A sauce-flavored distiller's grains extract, a preparation method thereof, and application thereof in anti-inflammatory electronic cigarette oil

Abstract

The present invention provides a sauce-flavored wine lees extract, a preparation method thereof, and an application thereof in anti-inflammatory electronic cigarette oil, and relates to the technical field of industrial waste treatment. The preparation method of the sauce-flavored wine lees extract of the present invention comprises: hydrolyzing the sauce-flavored wine lees with a composite enzyme, filtering, centrifuging the filtrate to obtain a supernatant, extracting the extract with supercritical carbon dioxide, and removing impurities from the extract to obtain the sauce-flavored wine lees extract. The sauce-flavored wine lees extract of the present invention has a typical and rich ester aroma, which is elegant and mellow, and can last for a long time. Experimental results show that the sauce-flavored wine lees extract of the present invention can significantly inhibit the secretion of the inflammatory factor IL-6 in mouse mononuclear macrophages RAW264.7, which shows that the sauce-flavored wine lees extract of the present invention has anti-inflammatory effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of industrial waste treatment, and in particular relates to a sauce-flavored distiller's grains extract, a preparation method thereof, and application thereof in anti-inflammatory electronic cigarette oil. Background Art

[0002] Distillers' grains are the residue left after brewing rice, wheat, sorghum, and other liquors. Currently, liquor companies generate large quantities of waste grains. Direct storage of these waste grains can easily lead to environmental pollution and damage. Conventional methods for distiller's grains involve feeding them directly to feed mills or breeding operations. However, large-scale breeding operations rarely exist in a single region to consume large quantities of wet distiller's grains. Transporting these grains to other locations is not only costly but also time-consuming. Due to the effects of residual bacteria in the grains, these wet distiller's grains have a short shelf life and, if not used within 2-3 days, they deteriorate, generating significant amounts of solid waste that cannot be reused. Therefore, the rational utilization of distiller's grains is an urgent issue that needs to be addressed.

[0003] Currently, there are various ways to comprehensively utilize distiller's grains waste, including using them as energy, fertilizer, feed, and materials. However, there is no record of using Maotai-flavor distiller's grains to prepare e-cigarette oil. Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a sauce-flavored distiller's grains extract, a preparation method thereof, and an application in anti-inflammatory electronic cigarette oil. The sauce-flavored distiller's grains extract has anti-inflammatory effects and provides a new approach for the rational utilization of distiller's grains.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides a preparation method of a sauce-flavored wine lees extract, comprising: hydrolyzing the sauce-flavored wine lees with a composite enzyme, filtering, centrifuging the filtrate to obtain a supernatant, extracting the supernatant with supercritical carbon dioxide, and removing impurities from the extract to obtain the sauce-flavored wine lees extract; the composite enzyme comprises composite cellulase and hemicellulase, and the mass ratio of the composite cellulase to the hemicellulase is 1:1-2.

[0007] Preferably, the enzyme activity of the complex enzyme is (3-5)×10 4 U / g.

[0008] Preferably, the added amount of the complex enzyme is 1% to 2% of the mass of the sauce-flavor distiller's grains.

[0009] Preferably, the enzymatic hydrolysis conditions are: pH 4.5-6.5, temperature 45-55° C., and time 5-7 h.

[0010] Preferably, the centrifugal speed is 9000-11000 r / min, and the time is 15-25 min.

[0011] Preferably, the extraction parameters are: the pressure of the extraction kettle is 25-35 MPa, the temperature is 45-50°C, and the frequency of the carbon dioxide pump is 20-25 r / min; the pressure of the separation kettle I is 5-10 MPa, the temperature is 45-50°C, and the charging factor is 0.3-0.7.

[0012] Preferably, the impurity removal comprises dissolving the extract with a 90% to 98% ethanol solution by volume, centrifuging and distilling to obtain the sauce-flavored wine lees extract.

[0013] The present invention also provides a sauce-flavored distiller's grains extract obtained by the above preparation method.

[0014] The present invention also provides the use of the above-mentioned Maotai-flavor distiller's grains extract in anti-inflammatory electronic cigarette oil.

[0015] Preferably, the added mass percentage of the sauce-flavor distiller's grains extract in the anti-inflammatory electronic cigarette oil is 0.1-10%.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention provides a method for preparing a Maotai-flavor distiller's grains extract, comprising: enzymatically hydrolyzing the Maotai-flavor distiller's grains with a complex enzyme, filtering, centrifuging the filtrate to obtain a supernatant, extracting the supernatant with supercritical carbon dioxide, and removing impurities from the extract to obtain the Maotai-flavor distiller's grains extract. The Maotai-flavor distiller's grains extract of the present invention comprises characteristic components selected from the group consisting of lactic acid, glycerol, propylene glycol, butyl acetate, propionic acid, butanol, and benzyl alcohol. It contains no ethanol, has a high viscosity, and possesses a typical, rich, elegant, and mellow ester aroma that lasts for a long time. There are no similar products on the market.

[0018] The experimental results show that the sauce-flavored distiller's grains extract of the present invention can significantly inhibit the secretion of the inflammatory factor IL-6 in mouse mononuclear macrophage RAW264.7, which shows that the sauce-flavored distiller's grains extract of the present invention has anti-inflammatory effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a chromatogram of the vitamin C content of the Maotai-flavor distiller's grains extract from Example 1 diluted 10 times;

[0020] Figure 2 This is a chromatogram of the vitamin C content of the Maotai-flavor wine lees extract (sterilized at high temperature and high pressure) diluted 10 times in Example 1;

[0021] Figure 3 This is a chromatogram of the vitamin C content of the Maotai-flavor distiller's grains extract of Example 1 diluted 40 times;

[0022] Figure 4 This is a chromatogram of the vitamin C content of the Maotai-flavor wine lees extract (sterilized at high temperature and high pressure) diluted 40 times in Example 1;

[0023] Figure 5 This is a chromatogram of the vitamin E content of the Maotai-flavor distiller's grains extract of Example 1 diluted 10 times;

[0024] Figure 6 This is a chromatogram of the vitamin E content of the Maotai-flavor wine lees extract (sterilized at high temperature and high pressure) diluted 10 times in Example 1;

[0025] Figure 7 This is a chromatogram of the vitamin E content of the Maotai-flavor distiller's grains extract of Example 1 diluted 40 times;

[0026] Figure 8 This is a chromatogram of the vitamin E content of the Maotai-flavor wine lees extract (sterilized at high temperature and high pressure) diluted 40 times in Example 1;

[0027] Figure 9 Example 1: Diagram of chorioallantoic membrane blood vessels after 10-fold dilution of wine lees extract (not sterilized by high temperature and high pressure);

[0028] Figure 10 Example 1: Diagram of chorioallantoic membrane blood vessels diluted 10 times with wine lees extract (sterilized at high temperature and high pressure);

[0029] Figure 11 Example 1: Diagram of chorioallantoic membrane blood vessels after 40-fold dilution of vinasse extract (not sterilized by high temperature and high pressure);

[0030] Figure 12 Example 1: Distillers' lees extract (sterilized at high temperature and high pressure) diluted 40 times

[0031] Figure 13 The inhibition rates of different concentrations of Maotai-flavor wine lees extract on IL-6. DETAILED DESCRIPTION

[0032] The invention provides a preparation method of a sauce-flavored wine lees extract, comprising: hydrolyzing the sauce-flavored wine lees with a composite enzyme, filtering, centrifuging the filtrate to obtain a supernatant, extracting the supernatant with supercritical carbon dioxide, and removing impurities from the extract to obtain the sauce-flavored wine lees extract; the composite enzyme comprises composite cellulase and hemicellulase, and the mass ratio of the composite cellulase to the hemicellulase is 1:1-2.

[0033] The sauce-flavored wine lees of the present invention meet the raw material standards (fresh discarded lees after the seventh round of wine production, with a moisture content of no more than 60% and no less than 20%, no unpleasant odor, and a normal brown-black appearance). The sauce-flavored wine lees are preferably artificially loosened and do not release water when held by hand.

[0034] The enzyme activity of the complex enzyme of the present invention is preferably (3-5)×10 4 U / g, more preferably 4×10 4 U / g; the addition amount of the complex enzyme is preferably 1% to 2% of the mass of the sauce-flavor wine lees, more preferably 1.5%; the conditions for the enzymatic hydrolysis are preferably: pH value 4.5 to 6.5, temperature 45 to 55°C, and time 5-7h, more preferably: pH value 5, temperature 40°C, and time 6h; the centrifugal speed is preferably 9000 to 11000 r / min, more preferably 10000 r / min, and the time is preferably 15 to 25min, more preferably 20min; the pH value is preferably adjusted by sodium hydroxide.

[0035] The extraction parameters of the present invention are preferably: the pressure of the extraction kettle is 25-35MPa, the temperature is 45-50°C, and the frequency of the carbon dioxide pump is 20-25r / min; the pressure of the separation kettle I is 5-10MPa, the temperature is 45-50°C, and the charging factor is 0.3-0.7. The extraction parameters are more preferably: the pressure of the extraction kettle is 30MPa, the temperature is 40°C, the frequency of the carbon dioxide pump is 22r / min; the pressure of the separation kettle I is 8MPa, the temperature is 47°C, and the charging factor is 0.5. The present invention performs supercritical carbon dioxide extraction on the supernatant after enzymatic hydrolysis, which can obtain highly active extracts (phenolic substances and organic acids) in sauce-flavored wine lees, improve the antioxidant activity of the extract, remove fermentation inhibitors (acetic acid, lactic acid, 5-hydroxymethylfurfural (HMF), phenol and furfural) in sauce-flavored wine lees, and improve the safety of the extract.

[0036] The impurity removal of the present invention preferably includes dissolving the extract with a 90% to 98% ethanol solution by volume, centrifuging and distilling to obtain the sauce-flavored wine lees extract.

[0037] The characteristic components of the sauce-flavored distiller's grains extract of the present invention are several substances selected from lactic acid, glycerol, propylene glycol, butyl acetate, propionic acid, butanol, and benzyl alcohol. The product does not contain ethanol, has high viscosity, and is light yellow or yellow. The product has a typical and rich ester aroma, which is elegant and mellow and can last for a long time. There is no similar product on the market.

[0038] The present invention also provides a sauce-flavored distiller's grains extract obtained by the above preparation method.

[0039] The present invention also provides the use of the above-mentioned Maotai-flavor distiller's grains extract in anti-inflammatory electronic cigarette oil.

[0040] The present invention uses the sauce-flavored distiller's grains extract for inflammatory-stimulated mouse mononuclear macrophages RAW264.7. Experimental results show that the sauce-flavored distiller's grains extract of the present invention can significantly inhibit the secretion of inflammatory factor IL-6 in the inflammatory-stimulated mouse mononuclear macrophages RAW264.7. It can be seen that the sauce-flavored distiller's grains extract of the present invention has anti-inflammatory effect.

[0041] The added mass percentage of the Maotai-flavor distiller's grains extract in the anti-inflammatory electronic cigarette oil is preferably 0.1-10%, more preferably 5%.

[0042] The present invention has no special limitation on the source of the raw materials, and conventional commercial products in the field can be used.

[0043] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0044] Example 1

[0045] Preparation of Maotai-flavor Distiller's Grains Extract

[0046] 1. 500 kg of Maotai-flavor wine lees that had been loosened manually and did not produce water when held by hand were adjusted to pH 5 with sodium hydroxide and mixed with 7.5 kg of complex enzyme (total enzyme activity of 4 × 10 4 U / g, the mass ratio of complex cellulase and hemicellulase was 1:1.5) in a drum-type solid-state stirring tank, and enzymatic hydrolysis was carried out at 50°C for 6 h. After filtration, the supernatant was collected by centrifugation at 10000 r / min for 20 min.

[0047] 2. The supernatant was added to the supercritical carbon dioxide equipment for extraction. The extraction conditions were: the pressure of the extraction kettle was 30 MPa, the temperature was 40°C, and the frequency of the carbon dioxide pump was 22 r / min; the pressure of the separation kettle I was 8 MPa, the temperature was 47°C, and the charging factor was 0.5;

[0048] 3. Collect the extract and repeatedly dissolve, centrifuge and distill with 95% ethanol to remove impurities. After cleaning and sterilization, seal the fresh-keeping bag and store it at -20℃ after vacuuming.

[0049] Example 2

[0050] Preparation of Maotai-flavor Distiller's Grains Extract

[0051] 1. 500 kg of Maotai-flavor wine lees that had been loosened manually and did not produce water when held by hand were adjusted to pH 4.5 with sodium hydroxide and mixed with 5 kg of complex enzyme (total enzyme activity of 3×10 4 U / g, the mass ratio of complex cellulase and hemicellulase was 1:1) in a drum-type solid-state stirring tank, and enzymatic hydrolysis was carried out at 55°C for 5 h. After filtration, the supernatant was collected by centrifugation at 9000 r / min for 25 min.

[0052] 2. The supernatant was added to the supercritical carbon dioxide equipment for extraction. The extraction conditions were: the pressure of the extraction kettle was 25 MPa, the temperature was 50°C, and the frequency of the carbon dioxide pump was 20 r / min; the pressure of the separation kettle I was 5 MPa, the temperature was 50°C, and the charging factor was 0.3;

[0053] 3. Collect the extract and repeatedly dissolve it in 90% ethanol, centrifuge it, and distill it to remove impurities. After cleaning and sterilization, seal the fresh-keeping bag and store it at -20℃ after vacuuming.

[0054] Example 3

[0055] Preparation of Maotai-flavor Distiller's Grains Extract

[0056] 1. 500 kg of Maotai-flavor wine lees that had been loosened manually and did not produce water when held by hand were adjusted to pH 6.5 with sodium hydroxide and mixed with 10 kg of complex enzyme (total enzyme activity of 5×10 4 U / g, the mass ratio of complex cellulase and hemicellulase was 1:2) in a drum-type solid-state stirring tank, and enzymatic hydrolysis was carried out at 45°C for 7 h. After filtration, the supernatant was collected by centrifugation at 11000 r / min for 15 min.

[0057] 2. The supernatant was added to the supercritical carbon dioxide equipment for extraction. The extraction conditions were: the pressure of the extraction kettle was 35 MPa, the temperature was 40°C, and the frequency of the carbon dioxide pump was 25 r / min; the pressure of the separation kettle I was 10 MPa, the temperature was 45°C, and the charging factor was 0.7;

[0058] 3. Collect the extract and repeatedly dissolve, centrifuge and distill with 98% ethanol to remove impurities. After cleaning and sterilization, seal the fresh-keeping bag and store it at -20℃ after vacuuming.

[0059] Example 4

[0060] 2 g of the sauce-flavored distiller's grains extract prepared in Example 1 was mixed with 98 g of electronic base cigarette oil (the mass ratio of propylene glycol to glycerol was 78:20) to obtain anti-inflammatory electronic cigarette oil.

[0061] Example 5

[0062] 2 g of the sauce-flavored distiller's grains extract prepared in Example 2 was mixed with 98 g of electronic base cigarette oil (the mass ratio of propylene glycol to glycerol was 80:18) to obtain anti-inflammatory electronic cigarette oil.

[0063] Example 6

[0064] 2 g of the sauce-flavored distiller's grains extract prepared in Example 2 was mixed with 98 g of electronic base cigarette oil (the mass ratio of propylene glycol to glycerol was 75:23) to obtain anti-inflammatory electronic cigarette oil.

[0065] Example 7

[0066] Contents of vitamin C, vitamin E, tannin and solid content in vinasse extract

[0067] The sauce-flavored wine lees extract obtained in Example 1 was sterilized at high temperature and high pressure (steam pressure 205.8 kPa (2.1 kg / cm 2 ), the temperature reaches above 132°C and is maintained for 10 minutes).

[0068] The sauce-flavored wine lees extract of Example 1 and the sauce-flavored wine lees extract sterilized at high temperature and high pressure were diluted 10 times and 40 times respectively.

[0069] Method for diluting the vinasse extract: Mix the vinasse extract of Example 1 with distilled water to obtain a vinasse extract dilution solution with a corresponding dilution multiple.

[0070] 1. Determination of vitamin C content

[0071] A Shim-pack GIST C(18) column (4.6 mm × 250 mm, 5 μm) was used; the mobile phase consisted of B: acetonitrile and A: 0.1 mol / l potassium dihydrogen phosphate aqueous solution, with a ratio of A:B = 80:20; the flow rate was 1 mL min -1 ; The detection wavelength was 246 nm, the injection volume was 10 μL, and the column temperature was 40°C.

[0072] The 10-fold and 40-fold diluted concentrations of the Maotai-flavor distiller's grains extract from Example 1 and the Maotai-flavor distiller's grains extract sterilized with high temperature and high pressure were filtered through a 0.45 μm filter membrane for testing. The method standard is from "HPLC Determination of Vitamin C in Vitamin C Glucose Injection," Chinese Pharmaceutical Standards, 2007, Vol. 8, No. 2.

[0073] The peak time of the chromatogram of vitamin C standard is 2.403min.

[0074] The chromatograms of the 10-fold and 40-fold diluted sauce-flavored wine lees extracts of Example 1 and the sauce-flavored wine lees extracts sterilized at high temperature and high pressure are shown in FIG. Figures 1-4 The vitamin C content was calculated by standard curve and shown in Table 1.

[0075] Table 1 Vitamin C content of vinasse extract

[0076]

[0077]

[0078] 2. Determination of vitamin E content

[0079] A Shim-pack GIST C(18) column (4.6 mm × 250 mm, 5 μm) was used; the mobile phase was B:methanol and A:water, with a ratio of A:B = 50:50; the flow rate was 1 mL min -1 ; The detection wavelength was 275 nm, the injection volume was 10 μL, and the column temperature was 30°C.

[0080] The 10-fold and 40-fold diluted concentrations of the sauce-flavored wine lees extract of Example 1 and the high-temperature and high-pressure sterilized sauce-flavored wine lees extract were filtered through a 0.45 μm filter membrane for testing. The method standard is from "HPLC Method for Determination of Vitamin E Content in Vitamin E Tablets", Journal of Chengdu Medical College.

[0081] The peak time of the chromatogram of vitamin E standard is 5.809min.

[0082] The chromatograms of the 10-fold and 40-fold diluted sauce-flavored wine lees extracts of Example 1 and the sauce-flavored wine lees extracts sterilized at high temperature and high pressure are shown in FIG. Figure 5-Figure 8 The vitamin E content was calculated using the standard curve and is shown in Table 2.

[0083] Table 2 Vitamin E content of vinasse extract

[0084]

[0085] 3. Determination of tannin content

[0086] The Maotai-flavor wine lees extract of Example 1 diluted 10 times and 40 times and the Maotai-flavor wine lees extract sterilized at high temperature and high pressure were filtered through a 0.45 μm filter membrane for testing. The experimental method was operated according to the UV spectrophotometer QL-SOP-YF-3-30 method.

[0087] Tannin detection standard curve, good linearity, Y = 0.001X-0.0103, R 2 =0.9984.

[0088] The tannin contents of the 10-fold and 40-fold diluted sauce-flavored vinasse extracts of Example 1 and the sauce-flavored vinasse extracts sterilized at high temperature and high pressure are shown in Table 3.

[0089] Table 3 Tannin content of vinasse extract

[0090]

[0091]

[0092] 4. Solid content determination

[0093] The Maotai-flavor vinasse extract from Example 1, diluted 10-fold and 40-fold, and the Maotai-flavor vinasse extract sterilized at high temperature and pressure, were filtered through a 0.45 μm filter membrane for testing. A specific volume of the test sample was accurately measured and placed in a suitable glass weighing bottle that had been dried to a constant weight. The bottle was then oven-dried at 50°C to a constant weight. The experimental procedure was performed according to QL-SOP-YF-3-20. Specific results are shown in Table 4.

[0094] Table 4 Solid content of vinasse extract

[0095]

[0096] Example 8

[0097] Safety testing of vinasse extract

[0098] 1. HET-CAM test

[0099] The 10-fold and 40-fold diluted Maotai-flavor wine lees extracts of Example 1 and the Maotai-flavor wine lees extracts sterilized with high temperature and high pressure were filtered through a 0.45 μm filter membrane for testing, and the safety and irritation of the Maotai-flavor wine lees extracts with different treatments were compared.

[0100] Dilution of the vinasse extract: the vinasse extract of Example 1 was mixed with distilled water to obtain a vinasse extract dilution solution with a corresponding dilution multiple.

[0101] Six intact, nine-day-old chick embryos were exposed to a specific amount of the test substance directly on the chorioallantoic membrane. After three minutes, changes in the chorioallantoic membrane blood vessels were observed. The experiment was conducted according to the "Chicken Chorioallantoic Membrane Assay (CAMVA)" for eye irritation / corrosion, with both positive and negative controls. The positive control was 0.1 mol / L sodium hydroxide, and the negative control was 0.9% sodium chloride solution.

[0102] Depend on Figures 9-12 It can be seen that both liquors, diluted 10-fold and 40-fold respectively, showed no noticeable irritation on the allantoic membrane surface before and after application, with an ES score of 0, less than 12, indicating no irritation. The experimental data analysis showed that both samples were non-irritating.

[0103] 2. Human patch test

[0104] The 10-fold and 40-fold diluted Maotai-flavor wine lees extracts of Example 1 and the Maotai-flavor wine lees extracts sterilized with high temperature and high pressure were filtered through a 0.45 μm filter membrane for testing to compare the potential for adverse reactions to human skin caused by the Maotai-flavor wine lees extracts treated with different methods.

[0105] The selected area shall not exceed 50mm 2Qualified spot chamber equipment with a depth of approximately 1mm was used. The test substance was placed in the spot chamber, approximately 0.020g-0.025g (solid or semi-solid) or 0.020mL-0.025mL (liquid). The spot chamber containing the test substance was applied to the subject's forearm for 24 hours. Afterwards, the spot chamber was removed and the skin reaction was observed and the results recorded. The experimental method was carried out in accordance with QL-SOP-YF-3-04. The blank control was distilled water. Specific results are shown in Table 5.

[0106] Table 5 Human patch test results of vinasse extract

[0107]

[0108] The human patch test examines the possibility that the test substance will cause adverse reactions on human skin. If more than two people experience a Grade 2 adverse skin reaction, or if any one person experiences a Grade 3 or higher adverse skin reaction, the test substance is considered to have an adverse reaction on the human body. As shown in Table 5, all subjects participating in this experiment experienced a Grade 0 reaction. Therefore, none of the four samples caused adverse reactions on human skin. The experimental results in Table 5 indicate that the lees extract from Example 1 of the present invention is highly safe for human use.

[0109] Example 9

[0110] Determination of the iron ion reducing capacity of vinasse extract

[0111] 1. Iron ion reducing ability

[0112] The Maotai-flavor wine lees extract of Example 1 diluted 10 times and 40 times and the Maotai-flavor wine lees extract sterilized with high temperature and high pressure were filtered through a 0.45 μm filter membrane for testing. The experimental method was carried out in accordance with the QL-SOP-YF-3-50 method.

[0113] The standard curve of ferrous sulfate detection has good linearity, y=0.0009x-0.0018, R 2 =0.9985.

[0114] The FRAP values ​​of the sauce-flavor wine lees extract of Example 1 diluted 10 times and 40 times and sterilized at high temperature and high pressure are shown in Table 6.

[0115] Table 6 FRAP values ​​of vinasse extract

[0116]

[0117] Example 10

[0118] Anti-inflammatory effects of Maotai-flavor distiller's grains extract

[0119] IL-6, or interleukin-6, is an inflammatory factor. Elevated IL-6 levels are consistent with the severity of inflammation. Testing the inhibitory effect of a test sample on IL-6 is a means of predicting its anti-inflammatory effect.

[0120] 1. Materials

[0121] Sample: the vinasse extract obtained in Example 1.

[0122] RAW264.7 (mouse monocyte-macrophage cells, BNCC342033), high-glucose DMEM medium (HyClone), PBS (HyClone), FBS (fetal bovine serum, NEWZERUM), double antibody (Gibco), IL-6 ELISA detection kit (Absin).

[0123] 2. Methods

[0124] Cell inoculation: cells were grown in complete culture medium (high glucose DMEM medium containing 10% FBS and 1% double antibody). 4 Each well of the plate was inoculated with 100 μL of culture medium and cultured in a 37°C, 5% CO2 incubator.

[0125] Sample treatment: After incubating the cells for 24 h, the culture medium was aspirated and the residual culture medium was washed with PBS. 100 μL of DMEM was added to the cells in the control and blank groups, and 100 μL of different concentrations of the vinasse extract of Example 1 (diluted with DMEM) was added to the cells in the sample group. Each group had 6 parallel cells and incubated in an incubator for 18 h.

[0126] Inflammatory stimulation: cells in the sample group and control group were stimulated with 1 μg / mL LPS, while cells in the blank group were not stimulated.

[0127] Inflammatory factor detection: After 7 hours, the culture medium was collected and the IL-6 level was detected using an ELISA kit. The IL-6 inhibition rate of the sample was calculated using the following formula:

[0128] IL-6 inhibition rate % = 100% - (c s -c b ) / (c c -c b )×100%;

[0129] c s 、c c and c b The IL-6 levels of the sample group, control group and blank group are shown in the table below. Figure 13 .

[0130] Depend on Figure 13It can be seen that the inhibitory effect of the vinasse extract of the present invention on IL-6 gradually increased at concentrations of 0.005%, 0.01%, 0.05%, 0.1%, 0.5% and 1%, among which the inhibitory effect of 1% vinasse extract on IL-6 reached 83.24%. It can be seen that the vinasse extract of the present invention has anti-inflammatory effect.

[0131] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. An application of a Maotai-flavor distiller's grains extract in anti-inflammatory electronic cigarette oil, characterized in that: The preparation method of the sauce-flavored wine lees extract comprises: enzymolyzing the sauce-flavored wine lees with a composite enzyme, filtering, centrifuging the filtrate to obtain a supernatant, extracting the supernatant with supercritical carbon dioxide, and removing impurities from the extract to obtain the sauce-flavored wine lees extract; The composite enzyme comprises composite cellulase and hemicellulase, and the mass ratio of the composite cellulase to the hemicellulase is 1:1-2; The added mass percentage of the sauce-flavor distiller's grains extract in the anti-inflammatory electronic cigarette oil is 0.1-10%.

2. The use according to claim 1, characterized in that The enzyme activity of the complex enzyme is (3-5)×10 4 U / g.

3. The use according to claim 2, characterized in that The added amount of the complex enzyme is 1% to 2% of the mass of the sauce-flavored distiller's grains.

4. The use according to claim 2 or 3, characterized in that The conditions for the enzymatic hydrolysis are: pH value of 4.5-6.5, temperature of 45-55° C., and time of 5-7 hours.

5. The use according to claim 1, characterized in that The centrifugal speed is 9000-11000 r / min, and the time is 15-25 min.

6. The use according to claim 1, characterized in that The extraction parameters are: the pressure of the extraction kettle is 25-35 MPa, the temperature is 45-50°C, and the frequency of the carbon dioxide pump is 20-25 r / min; the pressure of the separation kettle I is 5-10 MPa, the temperature is 45-50°C, and the charging coefficient is 0.3-0.

7.

7. The use according to claim 1, characterized in that The impurity removal comprises dissolving the extract with a 90% to 98% ethanol solution by volume, centrifuging and distilling to obtain the sauce-flavored wine lees extract.

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