Extraction method of ganoderma lucidum spore oil and microcapsule prepared from ganoderma lucidum spore oil

Abstract

The invention discloses an extraction method of ganoderma lucidum spore oil. The extraction method comprises the following steps: (1) pretreatment; (2) carrying out superfine grinding to prepare partially wall-broken ganoderma lucidum spore powder; (3) enzymolysis treatment: preparing a ganoderma lucidum spore powder suspension; (4) carrying out centrifugal separation, and respectively collecting an oil phase, a water phase and a slag phase, the oil phase being ganoderma lucidum spore oil; the microcapsule containing the ganoderma lucidum spore oil is prepared by taking a water phase obtained in the extraction process and egg yolk phospholipoprotein as wall materials. The method can solve the technical problems of insufficient yield and easy oxidative degradation in the production and processing of the ganoderma lucidum spore oil.

Description

Technical Field

[0001] The invention relates to the technical field of plant oil extraction, in particular to a method for extracting ganoderma lucidum spore oil and microcapsules prepared therefrom. Background Art

[0002] Ganoderma lucidum spore oil has multiple pharmacological effects such as anti-oxidation, improving immunity, and inhibiting the physiological activity of tumor cells. It can also induce or promote the phagocytosis of macrophages, enhance the activity of natural killer (NK) cells, promote the production of interleukin 2 (IL-2) and interferon by helper cells (Th), increase the transformation rate of lymphocytes, and promote the formation of immunoglobulins, thereby comprehensively improving the body's non-specific immune function. Ganoderma lucidum spore oil can also regulate human immunity in both directions, regulate the proportion of T cell subsets in tumor patients, increase the activity of NK cells and the activities of phagocytes, IL-2, tumor necrosis factor, etc., and increase the transformation rate of lymphocytes. At the same time, it also has the effects of inhibiting the physiological activity of tumor cells and preventing the regeneration and metastasis of cancer cells.

[0003] There are many methods for extracting Ganoderma lucidum spore oil. The traditional hot pressing method is due to the poor heat resistance of Ganoderma lucidum spore oil, which can easily lead to the loss of nutrients and heat-sensitive components. It also faces the challenge of low product yield. The organic solvent extraction method is complex and may pollute the environment. Although the supercritical carbon dioxide extraction method is effective, it is costly and difficult to promote to large-scale production. The yield and quality of Ganoderma lucidum spore oil are related to the degree of spore wall breaking, the extraction method, and the oxidative degradation during the crushing, extraction and storage processes. Therefore, how to extract Ganoderma lucidum spore oil efficiently and environmentally friendly and achieve its stable storage is a key technical problem that needs to be solved in the field of spore oil processing technology. Summary of the invention

[0004] In view of the above problems existing in the prior art, the present invention provides a method for extracting ganoderma spore oil and microcapsules prepared therefrom, which can solve the technical problems of insufficient yield and easy oxidation and degradation in the production and processing of ganoderma spore oil.

[0005] The technical solution of the present invention is as follows:

[0006] The first object of the present invention is to provide a method for extracting Ganoderma lucidum spore oil, the extraction method comprising the following steps:

[0007] (1) Pretreatment: Ganoderma lucidum spore powder is sieved through a 30-40 mesh vibrating screen to remove impurities, and vacuum-dried at low temperature until the moisture content is less than 8 wt %;

[0008] (2) Ultrafine grinding: The impurity-removed Ganoderma lucidum spore powder is ground by ultra-low temperature vibration mill to obtain partially broken Ganoderma lucidum spore powder;

[0009] (3) Enzymatic hydrolysis: resuspending the partially broken spore powder in pure water, then adding neutral protease to perform enzymatic hydrolysis to obtain a Ganoderma lucidum spore powder suspension;

[0010] (4) Centrifugal separation: The Ganoderma lucidum spore powder suspension is subjected to centrifugal separation to collect the oil phase, water phase and residue phase respectively. The oil phase is the Ganoderma lucidum spore oil.

[0011] In one embodiment of the present invention, in step (1), the vacuum low-temperature drying treatment conditions are: temperature 50-60°C, time 2-4h, vacuum degree -0.06 to -0.08MPa.

[0012] In one embodiment of the present invention, in step (2), the pulverizing temperature is -38 to -28°C, and the pulverizing time is 30 min to 1 h; nitrogen protection is provided during the pulverizing process.

[0013] In one embodiment of the present invention, in step (3), the mass ratio of spore powder to pure water during the resuspension process is 1:5-10.

[0014] In one embodiment of the present invention, in step (3), the amount of neutral protease used relative to the Ganoderma lucidum spore powder is 2000-5000 U / g; and the enzymatic hydrolysis conditions are: enzymatic hydrolysis treatment at 45-50° C. for 2-4 hours.

[0015] In one embodiment of the present invention, in step (4), centrifugation is performed using an automatic slag discharge disc centrifuge; the conditions for centrifugal separation are: rotation speed 3000-9500 rpm, flow rate 1-3 t / h, back pressure 0.2-0.3 MPa.

[0016] The second object of the present invention is to provide a Ganoderma lucidum spore oil obtained by the above extraction method.

[0017] The third object of the present invention is to provide a microcapsule containing the above-mentioned Ganoderma lucidum spore oil, and the preparation method of the microcapsule is:

[0018] (1) mixing the aqueous phase obtained in the process of extracting the ganoderma spore oil with the egg yolk phospholipid protein in a vacuum emulsification tank, adjusting the pH to 7-8, stirring and mixing, and then pumping the oil phase obtained in the process of extracting the ganoderma spore oil into the vacuum emulsification tank for emulsification;

[0019] (2) The emulsified material is spray-dried to obtain microcapsule powder.

[0020] In one embodiment of the present invention, in step (1), the mass ratio of the aqueous phase to the egg yolk phospholipid protein is 10:1-2; the emulsification is stirred for 15-30 minutes under the conditions of a vacuum degree of -0.06-0.08 MPa, a stirring speed of 23-38 rpm, and an emulsification speed of 2800-3600 rpm.

[0021] In one embodiment of the present invention, in step (2), the spray drying conditions are: vacuum degree -50 to -2000 Pa, inlet temperature 110 to 120°C, and outlet temperature 55 to 60°C.

[0022] The beneficial technical effects of the present invention are:

[0023] The invention adopts ultrafine pulverization to effectively break the wall of ganoderma lucidum spore powder, and on this basis, utilizes protease enzymatic hydrolysis to hydrolyze the protein on the spore shell, further destroys the integrity of the spore shell wall, improves the oil extraction rate and releases bioactive peptides with antioxidant activity. Therefore, ultrafine pulverization, enzymatic hydrolysis and centrifugal deoiling technology are combined to improve the extraction rate of ganoderma lucidum spore oil and reduce the oxidation probability of the spore oil.

[0024] The egg yolk phospholipid protein adopted by the present invention is prepared from egg yolk powder through granulation, supercritical CO2 extraction, separation and other processes. The main components are protein (≥40.0g / 100g) and phospholipid (≥24.0g / 100g). The phospholipid protein is a protein-phospholipid complex obtained by removing the lipid droplet core from the lipoprotein in the egg yolk. The phospholipid protein has good emulsification and is used for the preparation of spore oil microcapsules. The phospholipid protein can simulate the characteristics of the yolk ball in the natural egg yolk to protect the spore oil and improve its oxidative stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a flow chart for the extraction of Ganoderma lucidum spore oil and its microcapsule preparation. DETAILED DESCRIPTION

[0026] The present invention is described in detail below in conjunction with the accompanying drawings and embodiments.

[0027] Embodiment 1:

[0028] A method for extracting ganoderma lucidum spore oil comprises the following steps:

[0029] (1) Pretreatment: Select high-quality Ganoderma lucidum spore powder, sieve the powder through a 40-mesh vibrating screen to remove impurities, and vacuum dry it at a temperature of 60° C. and a vacuum degree of -0.06 MPa for 4 hours until the moisture content is less than 8%;

[0030] (2) Ultrafine grinding: The impurity-removed Ganoderma lucidum spore powder is ground using a nitrogen-filled ultra-low temperature vibration mill at a grinding temperature of -28°C for 30 minutes to obtain partially broken spore powder;

[0031] (3) Enzymatic hydrolysis: partially spore powder with broken wall was resuspended in pure water at a mass ratio of 1:5, neutral protease was added at an addition amount of 2000 U / g (relative to partially spore powder with broken wall), and enzymatic hydrolysis was performed at 45° C. for 2 h to obtain a Ganoderma lucidum spore powder suspension;

[0032] (4) Centrifugal separation: The spore powder suspension is subjected to automatic slag-discharging disc centrifugal separation to collect the oil phase, water phase and slag phase respectively. The centrifugal conditions are: speed 3000 rpm, flow rate 1 t / h, back pressure 0.2 MPa; the oil phase is Ganoderma lucidum spore oil;

[0033] A method for preparing microcapsules containing the above-mentioned Ganoderma lucidum spore oil is as follows:

[0034] The aqueous phase separated and collected in the above step (4) is pumped into a vacuum emulsification tank, and egg yolk phospholipid protein is added at a mass ratio of 10:1, the pH is adjusted to 7, and stirred and mixed (stirring speed is 23 rpm), and then the oil phase separated and collected in the above step (4) is pumped into a vacuum emulsification tank, and emulsified for 30 minutes under the conditions of vacuum degree -0.06 MPa and emulsification speed of 2800 rpm; the emulsified material is spray-dried under the conditions of vacuum degree -50 Pa, inlet temperature 110° C., and outlet temperature 55° C. to obtain microcapsule powder.

[0035] Embodiment 2:

[0036] A method for extracting ganoderma lucidum spore oil comprises the following steps:

[0037] (1) Pretreatment: Select high-quality Ganoderma lucidum spore powder, sieve the powder through a 40-mesh vibrating screen to remove impurities, and vacuum dry it at a temperature of 60° C. and a vacuum degree of -0.06 MPa for 4 hours until the moisture content is less than 8%;

[0038] (2) Ultrafine grinding: The impurity-removed Ganoderma lucidum spore powder is ground using a nitrogen-filled ultra-low temperature vibration mill at a grinding temperature of -28°C for 30 minutes to obtain partially broken spore powder;

[0039] (3) Enzymatic hydrolysis: partially spore powder with broken wall was resuspended in pure water at a mass ratio of 1:10, neutral protease was added at an addition amount of 5000 U / g (relative to partially spore powder with broken wall), and enzymatic hydrolysis was performed at 45° C. for 2 h to obtain a Ganoderma lucidum spore powder suspension;

[0040] (4) Centrifugal separation: The spore powder suspension is subjected to automatic slag-discharging disc centrifugal separation to collect the oil phase, water phase and slag phase respectively. The centrifugal conditions are: speed 9500 rpm, flow rate 3 t / h, back pressure 0.3 MPa; the oil phase is Ganoderma lucidum spore oil;

[0041] A method for preparing microcapsules containing the above-mentioned Ganoderma lucidum spore oil is as follows:

[0042] The aqueous phase separated and collected in the above step (4) is pumped into a vacuum emulsification tank, and egg yolk phospholipid protein is added at a mass ratio of 10:2, the pH is adjusted to 7, and the mixture is stirred and mixed (stirring speed is 38 rpm). Then, the oil phase separated and collected in the above step (4) is pumped into a vacuum emulsification tank, and emulsified for 15 minutes under the conditions of vacuum degree -0.06 MPa and emulsification speed of 3600 rpm; the emulsified material is spray-dried under the conditions of vacuum degree -2000 Pa, inlet temperature 110° C., and outlet temperature 55° C. to obtain microcapsule powder.

[0043] Example 3

[0044] A method for extracting ganoderma lucidum spore oil comprises the following steps:

[0045] (1) Pretreatment: Select high-quality Ganoderma lucidum spore powder, sieve the powder through a 30-mesh vibrating screen to remove impurities, and vacuum dry it at a temperature of 50° C. and a vacuum degree of -0.08 MPa for 3 hours until the moisture content is less than 8%;

[0046] (2) Ultrafine grinding: The impurity-free Ganoderma lucidum spore powder is ground using a nitrogen-filled ultra-low temperature vibration mill at a grinding temperature of -38°C for 30 minutes to obtain partially broken spore powder;

[0047] (3) Enzymatic hydrolysis: partially spore powder with broken wall was resuspended in pure water at a mass ratio of 1:8, neutral protease was added at an addition amount of 4000 U / g (relative to partially spore powder with broken wall), and enzymatic hydrolysis was performed at 50° C. for 2 h to obtain a Ganoderma lucidum spore powder suspension;

[0048] (4) Centrifugal separation: The spore powder suspension is separated by an automatic slag-discharging disc centrifuge to collect the oil phase, water phase and slag phase respectively. The centrifugal conditions are: speed 5000 rpm, flow rate 2 t / h, back pressure 0.3 MPa; the oil phase is Ganoderma lucidum spore oil;

[0049] A method for preparing microcapsules containing the above-mentioned Ganoderma lucidum spore oil is as follows:

[0050] The aqueous phase separated and collected in the above step (4) is pumped into a vacuum emulsification tank, and egg yolk phospholipid protein is added at a mass ratio of 10:2, and the pH is adjusted to 7. After stirring and mixing (stirring speed is 30 rpm), the oil phase separated and collected in the above step (4) is pumped into a vacuum emulsification tank, and emulsified for 20 minutes under the conditions of vacuum degree -0.08 MPa and emulsification speed of 3000 rpm; the emulsified material is spray-dried under the conditions of vacuum degree -150 Pa, inlet temperature 120° C., and outlet temperature 60° C. to obtain microcapsule powder.

[0051] Comparative Example 1

[0052] Refer to Example 1, the only difference is that the ultrafine grinding in step (2) is omitted, and the other steps and parameters remain unchanged.

[0053] Comparative Example 2

[0054] Refer to Example 1, the only difference is that the enzymatic hydrolysis treatment in step (3) is omitted, and the other steps and parameters remain unchanged.

[0055] Comparative Example 3

[0056] Refer to Example 1, the only difference is that the preparation of microcapsules is omitted, and other steps and parameters remain unchanged.

[0057] Comparative Example 4

[0058] Refer to Example 1, the only difference is that the aqueous phase is replaced by an equal amount of water during the preparation of the microcapsules.

[0059] Test example:

[0060] The extraction rate of the Ganoderma lucidum spore oil prepared in Examples 1-3 and Comparative Examples 1-4 was calculated, the triterpene content of the spore oil and the polypeptide content and antioxidant activity in the aqueous phase were detected, and the peroxide value of the microcapsules after storage at 60° C. for 12 days was detected. The results are shown in Table 1 below.

[0061] (1) The spore oil extraction rate is calculated according to the following formula:

[0062]

[0063] (2) The triterpene content in spore oil was determined according to the method in Appendix A of the group standard T / JZLZ 005-2019 Jinzhai Ganoderma Lucidum Spore Oil.

[0064] (3) The peptide content in the aqueous phase separated by centrifugal deoiling was determined according to the method in Appendix B of GB / T 22492-2008 soybean peptide powder.

[0065] (4) The antioxidant activity of peptides in the aqueous phase separated by centrifugal deoiling was determined by referring to the ABTS method in the DPPH and ABTS method in GB / T 39100-2020 for the antioxidant activity of peptides. + Free radical scavenging method was used for determination.

[0066] (5) Evaluation of storage stability of spore oil microcapsules:

[0067] The prepared spore oil microcapsules were placed in a 60°C oven for 12 days and then the peroxide value was determined. The method is as follows: First, 0.5 g of microcapsule powder was accurately weighed and dissolved in 5 mL of deionized water. Next, 0.3 mL of the sample solution was taken and 1.5 mL of a 3:1 (v / v) mixed solvent of isooctane and isopropanol was added to it. After mixing evenly, the mixture was centrifuged at 2500 g for 2 min. Take 0.2mL of supernatant and mix with 2.8mL of methanol-butanol mixture (2:1, v / v), add 15μL of 0.3g / mL ammonium thiocyanate solution and 15μL of ferrous ion solution (0.6mL of 0.144mol / L ferrous sulfate solution and 0.132mol / L barium chloride + 0.4M hydrochloric acid mixed solution are taken into a 1.5ml plastic centrifuge tube, vortex mixed for 30s, and centrifuged at 5000rpm for 5min), let stand for 20min after mixing, and measure the absorbance at a wavelength of 510nm with a spectrophotometer. According to the standard curve of cumene hydroperoxide (y=1.8482x-0.0593, R 2 =0.9946), and the absorbance was converted into peroxide value.

[0068] Table 1 Analysis of the extraction rate of Ganoderma lucidum spore oil and the peroxide value in microcapsules after high temperature accelerated storage

[0069]

[0070]

[0071] Embodiments 1 to 3 of the present invention combine ultrafine grinding, enzymatic treatment and centrifugal deoiling to achieve synergistic effect as a whole, increase the spore oil extraction rate and the triterpene content in the spore oil, and further utilize the antioxidant effect of the polypeptide in the aqueous phase and the emulsification and embedding effect of the egg yolk phospholipid protein to microencapsulate and embed the spore oil for protection, thereby improving the storage stability of the spore oil powder.

[0072] By comparing the results in Example 1 with Comparative Example 1 and Table 1 thereof, it can be seen that if only enzymatic treatment is performed without ultrafine grinding treatment, the extraction rate will be greatly reduced, but the triterpene content of the obtained spore oil remains at a high level, but the lipid peroxide value of the microcapsule powder does not change significantly. This result shows that enzymatic treatment has a certain oil extraction effect, but is far inferior to the treatment method combining ultrafine grinding, enzymatic treatment and centrifugal deoiling.

[0073] By comparing the results in Example 1 and Comparative Example 2 and Table 1 thereof, it can be seen that the spore oil extraction rate will be reduced to a certain extent when only ultrafine grinding treatment is used without enzymatic hydrolysis treatment, and the peroxide value of the microcapsule powder in Comparative Example 2 after storage will be higher than that in Example 1. This result shows that enzymatic hydrolysis treatment can not only effectively improve the spore oil extraction rate, but also release some antioxidant peptides to improve the oxidative stability of lipids in the microencapsulated powder.

[0074] By comparing Example 1 with Comparative Example 3 and the results in Table 1, it can be seen that the spore oil after extraction is not microencapsulated, and the storage stability is poor, which is not conducive to its long-term storage.

[0075] By comparing the results in Example 1 and Comparative Example 4 and Table 1 thereof, it can be seen that after the aqueous phase is replaced with an equal amount of water during the preparation of the microcapsules, the peroxide value of the microcapsule powder is greatly increased, indicating that the antioxidant polypeptides released by the enzymatic hydrolysis during the extraction of Ganoderma lucidum spore oil can significantly inhibit the increase in the peroxide value of the spore oil microcapsule powder.

[0076] The embodiments provided above are not intended to limit the scope of the present invention, and the steps described are not intended to limit the execution order thereof. Those skilled in the art may make obvious improvements to the present invention in combination with existing common knowledge, which also fall within the scope of protection defined by the claims of the present invention.

Claims

1. A method for extracting Ganoderma lucidum spore oil, characterized in that: The extraction method comprises the following steps: (1) Pretreatment: Ganoderma lucidum spore powder is sieved through a 30-40 mesh vibrating screen to remove impurities, and vacuum-dried at low temperature until the moisture content is less than 8 wt %; (2) Ultrafine grinding: The impurity-removed Ganoderma lucidum spore powder is ground by ultra-low temperature vibration mill to obtain partially broken Ganoderma lucidum spore powder; (3) Enzymatic hydrolysis: resuspending the partially broken spore powder in pure water, then adding neutral protease to perform enzymatic hydrolysis to obtain a Ganoderma lucidum spore powder suspension; (4) Centrifugal separation: The Ganoderma lucidum spore powder suspension is centrifuged to collect the oil phase, water phase and residue phase respectively. The oil phase is the Ganoderma lucidum spore oil.

2. The extraction method according to claim 1, characterized in that In step (1), the vacuum low-temperature drying treatment conditions are: temperature 50-60° C., time 2-4 h, vacuum degree -0.06 to -0.08 MPa.

3. The extraction method according to claim 1, characterized in that In step (2), the crushing temperature is -38 to -28°C, and the crushing time is 30 minutes to 1 hour; nitrogen protection is provided during the crushing process.

4. The extraction method according to claim 1, characterized in that In step (3), the mass ratio of spore powder to pure water during the resuspension process is 1:5-10.

5. The extraction method according to claim 1, characterized in that In step (3), the amount of neutral protease used relative to the Ganoderma lucidum spore powder is 2000-5000 U / g; and the enzymatic hydrolysis conditions are: enzymatic hydrolysis treatment at 45-50° C. for 2-4 hours.

6. The extraction method according to claim 1, characterized in that In step (4), centrifugation is performed using an automatic slag-discharging disc centrifuge; the conditions for centrifugal separation are: rotation speed 3000-9500 rpm, flow rate 1-3 t / h, and back pressure 0.2-0.3 MPa.

7. Ganoderma lucidum spore oil obtained by the extraction method according to any one of claims 1 to 6.

8. A microcapsule containing the Ganoderma lucidum spore oil according to claim 7, characterized in that: The preparation method of the microcapsule is: (1) mixing the aqueous phase obtained in the process of extracting the ganoderma spore oil with the egg yolk phospholipid protein in a vacuum emulsification tank, adjusting the pH to 7-8, stirring and mixing, and then pumping the oil phase obtained in the process of extracting the ganoderma spore oil into the vacuum emulsification tank for emulsification; (2) The emulsified material is spray-dried to obtain microcapsule powder.

9. The microcapsule according to claim 8, characterized in that In step (1), the mass ratio of the aqueous phase to the egg yolk phospholipid protein is 10:1-2; the emulsification is stirred for 15-30 minutes under the conditions of a vacuum degree of -0.06-0.08 MPa, a stirring speed of 23-38 rpm, and an emulsification speed of 2800-3600 rpm.

10. The microcapsule according to claim 8, characterized in that In step (2), the spray drying conditions are: vacuum degree -50 to -2000 Pa, inlet temperature 110 to 120° C., and outlet temperature 55 to 60° C.

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