucidum blocks using a ball mill and screen to obtain 180 g of Ganoderma lucidum powder with a particle size of less than 80 mesh.

[0085] Hot reflux extraction of Ganoderma lucidum block: 10 g of dried Ganoderma lucidum block was added to 40 ml of methanol and subjected to hot reflux extraction for 3 hours, with the reflux temperature controlled at 80°C. After the extraction, the Ganoderma lucidum extract was filtered with filter paper to obtain the Ganoderma lucidum extract. The Ganoderma lucidum extract was dried in a microwave-assisted dryer to obtain 45 g of Ganoderma lucidum spore powder.

[0086] Step S2: Preparation of Ganoderma Lucidum Spore Oil

[0087] Methanol soaking treatment: Take 45 grams of Ganoderma lucidum spore powder, add 225 ml of methanol at a mass ratio of 1:5, place the mixture in a reflux extractor, and perform hot reflux extraction for 2 hours.

[0088] Concentration of the extract: After the extraction is completed, the mixed solution is filtered through a Buchner funnel to remove solid impurities. The filtered methanol soaking solution is concentrated to 30 ml in a rotary evaporator to obtain a concentrated solution.

[0089] Solvent fractionation: The concentrated solution was placed in a common distillation apparatus and distilled under normal pressure to separate and obtain 25 ml of Ganoderma lucidum spore oil.

[0090] Step S3: Enrichment of Ganoderma lucidum spore oil

[0091] Refrigerated centrifugation: Place 25 ml of Ganoderma lucidum spore oil in a refrigerated centrifuge, centrifuge at 4°C, 4000 rpm for 30 minutes, and separate 22 ml of refined Ganoderma lucidum spore oil.

[0092] Ethanol extraction: 22 ml of refined Ganoderma lucidum spore oil was mixed with 200 ml of ethanol and subjected to ethanol extraction for 1.5 hours. After the extraction, the solvent was removed under reduced pressure to obtain the final product, 18 ml of Ganoderma lucidum spore oil.

[0093] Product analysis: High performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) analysis confirmed that the content of active ingredients in the final product was relatively high.

[0094] Comparative Example C

[0095] Step S1: Preparation of Ganoderma Lucidum Spore Powder

[0096] Preparation and pulverization of Ganoderma lucidum block: 600 g of cultivated Ganoderma lucidum fruiting bodies were taken, cleaned and impurities removed. The cleaned Ganoderma lucidum fruiting bodies were dried in an oven at 80°C for 8 hours to obtain dried Ganoderma lucidum blocks. The dried Ganoderma lucidum blocks were ground using a grinder and sieved to obtain 220 g of Ganoderma lucidum powder with a particle size of less than 50 mesh.

[0097] Cold soaking extraction of ganoderma lucidum block: 10 g of dried ganoderma lucidum block was added to 60 ml of water, placed in an ice bath, and cold soaked for 12 hours. After the extraction, vacuum filtration was performed to obtain a ganoderma lucidum extract. The ganoderma lucidum extract was naturally dried under normal pressure to obtain 55 g of ganoderma lucidum spore powder.

[0098] Step S2: Preparation of Ganoderma Lucidum Spore Oil

[0099] Water-alcohol mixed soaking treatment: Take 55 grams of Ganoderma lucidum spore powder, add 275 milliliters of water-alcohol mixed solvent (50% ethanol) according to the mass ratio of 1:5, place the mixture in a constant temperature shaker, shake and soak at room temperature for 8 hours.

[0100] Concentration of the extract: After the soaking is completed, the mixed solution is filtered through gauze to remove solid impurities. The filtered water-alcohol soaking solution is concentrated to 35 ml in a rotary evaporator to obtain a concentrated solution.

[0101] Solvent fractionation: The concentrated solution was placed in a molecular distillation device, and the molecular distillation conditions were set at a temperature of 100°C and a vacuum degree of 5×10-3 Pa. After the fractionation was completed, the middle boiling point component was collected to obtain 28 ml of Ganoderma lucidum spore oil.

[0102] Step S3: Enrichment of Ganoderma lucidum spore oil

[0103] High temperature distillation: 28 ml of Ganoderma lucidum spore oil was placed in a high temperature distillation device, and the distillation temperature was set to 70°C and the distillation pressure was set to 600 Pa. After the distillation was completed, 24 ml of refined Ganoderma lucidum spore oil was obtained.

[0104] Ethyl acetate extraction: 24 ml of refined Ganoderma lucidum spore oil was mixed with 150 ml of ethyl acetate and extracted with ethyl acetate for 2 hours. After the extraction, the solvent was removed under reduced pressure to obtain the final product - 20 ml of Ganoderma lucidum spore oil.

[0105] Product analysis: High performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) were used to confirm the content of active ingredients in the final product.

[0106] Mouse liver toxicant comparison experiment

[0107] In order to verify the effect of the Ganoderma lucidum spore oil prepared in Example 1 of the present invention on regulating liver function and enhancing liver metabolism, the following toxicity experiment was designed. The experiment included one experimental group (Example 1) and three comparative experimental groups (Comparative Examples A, B, and C).

[0108] The effects of different preparation methods on the effect of Ganoderma lucidum spore oil were evaluated by comparing the liver function metabolism capacity of each group of mice under toxicant induction.

[0109] 1. Experimental Materials and Methods

[0110] 1. Experimental Animals

[0111] Sixty healthy adult male mice were selected and randomly divided into four groups, with 15 mice in each group.

[0112] Animal source: Laboratory-bred mice, weighing 18-22 grams.

[0113] Housing conditions: temperature 22±2℃, relative humidity 55±10%, 12 h light / 12 h dark cycle, free access to food and water.

[0114] 2. Grouping and processing

[0115] Experimental group: Ganoderma lucidum spore oil prepared in Example 1 (hereinafter referred to as "Example oil").

[0116] Control group A: Ganoderma lucidum spore oil prepared by the method of comparative example A (hereinafter referred to as "comparative example A oil").

[0117] Control group B: Ganoderma lucidum spore oil prepared by the method of comparative example B (hereinafter referred to as "comparative example B oil").

[0118] Control group C: Ganoderma lucidum spore oil prepared in comparative example C (hereinafter referred to as "comparative example C oil").

[0119] Blank group: No Ganoderma lucidum spore oil was taken, only poison treatment was performed.

[0120] 3. Oil administration

[0121] Administration: Oral gavage.

[0122] Dosage: Each mouse was orally administered 200 mg / kg body weight of Ganoderma lucidum spore oil daily.

[0123] Dosage cycle: Take for two weeks continuously.

[0124] 4. Toxic Induction

[0125] Poison selection: Carbon tetrachloride (CCl4).

[0126] Induction dose: Each mouse was intraperitoneally injected with CCl4 oil solution (CCl4: vegetable oil = 1:10, dose of 10 mL / kg body weight).

[0127] Induction time: CCl4 induction was performed starting from the 15th day of taking the oil, twice a week, for a total of 4 weeks.

[0128] 5. Sampling and testing

[0129] Sampling time: 24 hours after the last CCl4 induction.

[0130] Detection indicators:

[0131] Serum alanine aminotransferase (ALT) level.

[0132] Serum aspartate aminotransferase (AST) level.

[0133] Antioxidant enzyme activities (SOD, GSH-Px) in liver tissue.

[0134] Lipid peroxide content (MDA) in liver tissue.

[0135] Liver histopathological analysis (HE staining), such as Figure 2 shown.

[0136] 6. Statistics

[0137] The data are expressed as mean ± SD.

[0138] Comparisons among groups were performed using one-way analysis of variance (ANOVA) followed by the Tukey-Kramer method.

[0139] The statistical significance level was set at P < 0.05.

[0140] 2. Experimental Results

[0141] Table 1 Serum ALT and AST levels

[0142] Group ALT(U / L)±SD AST(U / L)±SD Blank Group 150.2±12.5 180.3±15.1 Comparative Example A Oil Group 140.5±10.8 170.6±14.2 Comparative Example B Oil Group 135.7±11.2 165.4±13.7 Comparative Example C Oil Group 130.3±9.8 160.1±12.9 Example Oil Group 90.4±8.5* 110.2±10.3*

[0143] P<0.05, Example oil group vs. blank group, * indicates statistically significant difference compared with the control group. Figure 3 shown.

[0144] Table 2 Antioxidant enzyme activities and MDA content in liver tissue

[0145]

[0146] P<0.05, Example oil group vs. blank group, * indicates statistically significant difference compared with the control group. Figure 4 shown.

[0147] From Table 1 to Table 2, Figure 3 to Figure 4 It can be seen that the Ganoderma lucidum spore oil prepared by the methods of comparative examples A, B, and C can reduce serum ALT and AST levels and increase antioxidant enzyme activity, which proves that Ganoderma lucidum spore oil has the effect of protecting the liver. However, after mice took the Ganoderma lucidum spore oil prepared by the method of the present invention, the serum ALT and AST levels were reduced to a higher degree than those of the three comparative examples, and the antioxidant enzyme activity was higher, which can prove that the preparation method of the present invention can more effectively preserve the effective components of Ganoderma lucidum spore oil compared with the methods of the three comparative examples.

[0148] Further, from Figure 2 It can be seen that the liver tissue of mice in the blank group showed obvious fibrosis, the comparative example A group also showed fibrosis (the degree was lower than that of the blank group), and the comparative example B group also showed early fibrosis (the degree was lower than that of the blank group and the comparative example A group), which proved that Ganoderma lucidum spore oil has the effect of protecting the liver; it can be further seen that the liver tissue of mice in the embodiment group (the method of the present invention group) did not show obvious fibrosis, which proved that the Ganoderma lucidum spore oil prepared by the method of the present invention retains more effective ingredients.

[0149] The above shows and describes the basic principles, main features and advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A method for preparing Ganoderma lucidum spore oil having liver regulating effect, characterized in that: The following steps are involved: S1. Preparation of Ganoderma lucidum spore powder; Ganoderma lucidum is crushed to obtain Ganoderma lucidum blocks, the Ganoderma lucidum blocks are subjected to ultrasonic treatment, the Ganoderma lucidum liquid concentration and the ultrasonic power are used as parameters to obtain the optimal Ganoderma lucidum liquid concentration and the ultrasonic power, the Ganoderma lucidum blocks are extracted using the optimal Ganoderma lucidum liquid concentration and the optimal ultrasonic power, after extraction, centrifugation is performed to obtain Ganoderma lucidum extract, and the Ganoderma lucidum extract is spray-dried to obtain Ganoderma lucidum spore powder; S2. Preparation of Ganoderma lucidum spore oil: The ganoderma lucidum spore powder is soaked in fatty alcohol, and the fatty alcohol soaking liquid is extracted after soaking, and concentrated to obtain a concentrated liquid, and then the concentrated liquid is subjected to solvent fractionation to obtain ganoderma lucidum spore oil; S3. Enrichment of Ganoderma lucidum spore oil: The ganoderma spore oil is subjected to thin film evaporation to remove the fluoride solvent to obtain refined ganoderma spore oil, and then the refined ganoderma spore oil is subjected to supercritical CO2 extraction to obtain the final product of ganoderma spore oil.

2. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 1, characterized in that: The fatty alcohol is n-hexanol.

3. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 2, characterized in that: In the step S1, the concentration of the ganoderma lucidum liquid is in the range of 1-10 g / ml, and the ultrasonic power is in the range of 80-400 W.

4. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 3, characterized in that: In the step S1, the target concentration of the ganoderma lucidum liquid is 3 g / ml, and the target power of the ultrasonic wave is 150 W.

5. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 4, characterized in that: In the step S1, obtaining the optimal ganoderma lucidum liquid concentration and the optimal ultrasonic power comprises: S11 Determination of the concentration of Ganoderma lucidum liquid; Taking the concentration of Ganoderma lucidum liquid as a variable, five different concentration points of 2g / ml, 4g / ml, 6g / ml, 8g / ml and 10g / ml were selected to perform ultrasonic extraction on Ganoderma lucidum block material, and the leaching rate of the extract was used as an evaluation index to draw a concentration-leaching rate curve, and the concentration value corresponding to the highest point of the concentration-leaching rate curve was taken as the optimal Ganoderma lucidum liquid concentration; S12 Determination of ultrasonic power; Taking ultrasonic power as a variable, taking ultrasonic power of 100W, 150W, 200W, 250W, 300W, 350W and 400W, a total of seven power points, ultrasonic extraction of Ganoderma lucidum block was carried out, taking the leaching rate of the extract as an evaluation index, drawing a power-leaching rate curve, and taking the power value corresponding to the highest point of the power-leaching rate curve as the optimal ultrasonic power; S13 Synergistic optimization of Ganoderma lucidum liquid concentration and ultrasonic power; Taking the optimal Ganoderma lucidum concentration and the optimal ultrasonic power as the initial values, the response surface methodology was used to construct a mathematical model with the leaching rate as the response variable and the Ganoderma lucidum concentration and the ultrasonic power as the independent variables. The mathematical model was solved to obtain the synergistic optimal solution of the optimal Ganoderma lucidum concentration and the optimal ultrasonic power.

6. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 5, characterized in that: In the step S2, the mass ratio of the ganoderma lucidum spore powder to the fatty alcohol is 1:

6.

7. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 6, characterized in that: In the step S2, the soaking treatment temperature is 50-90°C and the soaking time is 8-12h.

8. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 7, characterized in that: The concentration is reduced pressure concentration, and the fractionation is molecular distillation.

9. The method for preparing Ganoderma lucidum spore oil having liver regulating effect according to claim 8, characterized in that: The supercritical CO2 extraction comprises: The refined Ganoderma lucidum spore oil is placed in a supercritical CO2 extraction device, and the refined Ganoderma lucidum spore oil is extracted under supercritical CO2 conditions to obtain a supercritical CO2 extract, and the supercritical CO2 extract is post-processed to obtain a final product, which is the Ganoderma lucidum spore oil with liver regulating effect.

Images