A multi-target co-production extraction method for garlic

Abstract

The present invention provides a multi-target co-production extraction method for garlic, which relates to the technical field of separation and purification. The extraction method provided by the present invention uses MEM combined with microbial agents and neutral protease to treat garlic, and then extracts garlic essential oil, garlic polysaccharide, garlic polyphenol and garlic flavone. This extraction method is simple and low in cost, and simultaneously realizes the extraction of the above four substances, and the yields of the extracts and the contents of the active substances are high.

Description

Technical Field

[0001] The present invention relates to the technical field of separation and purification, and in particular to a method for co-producing and extracting multiple targets from garlic. Background Art

[0002] Garlic has important edible and medicinal values and is rich in nutritional components. Garlic polysaccharide is the health function component with the highest content in garlic, accounting for 15%-25% of dry garlic. It has multiple effects such as proliferating intestinal probiotics, assisting digestion, enhancing immunity, preventing liver damage, antioxidation, and antiviral effects, and has broad practical application prospects in the fields of medicine, food, etc. Garlic essential oil is a volatile essential oil extracted from garlic, which has effects such as antibacterial, bactericidal, reducing blood lipid, preventing arteriosclerosis, preventing and treating cardiovascular and cerebrovascular diseases, and has been widely used in the fields of medicines, foods, health products, and cosmetics. In addition, garlic is also rich in substances such as polyphenols and flavonoids, which also have high medicinal values.

[0003] At present, the existing extraction methods only extract one or two beneficial substances from garlic, and the extraction yield is low, and it is impossible to make full use of garlic resources.

[0004] In view of this, the present invention is specifically proposed. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for extracting garlic extract, which is simple and efficient and can solve at least one of the above problems.

[0006] In the first aspect, the present invention provides a method for extracting garlic extract, and the garlic extract includes garlic essential oil, garlic polysaccharide, garlic polyphenol, and garlic flavonoid;

[0007] The extraction method includes the following steps:

[0008] a. Pulverize garlic with water, then add MEM combined bacteria agent and neutral protease for treatment, and then add an organic solvent to extract garlic essential oil, and separate to obtain an organic phase and an aqueous phase;

[0009] b. The organic phase obtained in step a is separated and purified to obtain garlic essential oil;

[0010] c. Perform water extraction on the aqueous phase obtained in step a, and then perform solid-liquid separation to obtain an extract and garlic residue;

[0011] d. Concentrate the extract obtained in step c and then perform alcohol precipitation. The precipitate is garlic polysaccharide, and then dry the alcohol phase to obtain garlic polyphenol;

[0012] e. Perform ethanol extraction on the garlic residue obtained in step c, and then dry the extract after solid-liquid separation to obtain garlic flavonoid;

[0013] The MEM combined microbial agent mainly consists of Rhizopus nigricans, Bacillus sporolactis, and Bacillus subtilis.

[0014] As a further technical solution, in step a, garlic and water are mixed and pulped at a mass ratio of 1:3 - 1:5.

[0015] As a further technical solution, the content of Rhizopus nigricans in the MEM combined microbial agent is 1×10 8 -3×10 8 CFU / mL, the content of Bacillus sporolactis is 2×10 7 -6×10 7 CFU / mL, and the content of Bacillus subtilis is 1×10 8 -2×10 8 CFU / mL.

[0016] As a further technical solution, the addition amount of the MEM combined microbial agent is 0.4% - 0.6% of the mass of the garlic;

[0017] The addition amount of the neutral protease is 0.1% - 0.2% of the mass of the garlic.

[0018] As a further technical solution, in step a, the temperature for treating with the MEM combined microbial agent and the neutral protease is 38 - 42°C, and the treatment time is 5 - 7h.

[0019] As a further technical solution, in step a, the organic solvent includes 60 - 90 petroleum ether.

[0020] As a further technical solution, in step c, the material-liquid ratio for water extraction is 1g:3mL - 1g:5mL;

[0021] The temperature for water extraction is 75 - 85°C;

[0022] The number of times of water extraction is 1 - 3 times, and the time for each water extraction is 0.8 - 1.2h.

[0023] As a further technical solution, in step d, the mass ratio of the concentrated liquid after concentration to the garlic is 1:0.5 - 1:1.5;

[0024] During the alcohol precipitation process, the volume ratio of the concentrated liquid to ethanol is 1:3 - 1:5.

[0025] As a further technical solution, in step e, 65% - 75% ethanol is used for extraction;

[0026] The temperature for ethanol extraction is 55 - 65°C;

[0027] The time for ethanol extraction is 0.8 - 1.2h;

[0028] The material-liquid ratio for ethanol extraction is 1 g: 3 mL - 1 g: 5 mL.

[0029] As a further technical solution, the method for solid-liquid separation includes centrifugation or filtration.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] The extraction method provided by the present invention uses MEM combined with a bacterial agent and neutral protease to treat garlic, and then adopts a specific extraction process to simultaneously extract garlic essential oil, garlic polysaccharide, garlic polyphenol and garlic flavonoid, and the yield of the extract and the content of active substances are high. This extraction method is simple, efficient and low in cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0033] Figure 1 It is the extraction flow chart of Example 1 of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0034] The following will describe the implementation schemes of the present invention in detail in combination with the embodiments and examples. However, those skilled in the art will understand that the following embodiments and examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention. Those not specified in specific conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. Those reagents or instruments not specified in the manufacturer are all conventional products that can be obtained through commercial purchase.

[0035] In a first aspect, the present invention provides a method for extracting garlic extract, and the garlic extract includes garlic essential oil, garlic polysaccharide, garlic polyphenol and garlic flavonoid;

[0036] The extraction method includes the following steps:

[0037] a. Pulverize garlic with water, then add MEM combined with a bacterial agent and neutral protease for treatment, and then add an organic solvent to extract garlic essential oil, and separate to obtain an organic phase and an aqueous phase;

[0038] b. The organic phase obtained in step a is separated and purified to obtain garlic essential oil;

[0039] The aqueous phase obtained in step c.a is subjected to water extraction, and then the extract and garlic residues are obtained through solid-liquid separation.

[0040] The extract obtained in step c is concentrated and then subjected to alcohol precipitation. The precipitate is garlic polysaccharide, and then the alcohol phase is dried to obtain garlic polyphenols.

[0041] The garlic residues obtained in step c are subjected to ethanol extraction, and then the extract is dried after solid-liquid separation to obtain garlic flavonoids.

[0042] The MEM combined microbial agent mainly consists of Rhizopus nigricans, Bacillus sporolactis, and Bacillus subtilis.

[0043] Through the research of the inventor, it is found that treating garlic with the MEM combined microbial agent and neutral protease together can further improve the extraction effects of garlic essential oil, garlic polysaccharide, garlic polyphenols, and garlic flavonoids.

[0044] The extraction method provided by the present invention can simultaneously achieve the extraction of garlic essential oil, garlic polysaccharide, garlic polyphenols, and garlic flavonoids, and the purity and yield of the extracts are high. This extraction method is simple, efficient, and low-cost.

[0045] In some alternative embodiments, in step a, garlic and water are mixed at a mass ratio of 1:3 - 1:5 and pulped.

[0046] Preferably, garlic and water are mixed at a mass ratio of 1:4 and pulped.

[0047] In some alternative embodiments, the content of Rhizopus nigricans in the MEM combined microbial agent is 1×10 8 -3×10 8 CFU / mL, the content of Bacillus sporolactis is 2×10 7 -6×10 7 CFU / mL, and the content of Bacillus subtilis is 1×10 8 -2×10 8 CFU / mL.

[0048] In some alternative embodiments, the addition amount of the MEM combined microbial agent is 0.4% - 0.6% of the mass of the garlic;

[0049] The addition amount of the neutral protease is 0.1% - 0.2% of the mass of the garlic.

[0050] In some alternative embodiments, in step a, the temperature for treating with the MEM combined microbial agent and neutral protease can be, for example, but not limited to, 38°C, 39°C, 40°C, 41°C, or 42°C, preferably 40°C, and the treatment time can be, for example, but not limited to, 5 h, 6 h, or 7 h, preferably 6 h.

[0051] By further optimizing and adjusting the condition parameters of the MEM combined bacterial agent and neutral protease, the treatment of garlic can be better achieved, which helps to improve the yield and purity of the extract.

[0052] In some alternative embodiments, in step a, the organic solvent includes 60 - 90 petroleum ether.

[0053] In some alternative embodiments, in step c, the material - liquid ratio of the water extraction can be, for example, but not limited to, 1g:3mL, 1g:4mL or 1g:5mL, and preferably 1g:4mL;

[0054] The temperature of the water extraction can be, for example, but not limited to, 75, 77°C, 79°C, 81°C, 83°C or 85°C, and preferably 80°C;

[0055] The number of times of water extraction is 1 - 3 times, and 3 times is the best. The time for each water extraction can be, for example, but not limited to, 0.8h, 0.9h, 1h, 1.1h or 1.2h, and preferably 1h.

[0056] By further optimizing and adjusting the water extraction parameters, the extraction rates of garlic polysaccharide and garlic polyphenol are improved.

[0057] In some alternative embodiments, in step d, the mass ratio of the concentrated liquid after concentration to garlic can be, for example, but not limited to, 1:0.5, 1:1 or 1:1.5, and preferably 1:1;

[0058] During the alcohol precipitation process, the volume ratio of the concentrated liquid to ethanol can be, for example, but not limited to, 1:3, 1:4 or 1:5.

[0059] By further optimizing and adjusting the alcohol precipitation parameters, the yield and purity of garlic polysaccharide are improved, and the cost is saved at the same time.

[0060] In some alternative embodiments, in step e, 65% - 75% ethanol is used for extraction, and preferably 70% ethanol is used for extraction;

[0061] The temperature of the ethanol extraction can be, for example, but not limited to, 55°C, 57°C, 59°C, 61°C, 63°C or 65°C, and preferably 60°C;

[0062] The time of the ethanol extraction can be, for example, but not limited to, 0.8h, 0.9h, 1h, 1.1h or 1.2h, and preferably 1h;

[0063] The material - liquid ratio of the ethanol extraction can be, for example, but not limited to, 1g:3mL, 1g:4mL or 1g:5mL, and preferably 1g:4mL.

[0064] By further optimizing and adjusting the parameters of alcohol extraction, the yield and purity of garlic flavonoids are improved, and the cost is saved at the same time.

[0065] In some alternative embodiments, the method for solid-liquid separation includes, but is not limited to, centrifugation or filtration, or other solid-liquid separation methods well known to those skilled in the art.

[0066] The present invention will be further illustrated below by specific examples and comparative examples. However, it should be understood that these examples are only for more detailed illustration and should not be construed as limiting the present invention in any form.

[0067] The MEM combined bacterium agent used in the following examples and comparative examples was preliminarily detected, and the content of Rhizopus nigricans in the bacterium agent was 2×10 8 CFU / mL, the content of Bacillus sporolactis was 4×10 7 CFU / mL, and the content of Bacillus subtilis was 1.6×10 8 CFU / mL.

[0068] Example 1

[0069] The extraction method of garlic extract, the extraction process is as Figure 1 shown, including the following steps:

[0070] a. Mix 1000 g of garlic and 4000 g of water to make a slurry, then add the MEM combined bacterium agent (the addition amount is 0.5% of the garlic quality) and neutral protease (the addition amount is 0.15% of the garlic quality), and treat at 40 °C for 6 h. Then add 60-90 petroleum ether for extraction, and separate to obtain an organic phase and an aqueous phase;

[0071] b. Filter and concentrate the organic phase obtained in step a (heating concentration, condition parameters: negative pressure 0.1 mp, 20-40 °C) to obtain garlic essential oil;

[0072] c. Extract the aqueous phase obtained in step a with water at 80 °C (material-liquid ratio 1 g:4 mL) 3 times, each time for 1 h; then filter to obtain an extract and garlic residue;

[0073] d. Centrifuge and concentrate the extract obtained in step c to 1000 mL, add 4000 mL of absolute ethanol for ethanol precipitation, and filter to obtain garlic polysaccharide precipitate; then spin-dry the ethanol phase to obtain garlic polyphenol extract. After detection, its main component is polyphenol;

[0074] e. Extract the garlic residue obtained in step c with 4000 mL of 70% ethanol at 60 °C for 1 h, and the extraction times are 2 times. After filtration, spin-dry the filtrate to obtain garlic flavonoid extract.

[0075] Example 2

[0076] Extraction method of garlic extract, and the extraction process is as Figure 1 shown, including the following steps:

[0077] a. Mix 1000 g of garlic and 3000 g of water to make a pulp, then add MEM combined bacteria agent (the addition amount is 0.4% of the mass of garlic) and neutral protease (the addition amount is 0.2% of the mass of garlic), treat at 38 °C for 7 h, and then add an organic solvent for extraction to separate and obtain an organic phase and an aqueous phase;

[0078] b. Filter and concentrate the organic phase obtained in step a to obtain garlic essential oil;

[0079] c. Extract the aqueous phase obtained in step a by water extraction (the material-liquid ratio is 1 g: 3 mL) 3 times at 75 °C, each time for 0.8 h; then filter to obtain an extract and garlic residues;

[0080] d. Centrifuge and concentrate the extract obtained in step c to 800 mL, add 3000 mL of absolute ethanol for alcohol precipitation, and filter to obtain garlic polysaccharide precipitate; then spin-dry the ethanol phase to obtain garlic polyphenol extract. After detection, its main component is polyphenol;

[0081] e. Extract the garlic residues obtained in step c with 3000 mL of 65% ethanol at 55 °C for 0.8 h, and the extraction times are 2 times. After filtration, spin-dry the filtrate to obtain garlic flavonoid extract.

[0082] Example 3

[0083] Extraction method of garlic extract, and the extraction process is as Figure 1 shown, including the following steps:

[0084] a. Mix 1000 g of garlic and 5000 g of water to make a pulp, then add MEM combined bacteria agent (the addition amount is 0.6% of the mass of garlic) and neutral protease (the addition amount is 0.1% of the mass of garlic), treat at 42 °C for 5 h, and then add an organic solvent for extraction to separate and obtain an organic phase and an aqueous phase;

[0085] b. Filter and concentrate the organic phase obtained in step a to obtain garlic essential oil;

[0086] c. Extract the aqueous phase obtained in step a by water extraction (the material-liquid ratio is 1 g: 5 mL) 3 times at 85 °C, each time for 1.2 h; then filter to obtain an extract and garlic residues;

[0087] d. Concentrate the extract obtained in step c by centrifugation to 1200 mL, add 5000 mL of absolute ethanol for alcohol precipitation, and filter to obtain garlic polysaccharide precipitate; then rotary evaporate the ethanol phase to obtain garlic polyphenol extract. After detection, its main component is polyphenol;

[0088] e. Extract the garlic residue obtained in step c with 5000 mL of 75% ethanol at 65 °C for 1.2 h, and the extraction is carried out 2 times. After filtration, rotary evaporate the filtrate to obtain garlic flavonoid extract.

[0089] Comparative Example 1

[0090] The difference from Example 1 is that no MEM combined bacterium agent and neutral protease are added.

[0091] Comparative Example 2

[0092] The difference from Example 1 is that in step a, 0.25% of MEM combined bacterium agent is added and no neutral protease is added.

[0093] Comparative Example 3

[0094] The difference from Example 1 is that in step a, 0.5% of MEM combined bacterium agent is added and no neutral protease is added.

[0095] Comparative Example 4

[0096] The difference from Example 1 is that in step a, 0.75% of MEM combined bacterium agent is added and no neutral protease is added.

[0097] Comparative Example 5

[0098] The difference from Example 1 is that in step a, no MEM combined bacterium agent is added, and neutral protease is replaced with 0.15% of trypsin.

[0099] Comparative Example 6

[0100] The difference from Example 1 is that in step a, no MEM combined bacterium agent is added, and neutral protease is replaced with 0.15% of flavor protease.

[0101] Comparative Example 7

[0102] The difference from Example 1 is that in step a, no MEM combined bacterium agent is added, and only 0.15% of neutral protease is added.

[0103] Comparative Example 8

[0104] The difference from Example 1 is that in step a, no MEM combined bacterium agent is added, and neutral protease is replaced with 0.15% of trypsin, 0.15% of flavor protease and 0.15% of neutral protease.

[0105] Comparative Example 9

[0106] The difference from Example 1 is that in step a, 0.5% of MEM combined bacteria agent is added, and neutral protease is replaced with 0.15% of trypsin, 0.15% of flavor protease and 0.15% of neutral protease.

[0107] Test Example 1

[0108] The methods of Example 1 and Comparative Examples 1-9 were used to extract garlic. Each extraction was repeated three times. The yield of the extract and the content of active substances were detected by a spectrophotometer, and the average value of the detection results was taken, as shown in Tables 1-3.

[0109] Table 1

[0110]

[0111] Table 2

[0112]

[0113] Table 3

[0114]

[0115] Combined with the results of Tables 1-3 above, it can be seen that compared with the treatment of garlic with MEM combined bacteria agent, trypsin, flavor protease or neutral protease alone, the combination of MEM combined bacteria agent and neutral protease to treat garlic in the present invention can further improve the yields of garlic essential oil, garlic polysaccharide, garlic polyphenol and garlic flavonoid and the content rate of active substances.

[0116] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Extraction method of garlic extract Characterized in that The garlic extract includes garlic essential oil, garlic polysaccharide, garlic polyphenol and garlic flavonoid; The extraction method includes the following steps: a. Mix garlic and water in a mass ratio of 1:3 - 1:5 to make a slurry, then add MEM combined bacteria agent and neutral protease for treatment, and then add an organic solvent to extract garlic essential oil, and separate to obtain an organic phase and an aqueous phase; The treatment temperature of the MEM combined bacteria agent and neutral protease is 38 - 42 °C, and the treatment time is 5 - 7 h; The organic solvent is 60 - 90 petroleum ether; b. The organic phase obtained in step a is separated and purified to obtain garlic essential oil; c. The aqueous phase obtained in step a is subjected to water extraction, and then solid-liquid separation is carried out to obtain an extract and garlic residue; d. The extract obtained in step c is concentrated and then subjected to alcohol precipitation, and the precipitate is garlic polysaccharide, and then the alcohol phase is dried to obtain garlic polyphenol; e. The garlic residue obtained in step c is extracted with ethanol, and then the extract is dried after solid-liquid separation to obtain garlic flavonoid; The MEM combined bacteria agent is composed of Rhizopus nigricans, Bacillus sporolactis and Bacillus subtilis; The content of Rhizopus nigricans in the MEM combined microbial agent is 1×10 8 -3×10 8 CFU / mL, the content of Bacillus sporolactis is 2×10 7 -6×10 7 CFU / mL, and the content of Bacillus subtilis is 1×10 8 -2×10 8 CFU / mL; The addition amount of the MEM combined bacteria agent is 0.4% - 0.6% of the mass of the garlic; The addition amount of the neutral protease is 0.1% - 0.2% of the mass of the garlic.

2. The extraction method according to claim 1 Characterized in that In step c, the material-liquid ratio of the water extraction is 1 g: 3 mL - 1 g: 5 mL; The temperature of the water extraction is 75 - 85 °C; The number of times of the water extraction is 1 - 3 times, and the time of each water extraction is 0.8 - 1.2 h.

3. The extraction method according to claim 1 Characterized in that In step d, the mass ratio of the concentrated liquid after concentration to the garlic is 1:0.5 - 1:1.5; During the alcohol precipitation process, the volume ratio of the concentrated liquid to ethanol is 1:3 - 1:

5.

4. The extraction method according to claim 1 Characterized in that In step e, 65% - 75% ethanol is used for extraction; The temperature of the ethanol extraction is 55 - 65 °C; The time of the ethanol extraction is 0.8 - 1.2 h; The material-liquid ratio of the ethanol extraction is 1 g: 3 mL - 1 g: 5 mL.

5. The extraction method according to claim 1 Characterized in that The method of solid-liquid separation is centrifugation or filtration.

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