A method for preparing ganoderma spore oil

Abstract

The present invention relates to a preparation method of ganoderma spore oil, which comprises the following steps: freezing with liquid nitrogen and then thawing in a warm water bath at 45°C for several cycles; then performing dual-frequency ultrasonic treatment; taking the broken-wall ganoderma spore powder and carrying out CO2 supercritical extraction, and then removing the filter residue to obtain ganoderma spore oil, and realizing the crystallization of ganoderma spore oil through the cooling rates of two gradients. The preparation method of ganoderma spore oil provided by the present invention can effectively improve the broken-wall rate of ganoderma spore powder through freezing and thawing and ultrasonic oscillation, thereby improving the yield of ganoderma spore oil. When using specific process operation parameters and combining the three, a synergistic effect can be achieved as a whole, enhancing the broken-wall rate of ganoderma spore powder, improving the yield of ganoderma spore oil and the total triterpenoid content. In addition, the total triterpenoid content of ganoderma spore oil can be further improved and the product appearance can be improved by using the winterization filtration method for ganoderma spore oil.

Description

Technical Field

[0001] The present invention relates to the technical field of Ganoderma lucidum processing, in particular to a method for preparing Ganoderma lucidum spore oil and a winterization method for Ganoderma lucidum spore oil. Background Art

[0002] Ganoderma lucidum has a medicinal history of thousands of years in China and has always been regarded as a miraculous medicinal fungus. It is commonly known as the "immortal herb" and the "resurrection herb" among the people. After Ganoderma lucidum matures, it will emit red-brown or other colored spores, which are the sexual reproductive cells of Ganoderma lucidum and are also called basidiospores. Ganoderma lucidum spores are enriched with all the genetic active ingredients of Ganoderma lucidum, and their medicinal value is much higher than that of Ganoderma lucidum fruiting bodies. Ganoderma lucidum spores have a double-layer tough chitin and peptidoglycan outer wall. When taken directly, the human body cannot digest the Ganoderma lucidum cell wall, and the effective active ingredients cannot be fully utilized. Therefore, generally, Ganoderma lucidum spores are broken walled before taking.

[0003] Ganoderma lucidum spore oil is a lipid extract of Ganoderma lucidum spores. The main components of Ganoderma lucidum spore oil include triterpenoids, steroids, and unsaturated fatty acids, and have obvious physiological activities. The oil content rate of Ganoderma lucidum spore powder varies depending on the type of Ganoderma lucidum. The oil yield of unbroken Ganoderma lucidum spore powder is only 1% - 2%, which is much lower than the actual oil content rate of Ganoderma lucidum spores. Therefore, the extraction of Ganoderma lucidum spore oil usually selects broken Ganoderma lucidum spores for oil extraction.

[0004] The wall-breaking rate of Ganoderma lucidum spore powder will affect the yield of Ganoderma lucidum spore oil and the content of active ingredients. Although there are currently various technologies that can complete the wall-breaking treatment of Ganoderma lucidum spore powder, the chemical properties of the triterpenoids contained in Ganoderma lucidum spore powder are unstable. How to maximize the guarantee that the effective ingredients of Ganoderma lucidum are not damaged during the wall-breaking process while improving the wall-breaking rate is still a problem to be solved.

[0005] In addition, due to the rich chemical composition of Ganoderma lucidum spore oil, it is easy to crystallize at low temperatures and become a white opaque flocculent product, seriously affecting the appearance of spore oil. Consumers have a low acceptance of Ganoderma lucidum spore oil capsules containing flocs. It is urgent to solve the problem of flocs generated by Ganoderma lucidum spore oil at low temperatures. Summary of the Invention

[0006] The purpose of the present invention is to provide a method for preparing Ganoderma lucidum spore oil, which can maintain a high wall-breaking rate of Ganoderma lucidum spore powder while maintaining a high oil yield and a high total triterpenoid content of spore oil.

[0007] The technical solution to achieve the purpose of the present invention is as follows: As Figure 1 shown, a method for preparing Ganoderma lucidum spore oil includes the following steps:

[0008] S1 Weigh unbroken Ganoderma lucidum spore powder, wash it with water, and dry it;

[0009] S2 is taken out after being frozen with liquid nitrogen and thawed in a warm water bath at 45°C, and the number of cycles is 1 - 5 times;

[0010] S3 The spore powder processed in step 2 is mixed with water and treated with dual-frequency ultrasound. The treatment conditions are alternating ultrasound at 20 kHz and 28 kHz;

[0011] S4 After the ultrasound ends, it is filtered, centrifuged, and dried to obtain broken-wall ganoderma spore powder;

[0012] S5 The broken-wall ganoderma spore powder is taken and extracted by CO2 supercritical extraction, and then the filter residue is removed to obtain ganoderma spore oil.

[0013] Since the broken-wall ganoderma spore powder has small particles and is easy to enter the separation kettle with gas to block the pipeline, or be entrained into the ganoderma spore oil to affect the purity of the oil, step s4 is added.

[0014] Ganoderma spore oil contains more high-melting-point fatty acids, so it is easy to crystallize at low temperatures, affecting the product appearance. Physical treatment of crystallization is the most advisable as a health product. Further, to reduce the problem of easy crystallization of ganoderma spore oil at low temperatures, the following steps are also included after step S5:

[0015] S6 The ganoderma spore oil prepared in step S5 is cooled to 15°C while stirring, and the cooling rate is 2°C / h;

[0016] S7 When the temperature is 15°C, the cooling rate is changed to 0.3°C / h until the crystallization temperature of 7°C, and the crystal cultivation time is 24 - 30 h;

[0017] S8 Filter.

[0018] By adding the above steps, not only can the problem of easy crystallization of ganoderma spore oil at low temperatures be reduced, but also the total triterpene content of ganoderma spore oil can be increased through the embodiments of the present invention.

[0019] Further, the number of freeze-thaw cycles in step S2 is 3 - 5 times.

[0020] Further, in step S3, the alternating working time of ultrasound is 3 s, the ultrasound power per unit volume is 40 W / L, the initial temperature is 25°C, and the ultrasound treatment time is 15 - 20 min.

[0021] Further, in step S5, the pressure of supercritical extraction is 35 MPa, the time is 3 h, and the temperature is 48°C.

[0022] Further, in step S3, the weight ratio of spore powder to water is 1:15.

[0023] Further, the number of freeze-thaw cycles in step S2 is 3 times; the ultrasound treatment time in step S3 is 15 - 20 min.

[0024] The present invention also provides a method for winterization of ganoderma spore oil, which is characterized by comprising the following steps:

[0025] S1 Take the ganoderma spore oil and cool it to 15°C while stirring, with a cooling rate of 2°C / h;

[0026] S2 When the temperature reaches 15°C, change the cooling rate to 0.3°C / h until the crystallization temperature of 7°C is reached, and the crystal cultivation time is 24 - 30 h;

[0027] S3 Filter.

[0028] Compared with the prior art, the significant advantages of the present invention are as follows: The preparation method of ganoderma spore oil provided by the present invention can effectively improve the wall-breaking rate of ganoderma spore powder through freeze-thawing and ultrasonic oscillation, thereby increasing the yield of ganoderma spore oil. By using specific process operation parameters and combining the three, a synergistic effect can be achieved as a whole, enhancing the wall-breaking rate of ganoderma spore powder, increasing the yield of ganoderma spore oil and the total triterpene content. In addition, the method of winterization filtration of ganoderma spore oil can further increase the total triterpene content of ganoderma spore oil and improve the product appearance. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is a schematic flow chart of the preparation method of ganoderma spore oil of the present invention.

[0030] Figure 2 is a schematic diagram of the comparison of experimental result data of Examples 3 and 5 and Comparative Examples 1 and 2 of the present invention.

[0031] Figure 3 is a schematic diagram of the comparison of experimental result data of Example 3 and Comparative Examples 3 and 4 of the present invention.

[0032] Figure 4 is a schematic diagram of the comparison of experimental result data of Examples 1, 3, and 4 of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0033] According to the following embodiments, the present invention can be better understood. It should be noted that the experimental methods without specific conditions indicated in the following embodiments are all conventional methods.

[0034] Preparation of Ganoderma Spore Oil

[0035] Example 1

[0036] The preparation method of ganoderma spore oil is carried out according to the following steps:

[0037] 1. Weigh the unbroken ganoderma spore powder, wash it with water, and then dry it;

[0038] 2. Pack the Ganoderma lucidum spore powder with polyester cloth, freeze it quickly with liquid nitrogen for 1 minute, then quickly take it out and thaw it in a warm water bath (45°C) for 15 minutes, and repeat this cycle 3 times;

[0039] 3. Take the spore powder treated in step 2 and mix it with water and put it into a dual-frequency focused ultrasonic equipment for treatment. The weight ratio of spore powder to water is 1:15. The treatment conditions are 20kHz / 28kHz alternating ultrasound, ultrasonic alternating working time is 3s, unit volume ultrasonic power is 40W / L, initial temperature is 25°C, and ultrasonic treatment is 10min;

[0040] 4. After the ultrasound is finished, filter, centrifuge and dry to obtain the broken spore powder of Ganoderma lucidum;

[0041] 5. The broken spore powder of Ganoderma lucidum is subjected to supercritical CO2 extraction (pressure of 35 MPa, time of 3 h, temperature of 48° C.), and then the filter residue is removed to obtain Ganoderma lucidum spore oil.

[0042] Example 2

[0043] Except that the number of freeze-thaw cycles in step 2 was 5, the rest of the preparation method was carried out according to Example 1.

[0044] Example 3

[0045] Except for step 3 in which the ultrasonic treatment time was 15 min, the rest of the preparation method was carried out according to Example 1.

[0046] Example 4

[0047] Except for step 3 in which the ultrasonic treatment time was 20 min, the rest of the preparation method was carried out according to Example 1.

[0048] Example 5

[0049] The preparation method was carried out according to Example 1, except that the number of freeze-thaw cycles in step 2 was 5 and the ultrasonic treatment time in step 3 was 15 min.

[0050] Example 6

[0051] Except that the number of freeze-thaw cycles in step 2 was 5 and the ultrasonic treatment time in step 3 was 20 min, the rest of the preparation method was carried out according to Example 1.

[0052] Comparative Example 1

[0053] The preparation method was carried out according to Example 1, except that the number of freeze-thaw cycles in step 2 was 1 and the ultrasonic treatment time in step 3 was 15 min.

[0054] Comparative Example 2

[0055] The preparation method was carried out according to Example 1, except that the number of freeze-thaw cycles in step 2 was 7 and the ultrasonic treatment time in step 3 was 15 min.

[0056] Comparative Example 3

[0057] Except for step 3 in which the spore powder is mixed with water and placed in a single-frequency focused ultrasonic device and ultrasonically treated at 20 kHz for 15 min, the rest of the preparation method is carried out according to Example 1.

[0058] Comparative Example 4

[0059] The preparation method is carried out according to Example 1 except that in step 3, the spore powder is mixed with water, put into a single-frequency focused ultrasonic device, and ultrasonically treated at 28 kHz for 15 minutes.

[0060] Comparative Example 5

[0061] Except for step 3 in which the spore powder is mixed with water and placed in a single-frequency focused ultrasonic device and ultrasonically treated at 20 kHz for 20 min, the rest of the preparation method is carried out according to Example 1.

[0062] Comparative Example 6

[0063] Except for step 3 in which the spore powder is mixed with water and placed in a single-frequency focused ultrasonic device and ultrasonically treated at 28 kHz for 20 min, the rest of the preparation method is carried out according to Example 1.

[0064] The performance tests of Examples 1-5 and Comparative Examples 1-6 were carried out respectively, as follows:

[0065] Testing method for the cell-breaking rate: The cell-breaking spore powder of Ganoderma lucidum after step 4 of the embodiment and the comparative example is placed under a microscope, and the cell-breaking rate is calculated by the hemocytometer method.

[0066] Test method for the content of total triterpenoid compounds in Ganoderma lucidum: The Ganoderma lucidum spore oil prepared in the examples and comparative examples was measured by colorimetry, the total triterpenoid content was measured by vanillin-glacial acetic acid-perchloric acid method, glucose and oleanolic acid were used as controls, the absorbance of the sample was measured by spectrophotometry at 548nm and the corresponding concentration was calculated according to the standard curve. (Specific reference is made to the method specified in "Determination of total triterpenoids in health foods" in the "Technical Guidelines for Inspection and Evaluation of Physical, Chemical and Hygiene Indicators of Health Foods" (2020 edition).

[0067] Table 1

[0068]

[0069] By comparing the preparation methods of Examples 3 and 5 with those of Comparative Examples 1 and 2, and the performance test results, it can be seen that Figure 2As shown in the figure, when the ultrasonic frequency and ultrasonic time are the same, the freezing and thawing cycle times increase, the wall breaking rate of Ganoderma lucidum spore powder is higher, but the fewer (Comparative Example 1) or more (Comparative Example 2) cycles do not increase the yield of Ganoderma lucidum spore oil, and the content of total triterpenes is not high. This may be due to the excessive number of cold and hot cycles in Comparative Example 2, which destroys the

[0070] By comparing the preparation methods of Example 3 with those of Comparative Examples 3 and 4 and the performance test results, it can be seen that Figure 3 As shown: with the same number of freeze-thaw cycles and the same ultrasonic treatment time, the effect of dual-frequency ultrasound is far superior to that of single-frequency ultrasound. This is because after the freeze-thaw cycle, gaps appear in the spores, and the staggered ultrasonic oscillation frequency is more conducive to breaking the spore wall, increasing the spore oil yield and greatly protecting the content of total triterpenes.

[0071] By comparing the preparation methods of Example 4 with those of Comparative Examples 5 and 6 and the performance test results, it can be seen that with the same number of freeze-thaw cycles and the same ultrasonic treatment time, the dual-frequency ultrasonic effect is far superior to the single-frequency ultrasonic effect. This is because after the freeze-thaw cycle, gaps are generated in the spores, and staggered ultrasonic oscillation frequencies are more conducive to breaking the spore wall, improving the spore oil yield and greatly protecting the content of total triterpenes.

[0072] By comparing the preparation methods of Examples 1, 3, and 4 with the performance test results, and comparing the preparation methods of Examples 2, 5, and 6 with the performance test results, it can be seen that: with the same number of freeze-thaw cycles, the same dual-frequency ultrasonic treatment, and appropriate ultrasonic treatment time, the wall-breaking rate of spore powder can be improved, but the dual-frequency treatment will generate a lot of heat, which will cause the spore oil to volatilize.

[0073] It can be seen that in the preparation method of Ganoderma lucidum spore oil provided by the present invention, the number of freezing cycles, ultrasonic vibration frequency and ultrasonic treatment time are combined, and specific process operation parameters are used to achieve a synergistic effect as a whole, and ultimately obtain a high Ganoderma lucidum spore oil extraction rate and a higher total triterpene content.

[0074] Winterization preparation of Ganoderma lucidum spore oil

[0075] Example 7

[0076] The Ganoderma lucidum spore oil prepared in Example 3 was winterized, and the specific operation steps were as follows:

[0077] 1. Take the Ganoderma lucidum spore oil (original temperature is 45° C.) prepared in Example 3 and cool it to 15° C. while stirring, at a cooling rate of 2° C. / h;

[0078] 2. Change the cooling rate to 0.3℃ / h, to the crystallization temperature of 7℃, and the crystal growth time is 24h;

[0079] 3. Filter the flocs to obtain transparent Ganoderma lucidum spore liquid oil.

[0080] The obtained transparent Ganoderma lucidum spore liquid oil was subjected to a freezing transparency experiment at 0°C and could remain clear and transparent for 8 hours.

[0081] Example 8

[0082] The Ganoderma lucidum spore oil prepared in Example 3 was winterized, and the specific operation steps were as follows:

[0083] 1. Take the Ganoderma lucidum spore oil (original temperature is 45° C.) prepared in Example 3 and cool it to 15° C. while stirring, at a cooling rate of 2° C. / h;

[0084] 2. Change the cooling rate to 0.3℃ / h, to the crystallization temperature of 7℃, and the crystal growth time is 30h;

[0085] 3. Filter the flocs to obtain transparent Ganoderma lucidum spore liquid oil.

[0086] The obtained transparent Ganoderma lucidum spore liquid oil was subjected to a freezing transparency experiment at 0°C and could remain clear and transparent for 8.5 hours.

[0087] The Ganoderma lucidum spore oil prepared in Example 3 and the Ganoderma lucidum spore liquid oils in Examples 7 and 8 were measured for total triterpenoid content according to the method specified in "Determination of total triterpenoids in health foods" in the "Technical Guidelines for Inspection and Evaluation of Physical, Chemical and Hygiene Indicators of Health Foods" (2020 Edition). The results are shown in Table 2

[0088] Table 2

[0089]

[0090] By comparing the total triterpene content in Examples 3, 7, and 8, it was found that after winterization, the active ingredient content of the spore oil was slightly increased. This may be because a part of the solid oil was lost during winterization, and the active ingredients were more concentrated in the liquid oil, so the active ingredient content was slightly increased.

[0091] The preparation method of ganoderma lucidum spore oil provided by the present invention can effectively improve the wall-breaking rate of ganoderma lucidum spore powder through freezing and thawing, and ultrasonic oscillation, thereby improving the yield of ganoderma lucidum spore oil. When using specific process operation parameters, the three are combined to play a synergistic role as a whole, enhance the wall-breaking rate of ganoderma lucidum spore powder, and improve the yield and total triterpene content of ganoderma lucidum spore oil. In addition, the method of winterizing and filtering ganoderma lucidum spore oil can further improve the total triterpene content of ganoderma lucidum spore oil and improve the product appearance, which is conducive to large-scale industrial preparation method.

[0092] It should be understood that although this specification is described in terms of embodiments, not every embodiment contains only one independent technical solution. This narrative manner of the specification is only for clarity. Those skilled in the art should regard the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for preparing ganoderma spore oil, characterized in that: It is prepared by the following steps: S1 Weigh the unbroken ganoderma lucidum spore powder, wash it with water, and dry it; S2 Freeze it with liquid nitrogen and then take it out and thaw it in a warm water bath at 40 °C to 45 °C. The number of cycles is 3 to 5 times; S3 Take the spore powder treated in step 2, mix it with water, and use dual-frequency ultrasonic treatment. The treatment conditions are alternating ultrasonic waves at 20 kHz and 28 kHz; S4 After the ultrasonic treatment is completed, filter, centrifuge, and dry to obtain broken ganoderma lucidum spore powder; S5 Take the broken ganoderma lucidum spore powder and carry out CO2 supercritical extraction, and then remove the filter residue to obtain ganoderma lucidum spore oil; In step S3, the alternating working time of ultrasonic waves is 3 s, the ultrasonic power per unit volume is 40 W / L, the initial temperature is 25 °C, and the ultrasonic treatment time is 15 to 20 min.

2. The preparation method of ganoderma spore oil according to claim 1, characterized in that: After step S5, the following steps are also included: S6 Cool the ganoderma lucidum spore oil prepared in step S5 to 15 °C while stirring, and the cooling rate is 2 °C / h; S7 When the temperature is 15 °C, change the cooling rate to 0.3 °C / h until the crystallization temperature is 7 °C, and the crystal cultivation time is 24 to 30 h; S8 Filter.

3. The preparation method of ganoderma spore oil according to claim 1, characterized in that: In step S5, the pressure of supercritical extraction is 35 MPa, the time is 3 h, and the temperature is 48 °C.

4. The preparation method of ganoderma spore oil according to claim 1, characterized in that: In step S3, the weight ratio of spore powder to water is 1:

15.

5. The preparation method of ganoderma spore oil according to claim 1, characterized in that: The number of freeze-thaw cycles in step S2 is 3 times; the ultrasonic treatment time in step S3 is 15 to 20 min.

6. A method for winterizing ganoderma spore oil, characterized in that: It includes the following steps: S1 Take the ganoderma lucidum spore oil prepared by the method for preparing ganoderma lucidum spore oil according to any one of claims 1, 3, 4, or 5, cool it to 15 °C while stirring, and the cooling rate is 2 °C / h; S2 When the temperature is 15 °C, change the cooling rate to 0.3 °C / h until the crystallization temperature is 7 °C, and the crystal cultivation time is 24 to 30 h; S3 Filter.

Images