CN107050089A - It is a kind of to significantly increase maca composition of immunity function and preparation method thereof - Google Patents
It is a kind of to significantly increase maca composition of immunity function and preparation method thereof Download PDFInfo
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
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Abstract
The invention discloses a kind of maca composition for significantly increasing immunity function and preparation method thereof, the composition is 40 60 parts of raw material Maca extract, 20 33 parts of ginseng extract is sufficiently mixed, tabletting prepares maca composition, content >=8% of ginsenoside in polyoses content >=22% in Maca extract, ginseng extract.Preparation method includes that maca dry product is crushed, 65% 68% alcohol solution dippings, filtering, drying, crushings, water are extracted 23 times, filtering, merging filtrate, concentrates, dry, obtains maca polysaccharide extract；Ginseng is crushed, 60% 70% ethanol are extracted, filtering, merging filtrate, and the ginseng extract of 100 mesh sieves is crossed in concentration, spray drying, powder.Active constituent content is higher when the maca composition of preparation is suitable for significantly increasing the food of immunity, medicine, and processing technology is simple and easy to control, and raw material are using abundant, and production cost is low, has wide applicability in field of health care products.
Description
Technical field
The invention mainly relates to field of health care products, specifically, the present invention relates to a kind of composition skill of strengthen immunity
Art field.
Background technology
Maca (Lepidium meyenii Walp.) is Cruciferae separate row Vegetable spp one, biennial herb plant, is one
Kind of unique medicine-food two-purpose resource, originates in the plateau of more than 3000 meters of South America Peru Andes area height above sea level.Maca is obtained
The most attention of people is that in the early 1990s, researcher is found that this plant in raising property work(when finding " vigour " substitute
Remarkable efficacy on energy, makes maca turn into a nova in international health product and medicine at one stroke；And before this, work as Spain
People is when South America is captured in 1526, and the necessary article that maca is just taken root andes region by colonists as procreation of living is used
In the problem for solving people and animals' infertility, and so far still as with raising energy, fecundity effect and good tonic effect
Tribute, widely used by imperial family of Spain.Edible history of the maca in South America has had more than 5800 years, is conventionally used to strong
Body, raising fertility, improvement sexual function, antidepression, anti-anemia action etc..From early 1960s phase to the eighties, on agate
The research of coffee botany and medical value progressively systematization, some traditional roles of maca obtain Germany and North American Plant research
After the scientific validation of person, the especially eighties, the kind to maca is promoted in FAO (Food and Agriculture Organization of the United Nation) (FAO) suggestion countries in the world
Plant, chemical composition identification, active component separation and its pharmacological action of maca etc. have obtained further discussion, and research confirms agate
Coffee has effects that protrusion in terms of improving fertility, improving sexual function, antifatigue, compressive resistance, alleviation climacteric metancholia.
However, the research and application on maca bioactivity are concentrated mainly on a certain composition in its crude extract, rarely seen maca in recent years
Report with particular organisms activity.
In view of antifatigue and strengthen immunity close ties, enhancing of inventor's early stage to maca and maca composition is exempted from
Epidemic disease power effect is studied, it is found that maca can only improve mouse cell and be immunized, but can not improve mouse humoral immune, it is impossible to carry
High NK cells in mice activity, i.e. its strengthen immunity effect of exclusive use maca is not notable.
Ginseng (PanaxginsengC.A.Mey.) is the traditional rare traditional Chinese medicine of China, is consolidated with reinforcing vital energy, answering arteries and veins
Take off, reinforce the spleen to benefit the lung, blood-nourishing of promoting the production of body fluid, tranquilize the mind and promote the intelligence the effects such as.According to modern pharmacology research, ginseng has good Immune-enhancing effect
Effect, it is many-sided in reticuloendothelial system (RES), specific antibody formation, lymphocyte and thymocyte proliferation etc.
Embody.With the development of modern separate analytical technique, it is known that contained chemical composition includes ginsenoside (GS), sugar in ginseng
Class, protein, amino acid, volatile oil etc..People participates in panaxan has immune to immune organ, immunocyte and immune molecule
Adjustment effect.
At present, a kind of integration of drinking and medicinal herbs Chinese medicine ferment drink and preparation method thereof patent of invention patent (publication number
CN105995329A), herbal cuisine powder patent (publication number CN105995399A), a kind of Maca composite of regulation human-body sub-health are oral
Ginseng and maca are used to improve people as one of composition in liquid and preparation method thereof patent (publication number CN1042568218) etc.
The problems such as body immunity, inferior health, but all there is compatibility medicinal material excessively, the unclear problem of the mechanism of action, using raw material mistake
It is many, the increase of production cost can be directly resulted in, it is impossible to determine that medicinal material plays a major role, and because of feature in actual production
Increasing for raw material needs the key problem in technology point of control just to increase therewith, there is the increasing of production control difficulty or product efficacy is difficult the palm
The problem of control, extremely it is unfavorable for industrialization production and control of product quality.
Maca is employed in Maca ginseng tablet and preparation method thereof patent of invention (publication number CN103585261A) with ethanol
Solution carries out ultrasonic extraction, and ginseng, the root of kudzu vine extract through water and cross macroreticular resin respectively, prepares Maca ginseng tablet, but effect is immunized
Really undesirable, synergistic function is weak, and complex process, time-consuming longer, cost are high.A kind of Maca powder and ginseng composition and its preparation
Method patent of invention (publication number CN102613555B) uses not extracted pueraria root powder, is extracted with ginseng extract, rhodiola root
Thing, Shorthorned Epimedium P.E, Fructus lycii P.E compatibility, by controlling general ginsenoside, rhodioloside, icariin, matrimony vine thick
The content of polysaccharide etc. controls product quality and effect, quality control index showed increased, raw material, intermediate products, finished product
Detection workload is substantially increased, and equally can also increase quality control cost.CN102613555B ginseng extract weight proportions are pressed
According to the cubage of TSPG 80%, and illustrate ginseng extract according to ginsenoside content in respect of many kinds of rule of 10-80%
Lattice, after number needed for other specifications outside 80% can be converted accordingly, applied to the composition described in the invention, that is to say, that
Other low specification extract consumptions of content of ginsenoside can increase, such as using the extract of content of ginsenoside 10%, then
Its usage amount need to be increased to original 8 times, and raw material dosage is big, and not only cost is high, and drug effect is weaker, and dose is larger, inconvenience
Take.
The content of the invention
For state of the art, particularly current ginseng and maca are suitable for improving the health care such as immunity, antifatigue
Excessive using raw material during field, active constituent content is low, and immune effect is undesirable, and synergistic function is weak, complex process, consumption
When longer, production cost it is high, there is provided a kind of maca group for significantly increasing immunity function for the problem of weaker dose of drug effect is big
Compound preparation method so that processing technology is simple when ginseng and maca are suitable for the food, medicine of strengthen immunity, effectively into
Clearly demarcated true, raw material are using fully, and production cost is low, has wide applicability in field of health care products.
To achieve these goals, the technical scheme that the present invention is provided is as follows：
The present invention provides a kind of maca composition for significantly increasing immunity function, and the maca composition parts by weight are matched somebody with somebody
The raw material of ratio：40 parts -60 parts of Maca extract, 20 parts -33 parts of ginseng extract；Polyoses content >=22% in Maca extract,
Content >=8% of ginsenoside in ginseng extract.
It is preferred that, in the present invention, the raw material of the maca composition parts by weight proportioning：40 parts of Maca extract, ginseng
30 parts of extract；The content 8.7% of ginsenoside in polyoses content 23.1% in Maca extract, ginseng extract.
In the present invention, polysaccharide relative molecular weight is 5kDa-8kDa in the Maca extract.
More specifically, in the present invention, relative molecular weight 6.6kDa-6.8kDa in the polysaccharide of the Maca extract, sugared ring
Simultaneously more than 25% of polysaccharide total amount in Maca extract is accounted for comprising α types pyranose, a kind of glutinous polysaccharide of β type pyranoses.
In the present invention, the mol ratio 1 of arabinose, galactolipin and glucose in the glutinous polysaccharide of the Maca extract:
1.3:36.8。
A kind of maca composition with strengthen immunity function that the present invention is provided, preparation method is by following steps structure
Into：
(1) prepared by Maca extract：Maca dry product is crushed to particle diameter 0.2cm-0.4cm, adds 8 times of -10 times of amount 65%-
68% ethanol solution, 45 DEG C of heat-insulation soakings 2 days, filtering, filtrate is used for the immersion of next batch maca, maca dries in 50 DEG C,
Crush；It is concentration, dry by the pueraria root powder of alcohol solution dipping plus 15 times of amount water are extracted 2-3 time, filtering, merging filtrate, obtain
To maca polysaccharide extract.
(2) prepared by ginseng extract：Ginseng adds 10 times of -12 times of amount 60%-70% ethanol solutions after crushed, extracts 3
It is secondary, 1.5 hours every time, filtering, merging filtrate, 60 DEG C are concentrated into relative density 1.05, spray drying, EAT：150℃-
170 DEG C, temperature of outgoing air：75 DEG C -85 DEG C, atomisation pressure 0.4MPa is dried to moisture and is less than 5%, powder crosses 100 mesh sieves.
(3) preparation of composition：Weigh 40 parts -60 parts of the raw material Maca extract of parts by weight proportioning, ginseng extract
20 parts -33 parts are sufficiently mixed, conventionally tabletting.
More preferred, in the present invention prepared by Maca extract：Maca dry product is crushed to particle diameter 0.2cm-0.4cm, adds 8
Times 68% ethanol solution of amount, 45 DEG C of heat-insulation soakings 2 days, filtering, filtrate is used for the immersion of next batch maca, and maca is in 50 DEG C of bakings
Dry, crushing；By in the pueraria root powder of alcohol solution dipping plus 15 times amount water extract 2 times, filtering, merging filtrate, concentration, dry,
Obtain maca polysaccharide extract.
More preferred, in the present invention prepared by ginseng extract：Ginseng adds 12 times of 60% ethanol solutions of amount after crushed,
Extract 3 times, 1.5 hours every time, filter, merging filtrate, 60 DEG C are concentrated into relative density 1.05, spray drying, EAT：
160 DEG C, temperature of outgoing air：80 DEG C, atomisation pressure 0.4MPa is dried to moisture and is less than 5%, powder crosses 100 mesh sieves.
Beneficial effects of the present invention:
(1) a kind of maca composition preparation method for significantly increasing immunity function for providing of the present invention so that ginseng and
Active constituent content is higher when maca is suitable for the food, medicine of strengthen immunity, particularly by the control of technology key point
Ensure total starches content and crucial polyoses content in Maca extract, so as to ensure the strengthen immunity activity of product.The present invention
Technology key point is the preparation of maca polysaccharide extract, and processing technology is while ensureing to obtain high level polyoses extract
Again simple and easy to control, raw material are using fully, and production cost is low, has wide applicability in field of health care products.
(2) polysaccharide feature is in Maca extract of the present invention, relative molecular weight 5-8kDa, and with relative molecular weight
6.6kDa-6.8kDa, the mol ratio 1 of arabinose, galactolipin and glucose:1.3:36.8, sugared ring is simultaneously comprising α type pyrans
Based on sugar, a kind of glutinous polysaccharide of β type pyranoses, more than 25% of polysaccharide total amount in extract is at least accounted for.Pharmacological testing proves, agate
Polysaccharide relative molecular weight concentrates on 5-8kDa in coffee extract, and above-mentioned glutinous polyoses content is higher, then Maca extract and people
The combination microbic activity of conopsea extraction is stronger, therefore, and such polysaccharide is to improve present composition strengthen immunity activity
Key component.
(3) research on maca polysaccharide is concentrated mainly on neutral polysaccharide and acidic polysaccharose, and grinding for polysaccharide is sticked on maca
Study carefully report compare it is less.Find that above-mentioned glutinous polysaccharide is exempted to the enhancing for lifting composition during the present composition is prepared
Epidemic disease power effect plays an important roll, and this is also the discovery first in this kind of glutinous polysaccharide researches, and the discovery make it that ginseng and maca are suitable
Processing technology is simple when in the food, medicine for strengthen immunity, and clearly, raw material are produced into active ingredient using abundant
This is low.
(4) present composition group has good humoral immune function (mouse antibodies cellulation, mouse half hemolysis
Value) (P<0.05), with certain cellular immune function (influence to mouse delayed allergy ability) (P<0.01), and
And NK cytoactives (P can be significantly improved<0.01), i.e., the present composition has strengthen immunity function, and the present invention passes through
The Maca extract of ad hoc fashion prepares and realizes synergistic function, the increasing of composition with the reasonable compatibility of ginseng extract
Strong efficacy of immunity is significantly improved：Research has shown that, maca can only improve mouse cell and be immunized, but can not improve mouse body fluid and exempt from
Epidemic disease, it is impossible to improve NK cells in mice activity, the present invention makes group by the preparation of high content maca polysaccharide extract with reasonable compatibility
Compound has actively impact to mouse humoral immune, cellular immunity, NK cytoactives, with higher application value.
Brief description of the drawings
Fig. 1 is shown as the glutinous polysaccharide HPIC spectrograms of the present invention.
Fig. 2 is shown as the glutinous polysaccharide FT-IR spectrograms of the present invention.
In Fig. 1,1- arabinoses, 2- galactolipins, 3- glucose.
Embodiment
The embodiment to the present invention is described in further detail below, but the method for the present invention is not limited to following realities
Apply example.
In the present invention, used ginseng, maca, Kunming mouse, SPF Healthy females mouse, multi-function microplate reader, dioxy
Change carbon incubator, T1000 types electronic balance, JA2003 types electronic balance, JJ-100 electronic balances, syklofosfamid ampoule,
RPMI1640, YAC-1 cell, LDH matrix liquids, SG-603 Biohazard Safety Equipments, SRBC, SA buffer solution, centrifuge, spectrophotometric
Meter, water-bath, vortex mixer, absolute ethyl alcohol, glucose etc. are bought using market public channel and obtained.
Embodiment one：Significantly increase the maca composition of immunity function
The present invention provides kind of the maca composition with strengthen immunity function, the maca composition parts by weight proportioning
Raw material：40 parts -60 parts of Maca extract, 20 parts -33 parts of ginseng extract；Polyoses content >=22% in Maca extract, people
Content >=8% of ginsenoside in conopsea extraction.
Embodiment two：Significantly increase the maca composition of immunity function
In the present invention, the raw material of the maca composition parts by weight proportioning：40 parts of Maca extract, ginseng extract 30
Part；The content 8.7% of ginsenoside in polyoses content 23.1% in Maca extract, ginseng extract.
In the present invention, polysaccharide relative molecular weight is 5-8kDa in the Maca extract.
More specifically, in the present invention, relative molecular weight 6.6kDa-6.8kDa in the polysaccharide of the Maca extract, sugared ring
Simultaneously more than 25% of polysaccharide total amount in Maca extract is accounted for comprising α types pyranose, a kind of glutinous polysaccharide of β type pyranoses.
In the present invention, the mol ratio 1 of arabinose, galactolipin and glucose in the glutinous polysaccharide of the Maca extract:
1.3:36.8。
Embodiment three：Significantly increase the maca composition preparation method of immunity function
The invention provides a kind of maca composition preparation method with strengthen immunity function, by following steps structure
Into：
(1) prepared by Maca extract：Maca dry product is crushed to particle diameter 0.2cm-0.4cm, adds 8 times of -10 times of amount 65%-
68% ethanol solution, 45 DEG C of heat-insulation soakings 2 days, filtering, filtrate is used for the immersion of next batch maca, maca dries in 50 DEG C,
Crush；It is concentration, dry by the pueraria root powder of alcohol solution dipping plus 15 times of amount water are extracted 2-3 time, filtering, merging filtrate, obtain
To maca polysaccharide extract.
(2) prepared by ginseng extract：Ginseng adds 10 times of -12 times of amount 60%-70% ethanol solutions after crushed, extracts 3
It is secondary, 1.5 hours every time, filtering, merging filtrate, 60 DEG C are concentrated into relative density 1.05, spray drying, EAT：150℃-
170 DEG C, temperature of outgoing air：75 DEG C -85 DEG C, atomisation pressure 0.4MPa is dried to moisture and is less than 5%, powder crosses 100 mesh sieves.
(3) preparation of composition：Weigh 40-60 parts of the raw material Maca extract of parts by weight proportioning, ginseng extract 20-
33 parts are sufficiently mixed, tabletting.
Example IV：The preparation of Maca extract
The preparation of Maca extract：Maca dry product is crushed to particle diameter 0.2cm-0.4cm, adds 8 times of 68% ethanol solutions of amount,
45 DEG C of heat-insulation soakings 2 days, filtering, filtrate is used for the immersion of next batch maca, maca in 50 DEG C of drying, crush；By ethanol
Add 15 times of amount water to extract according to a conventional method in the pueraria root powder of solution immersion 2 times, filter, merging filtrate, concentrate, dry, obtain agate
Coffee polyoses extract.
Embodiment five：Polyoses content is detected in Maca extract
1 method
1.1 total starches assays
Press《Health food functional component detection method》" the phenolsulfuric acid spectrophotometric of Thick many candies is surveyed in (Bai Hong chief editors)
Determine method " in method detection.
1.2 significant Analysis of polysaccharides
Sample treatment：2M TFA, 110 DEG C of hydrolysis 6h are added in maca polysaccharide, is concentrated in vacuo, removes acid.
HPIC conditions：CarboPac PA20 posts (3 × 150mm), 0.5mL/min, water (A), 250mM NaOH (B), 1M
NaAc (C) gradient elution.
2 results
2.1 total starches contents
It is 25.9% to measure polyoses content in Maca extract according to example IV method.
2.2 significant Analysis of polysaccharides
A kind of glutinous polysaccharide that HPIC analyses are obtained in maca polysaccharide extract, according to collection of illustrative plates ordinate abundance
Abundance understands that the mol ratio of glutinous the polysaccharide arabinose (1), galactolipin (2) and glucose (3) is 1:1.3:36.8, ginseng
See Figure of description 1, sugared ring is simultaneously comprising α types pyranose, β type pyranoses, referring to Figure of description 2.
Embodiment six：The preparation of Maca extract
The preparation of Maca extract：Maca dry product is crushed to particle diameter 0.2cm-0.4cm, adds 10 times of 65% ethanol of amount molten
Liquid, 45 DEG C of heat-insulation soakings 2 days, filtering, filtrate is used for the immersion of next batch maca, maca in 50 DEG C of drying, crush；By second
Add 15 times of amount water to extract according to a conventional method in the pueraria root powder of alcoholic solution immersion 3 times, filter, merging filtrate, concentrate, dry, obtain
Maca polysaccharide extract.After testing, polyoses content is 22.2% in the extract.
Embodiment seven：The preparation of ginseng extract
Ginseng adds 12 times of 60% ethanol solutions of amount after crushed in the present invention, extracts 3 times, 1.5 hours every time, filters,
Merging filtrate, 60 DEG C are concentrated into relative density 1.05, spray drying, EAT：160 DEG C, temperature of outgoing air：80 DEG C, spraying pressure
Power 0.4MPa, dries to moisture and is less than 5%, powder crosses 100 mesh sieves.After testing, content of ginsenoside is 8.7%.
Embodiment eight：Strengthen immunity check experiment
1 material and method
1.1 animals and packet
Kunming mouse 70, every group 10, carries out immunosuppressed mice and organ index measure examination is immunized by random point 7 groups
Test.SPF Healthy females mouse 60, every group 10, carries out NK cell activity assays by random point 6 groups.
1.2 dosage choices give mode with tested material
If 1 group of composition, 40 parts of Maca extract, 30 parts of ginseng extract is sufficiently mixed (preparation method preparation of the present invention
Composition, Maca extract polyoses content 23.1%, ginseng extract content of ginsenoside 8.70%).
40 parts of 2 groups of maca water extracts of composition (directly being extracted without ethanol immersion with water), 30 parts of ginseng extract fills
Divide mixing (maca water extract polyoses content 10.3%, ginseng alcohol extract content of ginsenoside 8.70%).
3 groups of composition, 40 parts of maca ethanol extract, 30 parts of ginseng extract is sufficiently mixed (ginseng alcohol extract ginsenoside
Content 8.70%).
Maca extract group (maca polysaccharide content 23.1%).
Ginseng extract group (ginseng water extract content of ginsenoside 6.21%).
Each component test dose is 0.60g/kgbw/d, and blank control group replaces tested material with sterilized water.Gavage amount
0.2ml/10g·bw。
1.3 instruments and reagent
Electronic balance, multi-function microplate reader, CO2gas incubator, JJ-100 electronic balances.Syklofosfamid ampoule,
RPMI1640, YAC-1 cell, LDH matrix liquids.
1.4 test method
1.4.1 organ index is immunized in immunosuppressed mice
Kunming mouse is grouped gastric infusion, continuous gavage 14d.11d is tested, in addition to blank control group, each group mouse
Ring phosphorus gland amine is injected intraperitoneally with 50mg/kg respectively, continuous 3d causes animal immune to suppress model, blank control group intraperitoneal injection
Normal saline.After 14d administrations 1h, mouse is put to death in dislocation, is weighed, and takes spleen, thymus gland, claims weight in wet base, calculates spleen index
And thymus index.
Spleen index=spleen weight/body weight (mg/g)；
Thymus index=chest gland weight/body weight (mg/g).
1.4.2NK the measure of cytoactive
Test mice cervical dislocation is put to death, sterile to take spleen, and splenocyte suspension 2 × 10 is made7This is effector cell to individual/mL,
It is 4 × 10 to take the well-grown YAC-1 cells RPMI1640 complete culture solutions adjustment cell concentrations of 24h after passage5Individual/mL this
For target cell；Target cell and each 100 μ L of effector cell (effect target is than 50: 1) are taken, is added in U-shaped 96 well culture plate；Target cell is natural
Release aperture adds target cell and each 100 μ L of nutrient solution, and target cell maximum release aperture adds target cell and each 100 μ L of 2.5%Triton；On
State items and be all provided with 3 parallel holes, in 37 DEG C, 5%CO24h is cultivated in incubator, then by 96 well culture plates with 1500r/min from
Heart 5min, draws per hole in the well culture plate of supernatant 100 μ L horizontalizations bottom 96, while LDH matrix liquids 100 μ L are added, it is anti-according to room temperature
3-10min is answered, 1mol/L HCl30 μ L are added per hole, optical density (OD) is determined at ELIASA 490nm, NK cells is calculated and lives
Property.
NK cytoactives=(reacting hole OD values-Spontaneous release hole OD values)/(maximum release aperture OD values-Spontaneous release hole OD
Value) × 100.
2 results
The influence of organ index is immunized in 2.1 pairs of immunosuppressed mices
Intraperitoneal injection 50mg/kg endoxan can significantly reduce the index and spleen index of mouse, thymus index；Compared to model comparison
Group, composition 1 (present composition) is remarkably improved mouse spleen index, thymus index (P<0.01) (P is significantly improved<
0.01), composition 3 (maca ethanol extract+ginseng extract) significantly improves mouse spleen index, thymus index (P<
0.05), composition 2 (maca water extract (directly extracts)+ginseng extract) only improves mouse thymus index (P<0.05), maca
Extract only improves mouse spleen index (P<0.05), and ginseng extract to the two all without remarkable effect (P>0.05).Group
The influence that compound 1 (present composition) immunosuppressed mice is immunized organ index is maximum.
Table 1：Sample immunosuppressed mice is immunized the influence (n=10) of organ index
| Group | Index and spleen index (mg/g) | Thymus index (mg/g) |
| Blank control group | 5.1±0.3 | 4.2±0.4 |
| Model control group | 1.8±0.2 | 1.2±0.4 |
| 1 group of composition | 2.7±0.3** | 1.8±0.4** |
| 2 groups of composition | 2.0±0.3 | 1.4±0.3* |
| 3 groups of composition | 2.1±0.3* | 1.4±0.3* |
| Maca extract group | 2.1±0.4* | 1.2±0.2 |
| Ginseng extract group | 1.9±0.3 | 1.2±0.5 |
The influence of 2.2 pairs of NK cells in mice activity
Composition 1 (present composition) significantly improves NK cells in mice activity (P<0.05), composition 2, composition 3,
Maca extract, ginseng extract on NK cells in mice activity there are no significant influence (P>0.05).
Table 2：Influence (n=10) of each group to NK cells in mice activity
| Group | NK cytoactives (%) |
| Blank control group | 30.12±5.72 |
| 1 group of composition | 38.54±5.64* |
| 2 groups of composition | 33.21±5.37 |
| 3 groups of composition | 34.14±4.27 |
| Maca extract group | 34.97±5.42 |
| Ginseng extract group | 32.77±5.23 |
Composition 1 (present composition) can significantly improve hypoimmunity mouse immune internal organs (spleen, thymus gland) index
(P<0.01) NK cytoactives (P, is significantly improved<0.05), effect is substantially better than composition 2, composition 3, Maca extract, people
Conopsea extraction.The present composition is combined by the preparation of high content maca polysaccharide extract with ginseng extract realizes combination
Thing strengthen immunity is active to be significantly improved.
Embodiment eight：Maca composition significantly increases immunity function evaluation
1 material and method
1.1 animals and packet
SPF Healthy female mouse, are randomly divided into three big groups by body weight and carry out pedal swelling experiment, the examination of NK cytoactives respectively
Test, antibody-producting cell detection and half hemolytic value HC50Determination test.
1.2 dosage choices give mode with tested material
If the basic, normal, high dosage group of the present composition be respectively 0.30g/kgbw/d, 0.60g/kgbw/d,
1.80g/kgbw/d, 0g/kgbw/d group replace tested material, gavage amount 0.2ml/10gbw, continuous gavage with sterilized water
30d。
1.3 instruments and reagent
T1000 types electronic balance, JA2003 types electronic balance, SG-603 Biohazard Safety Equipments, multi-function microplate reader, titanium dioxide
Carbon incubator.SRBC, RPMI1640, SA buffer solution, YAC-1 cells, LDH matrix liquids etc..
1.4 test method
1.4.1 delayed allergy (DTH) (vola pedis thickens method)
Mouse peritoneal injection 2% (v/v) sheep red blood cell (SRBC) suspension sensitization, measures left back foot plantar thickness, in measuring point
Left back foot plantar thickness is measured after 20% (v/v) sheep red blood cell (SRBC) suspension (20ul is per mouse), 24h is subcutaneously injected.
The preceding vola pedis thickness (mm) of vola pedis thickness (mm)-injection after swelling degree of the paw (mm)=injection after injection.
1.4.2 antibody-producting cell detection (Jerne improves slide methods)
0.2mL2% (v/v) sheep red blood cell (SRBC) suspension is injected intraperitoneally per mouse.Mouse is put to death after 4d, spleen is taken, splenocyte is made
Suspension.Hemolysis plaque number is counted by Jerne improves slide methods,
Hemolysis plaque number=hemolysis plaque counts × 320.
1.4.3HC50 determine
Per mouse, intraperitoneal injection 0.2ml2% sheep red blood cell (SRBC) suspensions are immunized.Eyeball is extractd after 4d and takes blood, centrifugation, collection
Serum.It is molten by OD values when method of inspection determination sample pipe and SRBC half hemolysis with SA buffer solutions by 200 times of serum-dilution
The amount of sanguinin is with half hemolytic value (HC50) represent.
HC50OD value × extension rate during=sample OD value/SRBC half hemolysis.
1.4.4NK the measure of cytoactive
Test mice cervical dislocation is put to death, sterile to take spleen, and splenocyte suspension 2 × 10 is made7This is effector cell to individual/mL,
It is 4 × 10 to take the well-grown YAC-1 cells RPMI1640 complete culture solutions adjustment cell concentrations of 24h after passage5Individual/mL this
For target cell；Target cell and each 100 μ L of effector cell (effect target is than 50: 1) are taken, is added in U-shaped 96 well culture plate；Target cell is natural
Release aperture adds target cell and each 100 μ L of nutrient solution, and target cell maximum release aperture adds target cell and each 100 μ L of 2.5%Triton；On
State items and be all provided with 3 parallel holes, in 37 DEG C, 5%CO24h is cultivated in incubator, then by 96 well culture plates with 1500r/min from
Heart 5min, draws per hole in the well culture plate of supernatant 100 μ L horizontalizations bottom 96, while LDH matrix liquids 100 μ L are added, it is anti-according to room temperature
3-10min is answered, 1mol/L HCl30 μ L are added per hole, optical density (OD) is determined at ELIASA 490nm, NK cells is calculated and lives
Property.
NK cytoactives=(reacting hole OD values-Spontaneous release hole OD values)/(maximum release aperture OD values-Spontaneous release hole OD
Value) × 100.
2 results
The influence of 2.1 pairs of mouse DTHs
Basic, normal, high dosage group mouse delayed allergy ability is compared with control group to be improved, and middle dose group is improved
With statistical significance (P<0.05), high dose group is then significantly increased (P<0.01).
Table 3：Influence (n=10) of each group to mouse DTH
| Group | Swelling degree of the paw (mm) |
| Solvent control group | 0.26±0.13 |
| Low dose group | 0.33±0.19 |
| Middle dose group | 0.46±0.19 |
| High dose group | 0.51±0.14** |
The influence of 2.2 pairs of mouse antibodies cellulations
Basic, normal, high dosage group mouse antibodies cellulation number is improved, and high dose group is compared with control group to be significantly improved
(P<0.05)。
Table 4：Each group generates the influence (n=10) of number to antibody cell
2.3 couples of mouse HC50Influence
Basic, normal, high dosage group half hemolytic value is improved, and wherein high dose group half hemolytic value is compared aobvious with control group
Write and improve (P<0.05).
Table 5：Each group is to mouse HC50Influence (n=10)
| Group | HC50 |
| Solvent control group | 153.12±32.24 |
| Low dose group | 169.24±38.81 |
| Middle dose group | 182.26±34.26 |
| High dose group | 202.22±43.42* |
The influence of 2.4 pairs of NK cells in mice activity
Basic, normal, high dosage group NK cytoactives are all improved, and wherein high dose group NK cytoactives are apparently higher than solvent pair
According to group (P<0.01).
Table 6：Influence (n=10) of each group to NK cells in mice activity
Present composition group has good humoral immune function (mouse antibodies cellulation, mouse half hemolytic value)
(P<0.05), with certain cellular immune function (influence to mouse delayed allergy ability) (P<0.01), and
NK cytoactives (P can be significantly improved<0.01), i.e., the present composition has strengthen immunity function, and the present invention passes through spy
The Maca extract for determining mode prepares and realizes synergistic function, the enhancing of composition with the reasonable compatibility of ginseng extract
Efficacy of immunity is significantly improved：Maca can only improve mouse cell and be immunized, but can not improve mouse humoral immune, it is impossible to improve small
Mouse NK cytoactives, the preparation of the maca polysaccharide extract for the relative molecular weight 5-8kDa that the present invention passes through high content is with rationally matching somebody with somebody
5 make composition have actively impact to mouse humoral immune, cellular immunity, NK cytoactives, with higher application value.Especially
Maca polysaccharide relative molecular weight 6.6kDa-6.8kDa in its Maca extract of the present invention, arabinose, galactolipin and glucose
Mol ratio 1:1.3:36.8, sugared ring comprising α types pyranose, based on a kind of glutinous polysaccharide of β type pyranoses, at least accounts for extract simultaneously
More than the 25% of middle polysaccharide total amount, then the combination microbic activity of Maca extract and ginseng extract is stronger, and therefore, such is more
Sugar is the key component for improving present composition strengthen immunity activity.Pharmacological testing proves, polysaccharide phase in Maca extract
5-8kDa is concentrated on to molecular weight, and above-mentioned glutinous polyoses content is higher to mouse humoral immune, cellular immunity, NK cytoactives
Improve more obvious.
As described above, you can preferably realize the present invention, the above embodiments are only the side of being preferable to carry out to the present invention
Formula is described, and not the scope of the present invention is defined, and on the premise of design spirit of the present invention is not departed from, this area is general
Various modifications and improvement that logical technical staff makes to technical scheme, all should fall into present invention determine that protection domain
It is interior.
Claims (8)
1. a kind of maca composition with strengthen immunity function, it is characterised in that the maca composition parts by weight are matched somebody with somebody
The raw material of ratio：40 parts -60 parts of Maca extract, 20 parts -33 parts of ginseng extract；Polyoses content >=22% in Maca extract, people
Content >=8% of ginsenoside in conopsea extraction.
2. a kind of maca composition with strengthen immunity function as claimed in claim 1, it is characterised in that the maca
The raw material of composition weight number proportioning：40 parts of Maca extract, 30 parts of ginseng extract；Polyoses content in Maca extract
23.1%, the content 8.7% of ginsenoside in ginseng extract.
3. a kind of maca composition with strengthen immunity function as claimed in claim 1, it is characterised in that the maca
Polysaccharide relative molecular weight is 5-8kDa in extract.
4. a kind of maca composition with strengthen immunity function as claimed in claim 3, it is characterised in that the maca
Relative molecular weight 6.6kDa-6.8kDa in the polysaccharide of extract, the one kind of sugared ring simultaneously comprising α types pyranose, β type pyranoses is sticked
Polysaccharide accounts for more than 25% of polysaccharide total amount in Maca extract.
5. a kind of maca composition with strengthen immunity function as claimed in claim 4, it is characterised in that the maca
The mol ratio 1 of arabinose, galactolipin and glucose in the glutinous polysaccharide of extract:1.3:36.8.
6. a kind of preparation method of the maca composition with strengthen immunity function, it is characterised in that be made up of following steps：
(1) prepared by Maca extract：Maca dry product is crushed to particle diameter 0.2cm-0.4cm, adds 8 times of -10 times of amount 65%-68% ethanol
Solution, 45 DEG C of heat-insulation soakings 2 days, filtering, filtrate is used for the immersion of next batch maca, maca in 50 DEG C of drying, crush；By
It is concentration, dry in the pueraria root powder of alcohol solution dipping plus 15 times of amount water are extracted 2-3 time, filtering, merging filtrate, obtain maca many
Sugar extract；
(2) prepared by ginseng extract：Ginseng add after crushed 10 times -12 times amount 60%-70% ethanol solutions, extract 3 times, every time
1.5 hours, filtering, merging filtrate, 60 DEG C are concentrated into relative density 1.05, spray drying, EAT：150 DEG C -170 DEG C, row
Air temperature：75 DEG C -85 DEG C, atomisation pressure 0.4MPa is dried to moisture and is less than 5%, powder crosses 100 mesh sieves；
(3) preparation of composition：Weigh parts by weight proportioning 40 parts -60 parts of raw material Maca extract, 20 parts of ginseng extract -
33 parts are sufficiently mixed, conventionally tabletting.
7. a kind of preparation method of the maca composition with strengthen immunity function as claimed in claim 6, its feature exists
In Maca extract is prepared as：Maca dry product is crushed to particle diameter 0.2cm-0.4cm, adds 8 times of 68% ethanol solutions of amount, 45 DEG C of guarantors
Temperature immersion 2 days, filtering, filtrate is used for the immersion of next batch maca, and maca is dried in 50 DEG C, crushed；Soaked by ethanol solution
In the pueraria root powder of bubble plus 15 times amount water extract 2 times, filtering, merging filtrate, concentration, dry, obtain maca polysaccharide extract.
8. a kind of preparation method of the maca composition with strengthen immunity function as claimed in claim 6, its feature exists
In the ginseng extract is prepared as：Ginseng adds 12 times of 60% ethanol solutions of amount after crushed, extracts 3 times, 1.5 is small every time
When, filtering, merging filtrate, 60 DEG C are concentrated into relative density 1.05, spray drying, EAT：160 DEG C, temperature of outgoing air：80
DEG C, atomisation pressure 0.4MPa is dried to moisture and is less than 5%, powder crosses 100 mesh sieves.
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