Method for extracting tricholoma matsutake alcohol from fresh shiitake mushrooms

Abstract

The invention discloses a method for extracting tricholoma matsutake alcohol from fresh lentinus edodes, and relates to the technical field of tricholoma matsutake alcohol extraction. The method comprises the following steps: preparing fresh lentinus edodes into homogenate, adding ethanol, uniformly stirring, and performing static extraction to obtain an extraction mixture; adding water into the extracted mixture, and freezing at-80 DEG C for 12 hours to obtain an ice crystal-shaped sample; vacuumizing the ice crystal-shaped sample in an environment of-50 DEG C, maintaining the vacuum degree to be less than 20mTorr during vacuumizing, dissolving substances in cold hydrazine of a freeze dryer, and uniformly mixing to obtain tricholoma matsutake alcohol-water liquid; according to the method, static extraction and freeze drying are adopted firstly, fresh shiitake mushrooms serve as raw materials, dissolution of tricholoma matsutake alcohol in the shiitake mushrooms is fully promoted, complex extraction equipment is not needed, a large number of consumables are avoided, the extraction rate of the tricholoma matsutake alcohol is increased, and the highest concentration of the tricholoma matsutake alcohol in the substance obtained through the method is 11.8 mg / L.

Description

Technical Field

[0001] This application relates to the technical field of matsutakeol extraction, and particularly to a method for extracting matsutakeol from fresh shiitake mushrooms. Background Art

[0002] Matsutakeol, also known as mushroom alcohol (1-octen-3-ol), has been included in the food additive catalog in China. Its safety has been recognized by the FDA and FEMA, and it is widely used in various foods such as beverages, candies, ice creams, baked foods, seasonings, and tobacco flavorings, especially favored in processed food of edible fungi. Matsutakeol also has significant biological activities, as well as various health benefits such as anti-radiation and anti-gene mutation, treatment of cardiovascular diseases, reduction of blood lipids, and anti-diabetes. The conventional extraction of matsutakeol from natural products is carried out by four extraction methods: steam distillation, simultaneous distillation extraction, supercritical extraction, and solid-phase microextraction. Gas chromatography-mass spectrometry is used for its analysis and determination. All of them require extracting the sample at a relatively high temperature. The extracted product has more impurities, and it is easy to cause the volatilization and dispersion of matsutakeol, resulting in the destruction of its activity and a relatively low extraction yield.

[0003] Most of the raw materials used in the existing natural extraction of matsutakeol are matsutake mushrooms (CN202411408367.7, CN201510018428.3). Such methods have relatively high raw material costs, and the extraction yields of matsutakeol obtained are not publicly available, and it is impossible to effectively improve the extraction yield of matsutakeol.

[0004] In view of the problems existing in the current matsutakeol enrichment and extraction processes, the present invention develops a method for extracting matsutakeol from fresh shiitake mushrooms to improve the extraction rate of matsutakeol.

[0005] The information disclosed in the background art section is only intended to increase the understanding of the overall background of the present invention, and should not be regarded as an admission or any form of implication that this information constitutes the prior art already known to those of ordinary skill in the art. Summary of the Invention

[0006] This application provides a method for extracting matsutakeol from fresh shiitake mushrooms to solve the above technical problems. This method is simple to operate. Using shiitake mushrooms with a relatively low price compared to matsutake as raw materials, a large amount of matsutakeol can be obtained, which is beneficial to the extraction of matsutakeol.

[0007] This application provides a method for extracting matsutakeol from fresh shiitake mushrooms, including the following steps: After the fresh shiitake mushrooms are made into a homogenate, ethanol is added and stirred evenly, and then static extraction is carried out to obtain an extraction mixture; Water is added to the extraction mixture, and it is frozen at -80°C for 4 - 12 hours to obtain an ice crystal-like sample; The ice crystal-like sample is evacuated in an environment of -50°C, and the vacuum degree is maintained below 20 mTorr during evacuation. The substances in the cold trap of the freeze dryer are melted and mixed to obtain the ethyl palmitate aqueous solution. The volume ratio of ethanol added to the mass of the homogenate prepared from fresh mushrooms is 1:1.

[0008] Preferably, the water content of the fresh mushrooms used is greater than 85%.

[0009] Preferably, the water content of the fresh mushrooms used is 85-92%.

[0010] Preferably, the volume ratio of water added to ethanol added in the extraction mixture is 0.4-1:1.

[0011] Preferably, the static extraction is carried out at room temperature.

[0012] Preferably, the evacuation treatment time is 24-72 h.

[0013] The beneficial effects that this application can produce include: 1) The method for extracting ethyl palmitate from fresh mushrooms provided by this application. This method first uses static extraction and then freeze drying. Using fresh mushrooms as raw materials can fully promote the dissolution of ethyl palmitate in them. It does not require the use of complex extraction equipment or a large number of consumables, which helps to improve the extraction rate of ethyl palmitate. The highest concentration of ethyl palmitate in the substance obtained by this method is 11.8 mg / L, and the highest yield can reach 34 mg / kg, effectively realizing the effective extraction of ethyl palmitate from mushrooms.

[0014] 2) The method for extracting ethyl palmitate from fresh mushrooms provided by this application. This extraction method has a simple process, few steps, and short time consumption. It omits some separation and purification steps, has a short extraction cycle, and high efficiency. In this method, the use of ethanol and aqueous solution can not only effectively extract fresh mushrooms, but also enrich ethyl palmitate, inhibit the competitive precipitation of other volatile components during freeze drying, improve the absorption rate of ethyl palmitate, and thus improve the extraction yield of ethyl palmitate.

[0015] 4) The method for extracting ethyl palmitate from fresh mushrooms provided by this application. This extraction method has high selectivity, high yield, and low toxicity to ethyl palmitate, and has no adverse effects on humans and the environment. The extraction method of the present invention does not produce new three wastes and has no environmental protection problems. Description of the Drawings

[0016] Figure 1 It is the total ion current chromatogram of the ethyl palmitate standard product; Figure 2 It is the total ion current chromatogram of the substance obtained in Example 1 provided by this application; Figure 3 It is the total ion current chromatogram of the substance obtained in Example 2 provided by this application; Figure 4 Provide the total ion chromatogram of the substance obtained in Example 3 of this application; Figure 5 Provide the total ion chromatogram of the substance obtained in Example 4 of this application; Figure 6 Provide the total ion chromatogram of the substance obtained in Example 5 of this application; Detailed implementation manners The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments, but the present invention is not limited in any way. Any transformation or improvement based on the teachings of the present invention falls within the protection scope of the present invention.

[0017] Embodiment In the following embodiments, the materials and instruments used are obtained from commercial channels unless otherwise specified; the detection methods used are existing methods unless otherwise specified.

[0018] Embodiment 1 Select 500 g of fresh shiitake mushrooms (water content 92%), make a homogenate, add 500 mL of ethanol, stir well, and perform static extraction at room temperature (18°C) for 48 h to obtain an extraction mixture; Add 200 mL of water to the extraction mixture, stir well, and freeze at -80°C for 12 h to obtain an ice crystal-like sample; Quickly take out the ice crystal-like sample and put it into a freeze dryer pre-cooled to -50°C, evacuate for 24 h, and maintain the vacuum degree less than 20 mTorr; Collect the substances in the cold trap of the freeze dryer, dissolve and mix them to obtain the aqueous solution of matsutake alcohol. The total ion chromatogram of the obtained substance is as Figure 2 shown. By comparing with the total ion chromatogram of the matsutake alcohol standard product ( Figure 1 ), it can be known that the obtained substance is matsutake alcohol.

[0019] Embodiment 2 Select 1000 g of fresh shiitake mushrooms (water content 90%), make a homogenate, add 1000 mL of ethanol, stir well, and perform static extraction at room temperature (18°C) for 48 h to obtain an extraction mixture; Add 500 mL of water to the extraction mixture, stir well, and freeze at -80°C for 12 h to obtain an ice crystal-like sample; Quickly take out the ice crystal-like sample and put it into a freeze dryer pre-cooled to -50°C, evacuate for 48 h, and maintain the vacuum degree less than 20 mTorr; Collect the substances in the cold trap of the freeze dryer, dissolve and mix them to obtain the aqueous solution of matsutake alcohol.

[0020] The total ion chromatogram of the obtained substance is as Figure 3 shown. By comparing with the total ion chromatogram of the matsutake alcohol standard product ( Figure 1)By comparison, the obtained substance is matsutake alcohol.

[0021] Example 3 Select 2000 g of fresh shiitake mushrooms (with a moisture content of 88%), make a homogenate, add 2000 mL of ethanol, stir well, and perform static extraction at room temperature (18 °C) for 48 h to obtain an extraction mixture; Add 2000 mL of water to the extraction mixture, stir well, and freeze at -80 °C for 12 h to obtain an ice crystal-like sample; Quickly take out the ice crystal-like sample and put it into a freeze dryer pre-cooled to -50 °C, evacuate for 72 h, and maintain the vacuum degree less than 20 mTorr; Collect the substances in the cold trap of the freeze dryer, dissolve and mix them to obtain a matsutake alcohol aqueous solution.

[0022] The total ion current chromatogram of the obtained substance is as Figure 4 shown. By comparison with the total ion current chromatogram of the matsutake alcohol standard product ( Figure 1 ), it can be known that the obtained substance is matsutake alcohol.

[0023] Example 4 Select 1000 g of fresh shiitake mushrooms (with a moisture content of 85%), make a homogenate, add 1000 mL of ethanol, stir well, and perform static extraction at room temperature (18 °C) for 12 h to obtain an extraction mixture; Add 500 mL of water to the extraction mixture, stir well, and freeze at -80 °C for 4 h to obtain an ice crystal-like sample; Quickly take out the ice crystal-like sample and put it into a freeze dryer pre-cooled to -50 °C, evacuate for 60 h, and maintain the vacuum degree less than 20 mTorr; Collect the substances in the cold trap of the freeze dryer, dissolve and mix them to obtain a matsutake alcohol aqueous solution.

[0024] The total ion current chromatogram of the obtained substance is as Figure 5 shown. By comparison with the total ion current chromatogram of the matsutake alcohol standard product ( Figure 1 ), it can be known that the obtained substance is matsutake alcohol.

[0025] Example 5 Select 1000 g of fresh shiitake mushrooms (with a moisture content of 92%), make a homogenate, add 1000 mL of ethanol, stir well, and perform static extraction at room temperature (18 °C) for 24 h to obtain an extraction mixture; Add 500 mL of water to the extraction mixture, stir well, and freeze at -80 °C for 8 h to obtain an ice crystal-like sample; Quickly take out the ice crystal-like sample and put it into a freeze dryer pre-cooled to -50 °C, evacuate for 72 h, and maintain the vacuum degree less than 20 mTorr; Collect the substances in the cold trap of the freeze dryer, dissolve and mix them to obtain a matsutake alcohol aqueous solution.

[0026] The total ion current chromatogram of the obtained substance is as Figure 6 shown. By comparing with the total ion current chromatogram of the standard sample of matsutake alcohol ( Figure 1 ), it can be known that the obtained substance is matsutake alcohol.

[0027] Comparative Example 1 Select 1000 g of fresh shiitake mushrooms (water content 92%), slice them, and freeze at -80 °C for 4 h; quickly take them out and put them into a freeze dryer pre-cooled to -50 °C, evacuate for 48 h, and maintain the vacuum degree less than 20 mTorr; collect and mix the substances melted in the cold trap of the freeze dryer to obtain an aqueous solution.

[0028] Comparative Example 2 The difference from Example 2 is that instead of static extraction after adding ethanol solution, water with the same mass and volume as the homogenate of fresh shiitake mushrooms is added for soaking, and the rest is the same as Example 2.

[0029] For the alcohol aqueous solutions obtained in Examples 1-5 and Comparative Examples 1-2, a method for determining the content of matsutake alcohol by gas chromatography-mass spectrometry was used, and the concentration was calculated. The specific method refers to the standard (T / YNBX 075-2023, Determination of 1-octen-3-ol (matsutake alcohol) content in edible fungi - Gas chromatography-mass spectrometry method [S]). The optimized sample treatment steps are as follows: Take 100 mL of the obtained matsutake alcohol aqueous solution, add 10 mL of dichloromethane for extraction, collect the extract, add 2 g of anhydrous magnesium sulfate and shake well to remove water, and filter the supernatant through a 0.22 μm microporous filter membrane, and then measure the content of matsutake alcohol on the machine.

[0030] The active ingredients prepared by the methods of Examples 1-5 and Comparative Examples 1-2 were detected, and the detection results are shown in the following table.

[0031] Table 1 Detection results of matsutake alcohol and calculation of yield As can be seen from the above table, when the present invention processes fresh shiitake mushrooms by freeze-drying process, static extraction of fresh shiitake mushrooms with ethanol solution promotes the dissolution of matsutake alcohol, helps to improve the extraction rate and yield of matsutake alcohol, and the highest yield can reach 34 mg / kg, while matsutake alcohol cannot be extracted in Comparative Examples 1-2; the conventional freeze-drying process is superior in retaining the flavor substances of the test samples, and there is no phenomenon of overflow and dispersion of matsutake alcohol. In the present invention, the ethanol aqueous solution can not only effectively extract fresh shiitake mushrooms, but also enrich matsutake alcohol, inhibit the competition of other volatile components during freeze-drying, improve the absorption rate of matsutake alcohol, and thus improve the extraction rate. The extraction method of the present invention has the effects of high selectivity, high yield and low toxicity on matsutake alcohol, and has no adverse effects on humans and the environment.

[0032] As can be seen from the above table, by changing the form of the samples processed by this method, it is impossible to obtain matsutakeol by processing shiitake mushroom slices, or it is impossible to obtain matsutakeol without using ethanol for static extraction. This shows that all operations of this method are essential operations, and matsutakeol cannot be obtained after replacement.

[0033] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for extracting matsutakeol from fresh shiitake mushrooms, characterized in that, It includes the following steps: After making fresh shiitake mushrooms into a homogenate, add ethanol, stir evenly, and perform static extraction to obtain an extraction mixture; Add water to the extraction mixture, freeze it at -80 °C for 4 to 12 hours to obtain an ice crystal-like sample; Vacuumize the ice crystal-like sample at -50 °C. During the vacuumization, maintain the vacuum degree less than 20 mTorr. Take the substance in the cold trap of the freeze dryer, dissolve it, and mix evenly to obtain a ricinolol aqueous solution; The volume ratio of the added ethanol to the mass of the homogenate made from fresh shiitake mushrooms is 1:

1.

2. The method for extracting matsutakeol from fresh shiitake mushrooms according to claim 1, wherein The water content of the used fresh shiitake mushrooms is greater than 85%.

3. The method for extracting matsutake alcohol from fresh shiitake mushrooms according to claim 1, characterized in that, The water content of the used fresh shiitake mushrooms is 85 - 92%.

4. The method for extracting tricholomic acid from fresh shiitake mushrooms according to claim 1, characterized in that, The volume ratio of the added water to the added ethanol in the extraction mixture is 0.4 - 1:

1.

5. The method for extracting matsutake alcohol from fresh shiitake mushrooms according to claim 1, characterized in that, The static extraction is carried out at room temperature.

6. The method for extracting matsutake alcohol from fresh shiitake mushrooms according to claim 1, characterized in that, The vacuum treatment time is 24 - 72 hours.

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