Maca extract for improving energy, enhancing fertility and sobering up and preparation method thereof

Abstract

The present invention proposes a maca extract for improving energy, enhancing fertility and sobering up and a preparation method thereof, and belongs to the field of medical technology. Maca powder is subjected to enzyme-assisted treatment, the treated product is subjected to low temperature and high pressure extraction, fine filtration, unfiltered material is collected, freeze-dried, and selenium-zinc-rich maca / zymosan protein complex is obtained, and the filtrate is separated by immobilized ionic liquid to obtain maca extract 1 and maca extract 2, respectively, and through different combinations, maca extract is obtained. The present invention adopts a simple and convenient method to prepare maca extract, and different maca extracts for improving energy, enhancing fertility or sobering up are obtained by different combinations, respectively, with high content of active substances, and can be combined according to different needs, with broad application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to a maca extract capable of improving energy, enhancing fertility and sobering up, and a preparation method thereof. Background Art

[0002] Maca (Lepdium meyenii Walpers, Maca) is an annual or biennial herbaceous plant of the genus Lepidium in the family Brassicaceae, native to the Andes of Peru. Maca is not only rich in protein, unique amino acid composition, multiple unsaturated fatty acids, mineral elements and vitamins, but also contains secondary metabolites such as aromatic glucosinolates, macaamides, macaene, sterols, adenosine, saponins, imidazole alkaloids, etc. It has a variety of health functions and therapeutic effects, such as enhancing energy, resisting fatigue, improving fertility, relieving stress, improving sexual function, improving sleep, neuroprotection, enhancing immunity and anti-tumor.

[0003] Traditional maca extract preparation methods include water extraction, alcohol extraction, alcohol-water extraction, water extraction and alcohol precipitation, and alcohol extraction and water precipitation. It is difficult to fully obtain the anti-fatigue active ingredients of maca. Given the complexity of maca's biologically active ingredients, existing maca extracts have the disadvantages of low content of active ingredients, low potency, and high cost. Therefore, there is an urgent need for an extraction method for maca extracts with high content of active ingredients, high utilization rate, and high activity of the obtained extract. Summary of the invention

[0004] The purpose of the present invention is to provide a maca extract for improving energy, enhancing fertility and sobering up, and a preparation method thereof. The maca extract is prepared by a simple and convenient method, and different maca extracts for improving energy, enhancing fertility or sobering up are obtained through different combinations. The extracts have a high content of active substances and can be combined according to different needs, and have broad application prospects.

[0005] The technical solution of the present invention is achieved in this way:

[0006] The present invention provides a method for preparing a maca extract capable of improving energy, enhancing fertility and sobering up, comprising the following steps:

[0007] S1. Add maca powder to water, add complex enzyme, stir, inoculate selenium-rich yeast and zinc-rich yeast seed liquid for pre-treatment, and extract the treated liquid at low temperature and high pressure to obtain an extract;

[0008] S2. The extract was finely filtered through a ceramic membrane, the unfiltered material was collected, freeze-dried, and a selenium-rich-zinc maca / zymosan protein complex was obtained, and the filtrate was retained;

[0009] S3. Add the immobilized ionic liquid to the filtrate, stir and adsorb under the action of an external magnetic field, separate with a magnet, collect the liquid and the solid, wherein the liquid is freeze-dried to obtain maca extract 1; the solid is added to ethanol and heated for desorption, the solid is separated with a magnet, washed, dried, and recycled, the liquid is recovered as ethanol, and freeze-dried to obtain maca extract 2;

[0010] S4. The maca extract 1 and the selenium-zinc-rich maca / yeast polysaccharide protein complex are mixed evenly to prepare a maca extract that enhances energy and fertility;

[0011] Or, using maca extract 2 as a hangover-relieving maca extract;

[0012] Or the maca extract 1, the maca extract 2 and the selenium-zinc-rich maca / yeast polysaccharide protein complex are uniformly mixed to obtain a maca extract that enhances energy, strengthens fertility and relieves alcohol.

[0013] As a further improvement of the present invention, the mass ratio of maca powder and complex enzyme in step S1 is 10:1-2, the complex enzyme is selected from at least two of cellulase, pectinase, amylase, neutral protease, trypsin, alkaline protease, ficin, papain and bromelain, the temperature of the enzymolysis is 35-45°C, the time is 0.5-1.5h, the inoculation amount of the selenium-enriched yeast and zinc-enriched yeast seed liquid is 1-3v / v% and 1.5-2.5v / v%, respectively, the pretreatment conditions are 25-32°C, 150-250r / min, enzyme-assisted fermentation culture is 36-48h, the temperature of the low-temperature and high-pressure extraction is 5-25°C, the pressure is 20-50MPa, and the extraction time is 1-2h.

[0014] As a further improvement of the present invention, the composite enzyme is cellulase and neutral protease, and the mass ratio is 10:3-5.

[0015] As a further improvement of the present invention, the preparation method of the selenium-enriched yeast and zinc-enriched yeast seed solution is as follows: the selenium-enriched yeast and zinc-enriched yeast are inoculated into Gao's medium respectively, and the activation culture is carried out at 28-32°C, 100-200r / min, and the activation culture is carried out for 18-24h to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of selenium-enriched yeast and zinc-enriched yeast seed liquid.

[0016] As a further improvement of the present invention, the pore size of the ceramic membrane in step S2 is 200-500 nm.

[0017] As a further improvement of the present invention, the preparation method of the immobilized ionic liquid in step S3 is as follows:

[0018] T1. Preparation of magnetic particles: Under the protection of inert gas, ferric chloride and ferrous chloride are dissolved in water, ammonia water is added dropwise, stirred and heated for reaction, centrifuged, washed, dried, and calcined to obtain magnetic particles;

[0019] T2. Preparation of modified magnetic particles: adding magnetic particles to water, adding dopamine hydrochloride and a catalyst, heating and stirring the reaction, separating with a magnet, washing, and drying to obtain modified magnetic particles;

[0020] T3. N-methylimidazole and 3-chloropropyltrimethoxysilane, silane coupling agent KH560 were mixed, heated and stirred to react, water was added for hydrolysis, filtered, washed, and dried to obtain fixed ionic liquid particles;

[0021] T4. Add the immobilized ionic liquid particles, modified magnetic particles and salt into acetonitrile, stir to react, filter, wash and dry to obtain the immobilized ionic liquid.

[0022] As a further improvement of the present invention, the mass ratio of ferric chloride, ferrous chloride and ammonia water in step T1 is 3.24:1.26:5-10, the temperature of the stirring and heating reaction is 80-90°C, the time is 3-5h, the temperature of the calcination is 400-500°C, and the time is 1-3h; the mass ratio of the magnetic particles, dopamine hydrochloride and catalyst in step T2 is 10:2-3:0.1-0.3, the pH of the catalyst is Tris-HCl solution of 8.5-9.5, and the heating and stirring reaction is The reaction temperature is 35-45°C and the reaction time is 3-5h; the molar ratio of N-methylimidazole, 3-chloropropyltrimethoxysilane and silane coupling agent KH560 in step T3 is 1:1:0.2-0.3, the temperature of the heating and stirring reaction is 100-110°C and the reaction time is 20-24h; the mass ratio of the fixed ionic liquid particles, modified magnetic particles and salt in step T4 is 5-7:10:7-12, the stirring reaction time is 22-24h, and the salt is NaBF4 and KPF6.

[0023] As a further improvement of the present invention, the solid-liquid ratio of the immobilized ionic liquid and the filtrate in step S3 is 1:5-10 g / mL, the strength of the external magnetic field is 0.03-0.05 T, the temperature of the heating desorption is 40-50° C., and the time is 30-50 min.

[0024] As a further improvement of the present invention, the mass ratio of maca extract 1 and selenium-zinc-rich maca / zymosan protein complex in the maca extract for improving energy and fertility in step S4 is 3-5:10, and the mass ratio of maca extract 1, maca extract 2 and selenium-zinc-rich maca / zymosan protein complex in the maca extract for improving energy, enhancing fertility and sobering up is 3-5:2-4:10.

[0025] The present invention further protects a maca extract for improving energy and fertility, or a maca extract for sobering up, or a maca extract for improving energy, fertility and sobering up, obtained by the above-mentioned preparation method.

[0026] The present invention has the following beneficial effects:

[0027] The present invention adopts a method of compound enzyme + zinc-rich yeast / selenium-rich yeast + low temperature and high pressure to extract maca extract, the compound enzyme is cellulase and neutral protease, can greatly destroy the maca plant cell wall, release active substances, neutral protease enzymolysis maca protein, obtain high-activity protein, further under the action of selenium-rich yeast and zinc-rich yeast, maca polysaccharide and maca protein are converted into selenium-rich / zinc polysaccharide and selenium-rich / zinc protein, selenium is a powerful antioxidant, for the immune system and thyroid function are very important, lack of selenium may cause fatigue. Zinc is essential for the immune system, cell division and protein synthesis, lack of zinc may cause fatigue and decreased immunity. The present invention, on the basis of maca polysaccharide and maca protein, makes the surface of polysaccharide and protein molecules enriched with organic zinc and organic selenium, greatly improves the product anti-fatigue, improves energy, improves immunity and improves fertility. At the same time, under the action of low temperature and high pressure extraction, it can further promote the breaking of plant cell walls and yeast cell walls, so that a large amount of active components in yeast can be dissolved, thereby obtaining an extract with high protein and high polysaccharide content.

[0028] The extract is filtered through a ceramic membrane with a pore size of 200-500nm, and the permeates are mainly large molecular polysaccharides and proteins. They are freeze-dried to produce a selenium-zinc-rich maca / yeast polysaccharide protein complex, which has good effects in resisting fatigue, improving energy, enhancing immunity and enhancing fertility.

[0029] The filtrate contains rich active components such as maca flavonoids, polyphenols, triterpenes, saponins, etc. Under the action of immobilized ionic liquid and the support of an external magnetic field, these active components move and are fixedly adsorbed on the immobilized ionic liquid particles. After solid-liquid separation, they are added to the good solvent ethanol and heated to promote desorption, so that these active components enter the ethanol. The immobilized ionic liquid solid is separated, washed and dried, and can be reused. The extract is recovered by ethanol and dried to obtain maca extract 2, which has a good alcohol-removing effect.

[0030] In addition, the unseparated liquid is rich in amino acids, small molecule sugars and other substances. After drying, maca extract 1 is obtained, which is mixed with selenium-zinc-rich maca / yeast polysaccharide protein complex to obtain maca extract that enhances energy and fertility.

[0031] The prepared maca extract 1, maca extract 2, and selenium-zinc-rich maca / yeast polysaccharide protein complex are mixed to prepare a maca extract capable of improving energy, enhancing fertility, and sobering up.

[0032] The immobilized ionic liquid prepared by the invention is prepared by modifying the surface of the magnetic ferroferric oxide particles by polydopamine, reacting and coupling N-methylimidazole and 3-chloropropyltrimethoxysilane, and then coupling with a silane coupling agent KH560. During the hydrolysis process, a layer of silicon dioxide shell is formed, thereby preparing silicon dioxide particles with epoxy groups and N-methylimidazole, which can react with magnetic particles modified by polydopamine with amino groups. After being exchanged with salts NaBF4 and KPF6, the prepared immobilized ionic liquid has both the characteristics of a solid carrier and the unique advantages of an ionic liquid, can be used for the adsorption and separation of active ingredients, and does not have the problem of residual ionic liquid.

[0033] The invention adopts a simple and convenient method to prepare maca extract, and obtains different maca extracts for improving energy, enhancing fertility or sobering up through different combinations. The extracts have high content of active substances, can be combined according to different needs, and have broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0035] Figure 1 This is a 800-fold micrograph of the maca powder in Example 1 of the present invention;

[0036] Figure 2 This is a 800-fold micrograph of the extract in Example 1 of the present invention. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] Selenium-enriched yeast, 20 billion cfu / g, was purchased from Angel Yeast Co., Ltd. Zinc-enriched yeast, 20 billion cfu / g, was purchased from Fufeng Snort Biotechnology Co., Ltd.

[0039] The preparation method of selenium-enriched yeast and zinc-enriched yeast seed liquid is as follows: Selenium-enriched yeast and zinc-enriched yeast are inoculated into Gao's medium respectively, and activated and cultured for 18-24 hours at 30°C and 150r / min to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of selenium-enriched yeast and zinc-enriched yeast seed liquid.

[0040] Preparation Example 1 Preparation of immobilized ionic liquid

[0041] Here’s how:

[0042] T1. Preparation of magnetic particles: Under nitrogen protection, 3.24 g of ferric chloride and 1.26 g of ferrous chloride were dissolved in 200 mL of water, 5 g of ammonia water was added dropwise, heated to 80 ° C, stirred for 3 h, centrifuged, washed, dried, and calcined at 400 ° C for 1 h to obtain magnetic particles;

[0043] T2. Preparation of modified magnetic particles: 10 g of magnetic particles were added to 200 mL of water, 2 g of dopamine hydrochloride and 0.1 g of catalyst were added, heated to 35°C, stirred for reaction for 3 h, separated by magnet, washed, and dried to obtain modified magnetic particles;

[0044] The catalyst has a pH of 8.5 in a Tris-HCl solution;

[0045] T3. Mix 10mmol N-methylimidazole, 10mmol 3-chloropropyltrimethoxysilane and 2mmol silane coupling agent KH560, heat to 100°C, stir and react for 20h, cool to room temperature, add 100mL water, hydrolyze for 1h, filter, wash and dry to obtain fixed ionic liquid particles;

[0046] T4. Add 5 g of immobilized ionic liquid particles, 10 g of modified magnetic particles and 7 g of NaBF4 into 200 mL of acetonitrile, stir and react for 22 h, filter, wash and dry to obtain an immobilized ionic liquid.

[0047] Preparation Example 2 Preparation of immobilized ionic liquid

[0048] Here’s how:

[0049] T1. Preparation of magnetic particles: Under nitrogen protection, 3.24 g of ferric chloride and 1.26 g of ferrous chloride were dissolved in 200 mL of water, 10 g of ammonia water was added dropwise, heated to 90 ° C, stirred for 5 h, centrifuged, washed, dried, and calcined at 500 ° C for 3 h to obtain magnetic particles;

[0050] T2. Preparation of modified magnetic particles: 10 g of magnetic particles were added to 200 mL of water, 3 g of dopamine hydrochloride and 0.3 g of catalyst were added, heated to 45°C, stirred for reaction for 5 h, separated by magnet, washed, and dried to obtain modified magnetic particles;

[0051] The catalyst has a pH of 9.5 in a Tris-HCl solution;

[0052] T3. Mix 10mmol N-methylimidazole, 10mmol 3-chloropropyltrimethoxysilane and 3mmol silane coupling agent KH560, heat to 110°C, stir and react for 24h, cool to room temperature, add 100mL water, hydrolyze for 1h, filter, wash and dry to obtain fixed ionic liquid particles;

[0053] T4. Add 7 g of fixed ionic liquid particles, 10 g of modified magnetic particles and 12 g of NaBF4 into 200 mL of acetonitrile, stir and react for 24 h, filter, wash and dry to obtain an immobilized ionic liquid.

[0054] Preparation Example 3 Preparation of immobilized ionic liquid

[0055] Here’s how:

[0056] T1. Preparation of magnetic particles: Under nitrogen protection, 3.24 g of ferric chloride and 1.26 g of ferrous chloride were dissolved in 200 mL of water, 7 g of ammonia water was added dropwise, heated to 85 ° C, stirred for 4 h, centrifuged, washed, dried, and calcined at 450 ° C for 2 h to obtain magnetic particles;

[0057] T2. Preparation of modified magnetic particles: 10 g of magnetic particles were added to 200 mL of water, 2.5 g of dopamine hydrochloride and 0.2 g of catalyst were added, heated to 40°C, stirred for reaction for 4 h, separated by magnet, washed, and dried to obtain modified magnetic particles;

[0058] A Tris-HCl solution of the catalyst with a pH of 9;

[0059] T3. Mix 10mmol N-methylimidazole, 10mmol 3-chloropropyltrimethoxysilane and 2.5mmol silane coupling agent KH560, heat to 105°C, stir and react for 22h, cool to room temperature, add 100mL water, hydrolyze for 1h, filter, wash and dry to obtain fixed ionic liquid particles;

[0060] T4. Add 6 g of immobilized ionic liquid particles, 10 g of modified magnetic particles and 10 g of NaBF4 into 200 mL of acetonitrile, stir and react for 23 h, filter, wash and dry to obtain an immobilized ionic liquid.

[0061] Comparative Preparation Example 1

[0062] Compared with Preparation Example 3, the difference is that step T2 is not performed.

[0063] The details are as follows:

[0064] T1. Preparation of magnetic particles: Under nitrogen protection, 3.24 g of ferric chloride and 1.26 g of ferrous chloride were dissolved in 200 mL of water, 7 g of ammonia water was added dropwise, heated to 85 ° C, stirred for 4 h, centrifuged, washed, dried, and calcined at 450 ° C for 2 h to obtain magnetic particles;

[0065] T2. Mix 10mmol N-methylimidazole, 10mmol 3-chloropropyltrimethoxysilane, and 2.5mmol silane coupling agent KH560, heat to 105°C, stir and react for 22h, cool to room temperature, add 100mL water, hydrolyze for 1h, filter, wash, and dry to obtain fixed ionic liquid particles;

[0066] T3. Add 6 g of immobilized ionic liquid particles, 10 g of magnetic particles and 10 g of NaBF4 into 200 mL of acetonitrile, stir and react for 23 h, filter, wash and dry to obtain an immobilized ionic liquid.

[0067] Comparative Preparation Example 2

[0068] Compared with Preparation Example 3, the difference is that the silane coupling agent KH560 is not added in step T3.

[0069] The details are as follows:

[0070] T1. Preparation of magnetic particles: Under nitrogen protection, 3.24 g of ferric chloride and 1.26 g of ferrous chloride were dissolved in 200 mL of water, 7 g of ammonia water was added dropwise, heated to 85 ° C, stirred for 4 h, centrifuged, washed, dried, and calcined at 450 ° C for 2 h to obtain magnetic particles;

[0071] T2. Preparation of modified magnetic particles: 10 g of magnetic particles were added to 200 mL of water, 2.5 g of dopamine hydrochloride and 0.2 g of catalyst were added, heated to 40°C, stirred for reaction for 4 h, separated by magnet, washed, and dried to obtain modified magnetic particles;

[0072] A Tris-HCl solution of the catalyst with a pH of 9;

[0073] T3. Mix 10 mmol N-methylimidazole and 12.5 mmol 3-chloropropyltrimethoxysilane, heat to 105°C, stir and react for 22 hours, cool to room temperature, add 100 mL water, hydrolyze for 1 hour, filter, wash, and dry to obtain fixed ionic liquid particles;

[0074] T4. Add 6 g of immobilized ionic liquid particles, 10 g of modified magnetic particles and 10 g of NaBF4 into 200 mL of acetonitrile, stir and react for 23 h, filter, wash and dry to obtain an immobilized ionic liquid.

[0075] Example 1

[0076] This embodiment provides a method for preparing a maca extract for improving energy, enhancing fertility, and sobering up, comprising the following steps:

[0077] S1. Mix 10g maca powder (such as Figure 1 ) was added to 500 mL of water, 1 g of complex enzyme was added, and enzymolysis was carried out at 35 ° C for 0.5 h. The seed liquid of selenium-enriched yeast and zinc-enriched yeast was inoculated at an inoculation rate of 1 v / v% and 1.5 v / v%, respectively. The enzyme-assisted fermentation culture was carried out at 25 ° C and 150 r / min for 36 h. The fermentation product was subjected to a temperature of 5 ° C and a pressure of 50 MPa for 1 h to obtain an extract (such as Figure 2 ); Figure 1 The microscopic image of maca powder shows that there are a large number of complete cell structures in maca powder, while the prepared extract has almost no cell structure. The cells are completely broken, the nutrients in the cells are completely released, and the effective ingredients are fully extracted.

[0078] The complex enzyme is cellulase and neutral protease, with a mass ratio of 10:3;

[0079] S2. The extract was finely filtered through a ceramic membrane with a pore size of 200 nm, the unfiltered material was collected, freeze-dried, and a selenium-rich-zinc maca / zymosan protein complex was obtained, and the filtrate was retained;

[0080] S3. Add 10 g of the immobilized ionic liquid prepared in Preparation Example 1 to 50 mL of the filtrate, stir and adsorb for 1 h under the action of an external magnetic field with a strength of 0.03 T, separate with a magnet, collect the liquid and the solid, wherein the liquid is freeze-dried to obtain maca extract 1; add the solid to 200 mL of ethanol and heat to 40° C., desorb for 30 min, separate the solid with a magnet, wash, dry, and recycle, recover ethanol from the liquid, and freeze-dry to obtain maca extract 2;

[0081] S4. 3 g of maca extract 1 and 10 g of selenium-zinc-rich maca / yeast polysaccharide protein complex were stirred and mixed for 10 min to obtain a maca extract that enhances energy and fertility;

[0082] Or, using maca extract 2 as a hangover-relieving maca extract;

[0083] Alternatively, 3 g of maca extract 1, 2 g of maca extract 2 and 10 g of selenium-zinc-rich maca / yeast polysaccharide protein complex are stirred and mixed for 10 minutes to prepare a maca extract that improves energy, enhances fertility and relieves alcohol.

[0084] Example 2

[0085] This embodiment provides a method for preparing a maca extract for improving energy, enhancing fertility, and sobering up, comprising the following steps:

[0086] S1. Add 10g maca powder to 500mL water, add 2g compound enzyme, enzymolysis at 45℃ for 1.5h, inoculate selenium-enriched yeast and zinc-enriched yeast seed liquid, the inoculation amount is 3v / v% and 2.5v / v%, respectively, at 32℃, 250r / min, enzyme-assisted fermentation culture for 48h, the fermentation product is subjected to temperature of 25℃, pressure of 20MPa, extraction for 2h, and obtain the extract;

[0087] The complex enzyme is cellulase and neutral protease, with a mass ratio of 10:5;

[0088] S2. The extract was finely filtered through a ceramic membrane with a pore size of 500 nm, the unfiltered material was collected, freeze-dried, and a selenium-rich-zinc maca / zymosan protein complex was obtained, and the filtrate was retained;

[0089] S3. Add 10 g of the immobilized ionic liquid prepared in Preparation Example 2 to 100 mL of the filtrate, stir and adsorb for 1 h under the action of an external magnetic field with a strength of 0.05 T, separate with a magnet, collect the liquid and the solid, wherein the liquid is freeze-dried to obtain maca extract 1; add the solid to 200 mL of ethanol and heat to 50° C., desorb for 50 min, separate the solid with a magnet, wash, dry, and recycle, recover ethanol from the liquid, and freeze-dry to obtain maca extract 2;

[0090] S4. 5 g of maca extract 1 and 10 g of selenium-zinc-rich maca / yeast polysaccharide protein complex were stirred and mixed for 10 min to obtain a maca extract that enhances energy and fertility;

[0091] Or, using maca extract 2 as a hangover-relieving maca extract;

[0092] Alternatively, 5 g of maca extract 1, 4 g of maca extract 2 and 10 g of selenium-zinc-rich maca / yeast polysaccharide protein complex are stirred and mixed for 10 minutes to prepare a maca extract that improves energy, enhances fertility and relieves alcohol.

[0093] Example 3

[0094] This embodiment provides a method for preparing a maca extract for improving energy, enhancing fertility, and sobering up, comprising the following steps:

[0095] S1. Add 10g maca powder to 500mL water, add 1.5g compound enzyme, enzymolysis at 40℃ for 1h, inoculate selenium-enriched yeast and zinc-enriched yeast seed liquid, the inoculation amount is 2v / v% and 2v / v%, respectively, at 29℃, 200r / min, enzyme-assisted fermentation culture for 42h, the fermentation product is subjected to temperature of 15℃, pressure of 35MPa, extraction for 1.5h, and obtain the extract;

[0096] The complex enzyme is cellulase and neutral protease, with a mass ratio of 10:4;

[0097] S2. The extract was finely filtered through a ceramic membrane with a pore size of 300 nm, the unfiltered material was collected, freeze-dried, and a selenium-rich-zinc maca / zymosan protein complex was obtained, and the filtrate was retained;

[0098] S3. Add 10 g of the immobilized ionic liquid prepared in Preparation Example 3 to 70 mL of the filtrate, stir and adsorb for 1 h under the action of an external magnetic field with a strength of 0.04 T, separate with a magnet, collect the liquid and the solid, wherein the liquid is freeze-dried to obtain maca extract 1; add the solid to 200 mL of ethanol and heat to 45° C., desorb for 40 min, separate the solid with a magnet, wash, dry, and recycle, recover ethanol from the liquid, and freeze-dry to obtain maca extract 2;

[0099] S4. 4 g of maca extract 1 and 10 g of selenium-zinc-rich maca / yeast polysaccharide protein complex were stirred and mixed for 10 min to obtain a maca extract that enhances energy and fertility;

[0100] Or, maca extract 2 is used as a maca extract for hangover relief;

[0101] Alternatively, 4 g of maca extract 1, 3 g of maca extract 2 and 10 g of selenium-zinc-rich maca / yeast polysaccharide protein complex are stirred and mixed for 10 minutes to prepare a maca extract that improves energy, enhances fertility and relieves alcohol.

[0102] Example 4

[0103] Compared with Example 3, the difference is that the composite enzyme is a single cellulase.

[0104] Example 5

[0105] Compared with Example 3, the difference is that the complex enzyme is a single neutral protease.

[0106] Comparative Example 1

[0107] Compared with Example 3, the difference is that no complex enzyme is added in step S1.

[0108] The details are as follows:

[0109] S1. Add 10g maca powder to 500mL water, inoculate with selenium-enriched yeast and zinc-enriched yeast seed liquid, the inoculation amount is 2v / v% and 2v / v%, respectively, ferment and culture at 29℃, 200r / min, ferment for 42h, the fermentation product is subjected to temperature of 15℃, pressure of 35MPa, extraction for 1.5h, and obtain the extract.

[0110] Comparative Example 2

[0111] Compared with Example 3, the difference is that no selenium-enriched yeast seed liquid is inoculated in step S1.

[0112] The details are as follows:

[0113] S1. Add 10g maca powder to 500mL water, add 1.5g compound enzyme, enzymolysis at 40℃ for 1h, inoculate zinc-rich yeast seed liquid with an inoculation amount of 4v / v%, 29℃, 200r / min, enzyme-assisted fermentation culture for 42h, the fermentation product is subjected to temperature of 15℃, pressure of 35MPa, extraction for 1.5h, and obtain the extract;

[0114] The complex enzyme is cellulase and neutral protease, and the mass ratio is 10:4.

[0115] Comparative Example 3

[0116] Compared with Example 3, the difference is that the zinc-enriched yeast seed liquid is not inoculated in step S1.

[0117] The details are as follows:

[0118] S1. Add 10g maca powder to 500mL water, add 1.5g compound enzyme, enzymolysis at 40℃ for 1h, inoculate selenium-enriched yeast seed liquid with an inoculation amount of 4v / v%, 29℃, 200r / min, enzyme-assisted fermentation culture for 42h, the fermentation product is subjected to temperature of 15℃, pressure of 35MPa, extraction for 1.5h to obtain an extract;

[0119] The complex enzyme is cellulase and neutral protease, and the mass ratio is 10:4.

[0120] Comparative Example 4

[0121] Compared with Example 3, the difference is that the selenium-enriched yeast and zinc-enriched yeast seed liquid are not inoculated in step S1.

[0122] The details are as follows:

[0123] S1. Add 10g maca powder to 500mL water, add 1.5g compound enzyme, enzymolyze at 40℃ for 1h, extract the product at 15℃ and 35MPa for 1.5h to obtain the enzymolyzate;

[0124] The complex enzyme is cellulase and neutral protease, and the mass ratio is 10:4.

[0125] Comparative Example 5

[0126] Compared with Example 3, the difference is that no external magnetic field is applied in step S3.

[0127] The details are as follows:

[0128] S3. Add 10 g of the immobilized ionic liquid prepared in Preparation Example 3 to 70 mL of the filtrate, stir and adsorb for 1 hour, separate with a magnet, collect the liquid and the solid, wherein the liquid is freeze-dried to obtain maca extract 1; add the solid to 200 mL of ethanol and heat to 45°C, desorb for 40 minutes, separate the solid with a magnet, wash, dry, and recycle, recover ethanol from the liquid, and freeze-dry to obtain maca extract 2.

[0129] Comparative Example 6

[0130] Compared with Example 3, the difference is that step S3 is not performed and the filtrate is extracted with dichloromethane.

[0131] The details are as follows:

[0132] S3. Add 200 mL of dichloromethane to 70 mL of the filtrate for extraction, separate the liquids, freeze-dry the aqueous phase to obtain maca extract 1; remove the solvent from the dichloromethane phase under reduced pressure, and dry to obtain maca extract 2.

[0133] Comparative Example 7

[0134] Compared with Example 3, the difference is that the immobilized ionic liquid is prepared by Comparative Preparation Example 1.

[0135] Comparative Example 8

[0136] Compared with Example 3, the difference is that the immobilized ionic liquid is prepared by Comparative Preparation Example 2.

[0137] Test Example 1

[0138] The maca extracts for improving energy and fertility, the maca extracts for sobering up, and the maca extracts for improving energy, fertility and sobering up prepared in Examples 1-5 and Comparative Examples 1-8 were respectively prepared into sample solutions with a welding layer of 0.02 mg / mL.

[0139] A DPPH free radical solution with a concentration of 0.1 mmol / L was prepared using 95% ethanol solution and stored at 4°C away from light.

[0140] Mix 2 mL of sample solution with 2 mL of DPPH free radical solution, shake thoroughly, and react for 30 minutes in the dark. Measure the absorbance at a wavelength of 517 nm. Under the same conditions, no DPPH was added as the background group, and no sample solution was added as the blank group. The scavenging rate of DPPH free radicals was calculated according to the following formula. The same concentration of vitamin C was used as the positive control, and the measurement was repeated 3 times. The results are shown in Table 1.

[0141] DPPH free radical scavenging rate (%) = [1-(A s -A b ) / A c ]×100%

[0142] Where: A s is the absorbance value of the sample group; A b is the absorbance value of the background group; A c is the absorbance value of the blank group.

[0143] Table 1

[0144]

[0145] It can be seen from the above table that the maca extracts for improving energy and fertility, or the maca extracts for sobering up, or the maca extracts for improving energy, fertility and sobering up prepared in Examples 1-3 of the present invention have good antioxidant effects.

[0146] Test Example 2

[0147] 140 healthy ICR female mice were selected for the experiment, and the body weight was 18-22g at the end of the adaptation period. The animals were divided into 14 experimental groups, with 10 mice in each experimental group, namely the negative control group, Example 1-5 group, and Comparative Example 1-8 group. Water was freely available. The Example 1-5 and Comparative Example 1-8 groups were given the corresponding maca extracts for improving energy and enhancing fertility, with an intragastric dose of 0.5g / (kg·BW), and the negative control group was replaced with an equal amount of deionized water. The intragastric administration was continued for 30 days. 30 minutes after the last intragastric administration, a lead wire of 5% of the body weight was tied to the base of the mouse tail, and the mouse was placed in a swimming box with a depth of about 30cm to swim. The water temperature was controlled at (25±1.0)℃, and the time from the start of swimming to the death of the mouse was recorded. The results are shown in Table 2.

[0148] Table 2

[0149]

[0150] Note: * Compared with the negative control group, P < 0.05.

[0151] As can be seen from the above table, the maca extracts for improving energy and enhancing fertility prepared in Examples 1-3 of the present invention can significantly prolong the weighted swimming time of mice.

[0152] Test Example 3

[0153] Take SPF grade C57BL / 6J mice, body weight 18-22g, male, adaptive feeding for 1 week, free drinking water and food. Randomly divided into blank group, model group, bifendate group, embodiment 1-5, comparative example 1-8 group, 10 in each group. Every day, embodiment 1-5, comparative example 1-8 group gavage to embodiment 1-5 and comparative example 1-8 prepared alcohol-relieving maca extract, gavage amount is 0.5g / kg, blank group, model group with equal amount of deionized water replacement, bifendate group to give 150mg / kg bifendate. After the last administration, except blank group, other groups of mice were given 1 gavage of 50% ethanol, gavage amount 15mL / kg, establish mouse acute alcoholic liver disease model, record mouse drunkenness, sleep, sober up time. The mouse back down for 30s is the disappearance of righting reflex; the mouse back down within 30s can be righted for righting reflex recovery. The incubation time of drunkenness is the time from oral administration to the disappearance of righting reflex; the sobering time is the time from the disappearance of righting reflex to its recovery. The results are shown in Table 3.

[0154] Table 3

[0155]

[0156] Note: * compared with the blank group, P < 0.05, # compared with the model group, P < 0.05.

[0157] It can be seen from the above table that the hangover-relieving maca extracts prepared in Examples 1-3 of the present invention can significantly prolong the latent time of drunkenness and shorten the sobering time.

[0158] After fasting for 16 hours but not drinking water, blood was collected from the abdominal aorta of the animals after anesthesia, and the activity of GPT (alanine aminotransferase) and GOT (aspartate aminotransferase) was tested according to the instructions of the kit. An appropriate amount of liver tissue was weighed and ground with 9 times normal saline, and then the grinding liquid was centrifuged to obtain the supernatant, and the levels of SOD (superoxide dismutase) and MDA (malondialdehyde) were tested according to the instructions of the kit. The results are shown in Table 4.

[0159] Table 4

[0160]

[0161] Note: * compared with the blank group, P < 0.05, # compared with the model group, P < 0.05.

[0162] It can be seen from the above table that the hangover-relieving maca extracts prepared in Examples 1-3 of the present invention can significantly reduce the activities of GPT and GOT, increase the activity of SOD, and reduce the content of MDA.

[0163] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a maca extract for improving energy and sobering up, characterized in that: The following steps are involved: S1. Add maca powder to water, add complex enzyme, stir for enzymolysis, inoculate selenium-rich yeast and zinc-rich yeast seed liquid for pre-treatment, and extract the treated liquid at low temperature and high pressure to obtain an extract; The complex enzyme is cellulase and neutral protease, with a mass ratio of 10:3-5; S2. The extract was finely filtered through a ceramic membrane, the unfiltered material was collected, freeze-dried, and a selenium-rich-zinc maca / zymosan protein complex was obtained, and the filtrate was retained; S3. Add the immobilized ionic liquid to the filtrate, stir and adsorb under the action of an external magnetic field, separate with a magnet, collect the liquid and the solid, wherein the liquid is freeze-dried to obtain maca extract 1; the solid is added to ethanol and heated for desorption, the solid is separated with a magnet, washed, dried, and recycled, the liquid is recovered as ethanol, and freeze-dried to obtain maca extract 2; The preparation method of the immobilized ionic liquid is as follows: T1. Preparation of magnetic particles: Under the protection of inert gas, ferric chloride and ferrous chloride are dissolved in water, ammonia water is added dropwise, stirred and heated for reaction, centrifuged, washed, dried, and calcined to obtain magnetic particles; T2. Preparation of modified magnetic particles: adding magnetic particles to water, adding dopamine hydrochloride and a catalyst, heating and stirring the reaction, separating with a magnet, washing, and drying to obtain modified magnetic particles; T3. N-methylimidazole and 3-chloropropyltrimethoxysilane, silane coupling agent KH560 were mixed, heated and stirred to react, water was added for hydrolysis, filtered, washed, and dried to obtain fixed ionic liquid particles; T4. The fixed ionic liquid particles, modified magnetic particles and salt were added to acetonitrile, stirred for reaction, filtered, washed and dried to obtain an immobilized ionic liquid; S4. The maca extract 1 and the selenium-zinc-rich maca / zymosan protein complex are mixed evenly to obtain a maca extract for enhancing energy; Or, using maca extract 2 as a hangover-relieving maca extract; Alternatively, the maca extract 1, the maca extract 2 and the selenium-zinc-rich maca / yeast polysaccharide protein complex are uniformly mixed to obtain a maca extract for enhancing energy and sobering up.

2. The preparation method according to claim 1, characterized in that: In step S1, the mass ratio of maca powder and complex enzyme is 10:1-2, the temperature of the enzymatic hydrolysis is 35-45°C, the time is 0.5-1.5h, the inoculation amounts of the selenium-enriched yeast and zinc-enriched yeast seed solutions are 1-3v / v% and 1.5-2.5v / v%, respectively, the pretreatment conditions are 25-32°C, 150-250r / min, enzyme-assisted fermentation culture is 36-48h, the temperature of the low-temperature and high-pressure extraction is 5-25°C, the pressure is 20-50MPa, and the extraction time is 1-2h.

3. The preparation method according to claim 1, characterized in that: The preparation method of the selenium-enriched yeast and zinc-enriched yeast seed liquid is as follows: the selenium-enriched yeast and zinc-enriched yeast are inoculated into Gao's medium respectively, and the activation culture is carried out at 28-32°C, 100-200r / min, and the activation culture is carried out for 18-24h to obtain a culture medium with a bacterial content of 10 8 -10 9 cfu / mL of selenium-enriched yeast and zinc-enriched yeast seed liquid.

4. The preparation method according to claim 1, characterized in that: The pore size of the ceramic membrane in step S2 is 200-500 nm.

5. The preparation method according to claim 1, characterized in that: The mass ratio of ferric chloride, ferrous chloride and ammonia water in step T1 is 3.24:1.26:5-10, the temperature of the stirring and heating reaction is 80-90°C, the time is 3-5h, the temperature of the calcination is 400-500°C, and the time is 1-3h; the mass ratio of the magnetic particles, dopamine hydrochloride and catalyst in step T2 is 10:2-3:0.1-0.3, the catalyst is a Tris-HCl solution with a pH of 8.5-9.5, and the temperature of the heating and stirring reaction is 3 5-45°C, for 3-5h; in step T3, the molar ratio of N-methylimidazole, 3-chloropropyltrimethoxysilane and silane coupling agent KH560 is 1:1:0.2-0.3, the temperature of the heating and stirring reaction is 100-110°C, and the time is 20-24h; in step T4, the mass ratio of the fixed ionic liquid particles, modified magnetic particles and salt is 5-7:10:7-12, the stirring reaction time is 22-24h, and the salt is NaBF4 and KPF6.

6. The preparation method according to claim 1, characterized in that: In step S3, the solid-liquid ratio of the immobilized ionic liquid to the filtrate is 1:5-10 g / mL, the strength of the external magnetic field is 0.03-0.05 T, the temperature of the heating desorption is 40-50° C., and the time is 30-50 min.

7. The preparation method according to claim 1, characterized in that: The mass ratio of maca extract 1 and selenium-zinc-rich maca / zymosan protein complex in the maca extract for improving energy in step S4 is 3-5:10, and the mass ratio of maca extract 1, maca extract 2 and selenium-zinc-rich maca / zymosan protein complex in the maca extract for improving energy and sobering up is 3-5:2-4:10.

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