CN108741056A - A kind of preparation method for climbing sandworm extract adjusting immune function - Google Patents
A kind of preparation method for climbing sandworm extract adjusting immune function Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to functional food preparation field, specially a kind of preparation method for climbing sandworm extract adjusting immune function.The method comprising the steps of 1)The fresh and alive sandworm that climbs is cleaned with pure water, is removed end to end and internal organ, is cleaned and drain, it is spare in -80 DEG C of quick freezes;2)Before active constituent extraction, with the fritter for being 5mm is crushed after 25 DEG C of warm water constant-temperature thawings, 40 mesh sieve crushed again after 36h is freeze-dried in freeze drier, it is spare at a temperature of being sealed in -20 DEG C;3)By solid-liquid ratio 1:2-1:Distilled water is added in 4 ratio, using 25 DEG C of warm water immersion, is extracted and is filtered with ultrasonic wave drug processor, take filtrate, and upper filtrate, which is placed in freeze drier, to be dried 36 hours, collects the sample after drying to obtain the final product.The obtained protection climbed sandworm extract and hypoimmunity mouse cell is immunized;Climb sandworm water-soluble extractive has significant protective effect to the specific humoral immunity of hypoimmunity mouse.
Description
Technical field
The invention belongs to functional food preparation field, specially a kind of preparation for climbing sandworm extract adjusting immune function
Method.
Background technology
It is the traditional rare dietotherapy invigorant of Chinese Minority Nationalities to climb sandworm, and element is eponymous with ginseng, pilose antler, and tradition is few
The good medicine that number ethno-medicine is praised highly.It climbs in sandworm body that protein content is very high in dry matter, is 67.69%;Fat content is relatively low,
It is 10.4%;Total sugar content is very low, is 1.59%;Content of ashes is 7.05%.Modern Chinese medicine medical research is thought, sandworm is climbed
It is sweet in flavor and warm in property, enter kidney lung channel, has that tonifying lung kidney, cough-relieving are coughed, benefit is deficient, helps the effect of vital essence, be suitable for deficiency of both the lung and kidney, vital essence not
The diseases such as foot, impotence and seminal emission, cough lose heart, spontaneous sweating, soreness and weakness of waist and knees.Again because its mild-natured power is slow, the flat tonifying yin sun of energy, is person in middle and old age
Body declines, is weak after being ill, postpartum weakness person tune tonic eats good merchantable brand.But it is seldom to climbing the system research of sandworm both at home and abroad at present, all
It is only simple to start in terms of ingredient and living environment, do not enter in terms of edible, medical value and its pharmacology really
Hand, do not accomplish food be used as medicine it is reasonable can be according to;To also not explored further in terms of climbing the extraction of sandworm effective component.
Climb sandworm and be known as animal ginseng again, in vivo the utility containing human needs, be it is a kind of not by
It pollutes, derived from environmentally protective biological food.Civil, climb sandworm people be exactly in the eyes a lot of cuisines, mainly exist
Restaurant occurs, and there are no any to climb sandworm converted products on the market.It is very few to the utilization of its active constituent research at present,
And civil edible also its effect science is proved without pharmacological experiment data.It has had not yet to see document or patent report is climbed
Sandworm adjusts the report of body's immunity.
Invention content
Present invention aims at for the above technical problem, provide a kind of a kind of adjusting enhancing body's immunity to exempt from
The preparation method for climbing sandworm extract of epidemic disease function.This method elaborates the extractive technique for climbing sandworm extract, fills up
The blank in this field;In addition the application provides the mechanism for climbing sandworm extract enhancing body's immunity the reality of science
Test Datum Argument;To climb the deep development application fetches new direction of sandworm, also provided to climb the development and application of sandworm medical value
Experimental data supports, and a piece of new world is opened up in the development and application to climb sandworm.
To achieve the goals above, the present invention adopts the following technical scheme that:
A kind of preparation method for climbing sandworm extract adjusting immune function, includes the following steps:
1) preparation of sandworm coarse powder is climbed:The fresh and alive sandworm that climbs is cleaned with pure water, removes end to end and internal organ, cleans drip
It is dry, it is freezed in -80 DEG C rapid (adfreezing in 20 minutes) spare;
2) before active constituent extraction, with 25 DEG C of warm water constant-temperature thawings, with blender by its broken fritter for 5mm, Yu Leng
It is freeze-dried 36h in lyophilizer (temperature is subzero 80 DEG C);Dry polypide was broken into 40 mesh sieve, sealing with pulverizer
It is spare at a temperature of -20 DEG C;
3) extraction of sandworm extract is climbed:By solid-liquid ratio 1:2-1:Distilled water is added in 4 ratio, is soaked using 25 DEG C of warm water
Steep half an hour, with ultrasonic wave drug processor extract, filtering, take filtrate, by upper filtrate be placed in freeze drier (temperature be-
80 DEG C) it is 36 hours dry, the sample after drying is collected to get sandworm extract is climbed.
A method of it adjusting immune function and climbs sandworm skimmed milk, include the following steps:
Step 1) and step 2) are the same as the step 1) and step in the preparation method for climbing sandworm extract for adjusting immune function
2)。
Step 3 '), step 2) is finally obtained into spare substance according to solid-liquid ratio 1:5-1:Petroleum ether is added in 10 ratio,
After 20-40 DEG C of water-bath flows back 2h, petroleum ether is recovered by filtration to get sandworm skimmed milk is climbed.
A kind of preparation process for adjusting immune function and climbing sandworm water extract coarse powder, includes the following steps:Take preceding method
What is be prepared climbs sandworm skimmed milk, is 1 according to solid-liquid ratio:4-1:Pure water is added in 20 ratio, is carried using ultrasonic wave auxiliary
It follows the example of, 15-60 DEG C of Extracting temperature, extraction time 15-60min, is collected by filtration filtrate, then with the filtering with microporous membrane of 0.45nm,
Filtrate is collected, is freeze-dried in freeze drier to get sandworm water extract coarse powder is climbed.
Compared with prior art, the beneficial effects of the invention are as follows:
(1), the application, which is directed to, climbs the extraction process of sandworm water solubility effective component and does system research, investigate extraction time,
The factors such as temperature, solid-liquid ratio illustrate the optimum process for climbing the extraction of sandworm active constituent to the factor of extraction yield;It can obtain in this way
To highest protein content.
(2), influence of the sandworm water-soluble extractive to body's immunity is climbed in the application evaluation, has been inquired into and has been climbed sandworm and carry
Object is taken to improve the mechanism of immunity of organism;
(3), the application, which illustrates, climbs sandworm extract extractive technique, has filled up the blank in this field;Separately
The experimental data that outer the application provides the mechanism for climbing sandworm extract enhancing body's immunity science is proved;To climb sandworm
Deep development application fetches new direction, also for climb the development and application of sandworm medical value provide experimental data support, to climb sand
A piece of new world is opened up in the development and application of worm.
(4) protection climbed sandworm extract and hypoimmunity mouse cell is immunized that this method is prepared is used;It climbs
Sandworm water-soluble extractive has significant protective effect to the specific humoral immunity of hypoimmunity mouse.
Specific implementation mode
Below by embodiment, the present invention is described in further detail, but protection scope of the present invention is not limited to
This specific implementation mode.The present invention expands to any new feature disclosed in the present specification or any new combination, and drapes over one's shoulders
The step of any new method or process of dew or any new combination.
Embodiment 1:
A kind of preparation method for climbing sandworm extract adjusting immune function, includes the following steps:
1) preparation of sandworm coarse powder is climbed:The fresh and alive sandworm that climbs is cleaned with pure water, removes end to end and internal organ, cleans drip
It is dry, it is freezed in -80 DEG C rapid (adfreezing in 20 minutes) spare;
2) before active constituent extraction, with 25 DEG C of warm water constant-temperature thawings, with blender by its broken fritter for 5mm, in -80
DEG C freeze drier in be freeze-dried 36h;Dry polypide was broken into 40 mesh sieve with pulverizer, is sealed in -20 DEG C of temperature
It is spare under degree;
3) extraction of sandworm extract is climbed:It is 1 by material g liquid ml ratios:Distilled water is added in 2 ratio, using 25 DEG C of warm water
Half an hour is impregnated, is extracted with ultrasonic wave drug processor, filtering takes filtrate, upper filtrate is placed in -80 DEG C of freeze drier
It is 36 hours dry, the sample after drying is collected to get sandworm extract is climbed.
Embodiment 2:
A method of it adjusting immune function and climbs sandworm skimmed milk, include the following steps:
1) preparation of sandworm coarse powder is climbed:The fresh and alive sandworm that climbs is cleaned with pure water, removes end to end and internal organ, cleans drip
It is dry, it is freezed in -80 DEG C rapid (adfreezing in 20 minutes) spare;
2) before active constituent extraction, with 25 DEG C of warm water constant-temperature thawings, with blender by its broken fritter for 5mm, in -80
DEG C freeze drier in be freeze-dried 36h;Dry polypide was broken into 40 mesh sieve with pulverizer, is sealed in -20 DEG C of temperature
It is spare under degree;
3) step 2), is finally obtained into spare substance according to solid-liquid ratio 1:5-1:Petroleum ether is added in 10 ratio, in 20-
After 40 DEG C of water-bath reflux 2h, petroleum ether is recovered by filtration to get sandworm skimmed milk is climbed.
Embodiment 3:
A kind of preparation process for adjusting immune function and climbing sandworm water extract coarse powder, includes the following steps:
1) preparation of sandworm coarse powder is climbed:The fresh and alive sandworm that climbs is cleaned with pure water, removes end to end and internal organ, cleans drip
It is dry, it is freezed in -80 DEG C rapid (adfreezing in 20 minutes) spare;
2) before active constituent extraction, with 25 DEG C of warm water constant-temperature thawings, with blender by its broken fritter for 5mm, in -80
DEG C freeze drier in be freeze-dried 36h;Dry polypide was broken into 40 mesh sieve with pulverizer, is sealed in -20 DEG C of temperature
It is spare under degree;
3) step 2), is finally obtained into spare substance according to solid-liquid ratio 1:5-1:Petroleum ether is added in 10 ratio, in 20-
After 40 DEG C of water-bath reflux 2h, petroleum ether is recovered by filtration to get sandworm skimmed milk is climbed;
4) it takes and climbs sandworm skimmed milk, be 1 according to material g liquid ml ratios:4-1:Pure water is added in 20 ratio, auxiliary using ultrasonic wave
Help extraction method, 40 DEG C, extraction time 45min of Extracting temperature is collected by filtration filtrate, then with the filtering with microporous membrane of 0.45nm, receives
Collect filtrate, is freeze-dried in freeze drier to get sandworm water extract coarse powder is climbed.
Experiment 1:
By taking the method and step in embodiment 1 as an example, influence result of the test ultrasonic time to extraction yield.
Under the conditions of 40 DEG C of ultrasonic temperature, material g liquid ml ratios are 1:5, it is obtained a result with the variation of extraction time and is shown in Table 1:It carries
Ascendant trend is presented in the protein content taken as time increases, and after rising to 45min at any time, protein content is not further added by,
It is maintained within the scope of one and changes less.The result shows that:In 45min, total nitrogen content reaches best, be 294.5mg/g, from when
Between upper consider that 45min is optimum extraction process.
Influence of 1 ultrasonic time of table to albumen yield
Group number | 1 | 2 | 3 | 4 |
Ultrasonic time/min | 15 | 30 | 45 | 60 |
Protein content mg/g | 234.3 | 225.5 | 294.5 | 290.8 |
Experiment 2:
By taking the method and step in embodiment 1 as an example, influence result of the Test extraction temperature to yield
Under the conditions of ultrasonic time 45min, solid-liquid ratio 1:5, show that experimental result is shown in Table 2 using temperature as variable quantity:Extraction
Protein content ascendant trend is presented as the temperature increases, with temperature rise to 45 DEG C, albumen yield reaches maximum value, temperature
Continue to rise the decline of albumen yield, it may be related to protein structure destruction with stable raising.
Influence of 2 ultrasonic temperature of table to albumen yield
Experiment 3:
By taking the method and step in embodiment 1 as an example, test material g liquid ml compares the influence result of extraction yield.
Under the conditions of ultrasonic time 45min, at 45 DEG C, the protein content of extraction changes with solid-liquid ratio and is become for temperature control
Change, the results are shown in Table 3:When solid-liquid ratio is 1:When 10, albumen yield reaches maximum value.The result shows that:Ultrasonic 45min, temperature 45 C,
Solid-liquid ratio 1:When 10, albumen yield reaches maximum value, is 224.42mg/g.The best solid-liquid ratio of extraction process is 1:10.
Influence of 3 solid-liquid ratio of table to albumen yield
Experiment 4:
The Optimal Experimental of sandworm extract extraction conditions is climbed, by taking embodiment 1 as an example.
The factor that response (total nitrogen content) is had a significant impact in selection experiment of single factor.Using Design-
Expert8.0 carries out regression analysis to experimental data.It is determined that factor level range is by experiment of single factor:Ultrasonic time is
30-60min;Ultrasonic temperature is 30-60 DEG C;Solid-liquid ratio is 1:15-1:5.
The basic, normal, high experimental level of each independent variable is encoded with -1,0 ,+1 respectively, and model is passed through minimum
Two, which multiply solution, is fitted secondary multinomial equation.
4 response surface experimental design factor level table of table
5 response surface experimental program of table
Tested number | X1 | X2 | X3 |
1 | -1 | -1 | 0 |
2 | 1 | -1 | 0 |
3 | -1 | 1 | 0 |
4 | 1 | 1 | 0 |
5 | -1 | 0 | -1 |
6 | 1 | 0 | -1 |
7 | -1 | 0 | 1 |
8 | 1 | 0 | 1 |
9 | 0 | -1 | -1 |
10 | 0 | 1 | -1 |
11 | 0 | -1 | 1 |
12 | 0 | 1 | 1 |
13 | 0 | 0 | 0 |
14 | 0 | 0 | 0 |
15 | 0 | 0 | 0 |
16 | 0 | 0 | 0 |
17 | 0 | 0 | 0 |
It is tested according to table 4 and 5, laboratory sample carries out the measurement of protein content, and result is inserted in following table.
6 response surface contrived experiment result of table
Experimental result significance analysis such as table 7.
The results of analysis of variance of 7 regression equation of table
Data source | Quadratic sum SS | Side MS | F | Pr>F |
Model | 1.86E+0.05 | 20651.89 | 8.31 | 0.0054 |
X1 | 23051.91 | 23051.91 | 9.27 | 0.0187 |
X2 | 37646.45 | 37646.45 | 15.15 | 0.0060 |
X3 | 972.07 | 972.07 | 0.39 | 0.5516 |
X1 2 | 34415.95 | 34415.95 | 13.85 | 0.0074 |
X2 2 | 6652.98 | 6652.98 | 2.68 | 0.15 |
X3 2 | 36766.38 | 36766.38 | 14.79 | 0.0063 |
X1X2 | 23135.47 | 23135.47 | 9.31 | 0.018 |
X1X3 | 7270.03 | 7270.03 | 2.92 | 0.13 |
X2X3 | 8083.09 | 8083.09 | 3.25 | 0.11 |
Residual error | 17398.94 | 2485.56 | ||
It is quasi- poor to lose | 1632.55 | 5443.18 | 20.36 | 0.069 |
Pure error | 1069.38 | 267.35 | ||
Total deviation | 2.03E+0.05 |
It is fitted the foundation of regression equation:Fitting of a polynomial recurrence is carried out to experimental data, total nitrogen content (Y) is dependent variable,
Ultrasonic time (X1), ultrasonic temperature (X2), extraction solid-liquid ratio (X3) are independent variable, and it is as follows to establish regression equation:
Y=89.52-3.06X1+15.51X2-1737.45X3-0.11X1X2-43.82X1X3+17.3 7X2X3+
0.12X12-0.16X22+10479.82X32
It using total nitrogen content as index, is analyzed by Design-Expert8.0.6 softwares, obtained optimum results are:
The ultrasonic extraction time is 30minutes, ultrasonic extraction temperature is 49.42 DEG C, solid-liquid ratio 1:5g/ml, due to practical operation
Control problem, the temperature of ultrasonic extraction, which takes 50 DEG C, to be advisable.
When being extracted with optimal conditions, the protein content that prediction extraction measures will be 307.28mg/g, to extract institute
The highest protein content obtained.Consider practical operation, carries out actual extracting, recovery rate 328.32mg/g, with reality with this condition
It tests predicted value to be substantially consistent, it was confirmed that the accuracy and practicability of contrived experiment.
Experiment 5:
Sandworm extract is climbed using what is be prepared in embodiment 1, test climbs sandworm extract to hypoimmunity mouse
The protecting effect of specific humoral immunity:
KM kinds mouse 24, half male and half female, adaptability raise 7d, and room temperature control is at 25 DEG C, relative humidity 78-81%, 12h
Illumination that day alternates with night, ad lib and drinking-water.Experiment before each group mouse be deprived of food but not water 12h, weigh, according to mouse weight with
Machine is divided into 4 groups, respectively:Normal group, cyclophosphamide model group climb sandworm high dose group and climb sandworm low dose group, often
Group 6.The daily gavage of each group mouse 1 time, continuous 11d, Normal group and cyclophosphamide model group gavage (ig) are steamed in equal volume
Distilled water, it is crude drug amount 2g.kg to climb sandworm high dose group ig dosage-1.d-1, low dose group ig dosage be 1g.kg-1.d-1, gavage body
Product 10ml.kg-1.d-1.In gavage the 5th, 7,9,11d, (ip.) distilled water 10ml.kg is injected intraperitoneally in Normal group-1, ring phosphorus
Amide model group climbs sandworm high dose group and climbs sandworm low dose group difference ip. cyclophosphamide 80mg.kg-1, it is low to prepare immunity
Under mouse model.After normally 2h is administered in 11d, mouse plucks eyeball and takes blood, 37 DEG C of incubation 30min, 4000r/min centrifugations
5min takes supernatant both to obtain serum, is operated according to IgM kit specifications, measures mice serum IgM contents.
Mice serum IgM assays the results are shown in Table 8, as shown in Table 8:Compared with Normal group, cyclophosphamide model
Group significantly reduces the concentration (P of IgM in the serum of mouse<0.05), explanation establishes the low mould of mouse immunity with cyclophosphamide
Type success.Compared with cyclophosphamide model group, climbs sandworm low dose group and climb sandworm high dose group serum IgM level and obviously rise
Height (P<0.05).Showing to climb sandworm water-soluble extractive (1000,2000mg/kg) has IgM in hypoimmunity mice serum
It is obviously improved.Sandworm water-soluble extractive is climbed in prompt has apparent protection to the specific humoral immunity of hypoimmunity mouse
Effect.
Table 8 climbs influence of the sandworm water-soluble extractive to mice serum IgM levels
# expressions are compared with normal group, #P < 0.05;* it indicates compared with model group, * P < 0.05.
Experiment 6:
Sandworm extract is climbed using what is be prepared in embodiment 1, test climbs sandworm extract to hypoimmunity mouse
The protective effect of cellular immunity.
KM kinds mouse 24, half male and half female, grouping and administering mode are the same as example five.But mouse peritoneal is noted in gavage 6d
20% rabbit erythrocyte is penetrated, 0.2ml/ is only.After 11d is normally administered 2 hours, serum is made with five method of example, using the side ELISA
Method measures mice serum IL-2 contents.
Mice serum IL-2 assays the results are shown in Table 9, as shown in Table 9, compared with Normal group, cyclophosphamide model
Group significantly reduces the concentration (P of the red immunized mice serum IL-2 of rabbit<0.05), explanation establishes mouse immunity with cyclophosphamide
Low model success.Compared with cyclophosphamide model group, it is horizontal apparent to climb sandworm water-soluble extractive high dose group serum IL-2
Increase (P<0.05).Show to climb sandworm water-soluble extractive (2000mg/kg) have to IL-2 in hypoimmunity mice serum it is bright
Aobvious promotion.Sandworm water-soluble extractive is climbed in prompt has significant protective effect to the cellular immunity of hypoimmunity mouse.
Table 9 climbs influence of the sandworm water-soluble extractive to mice serum IL-2 levels
# expressions are compared with normal group, #P < 0.05;* it indicates compared with model group, * P < 0.05.
Influence of the sandworm extract to hypoimmunity mouse organs' index is climbed in experiment 7
According to experiment five, after each group mouse is intervened with relative medicine, after the completion of blood sampling, mouse is taken off into neck and is put to death, dissection is plucked
Spleen and thymus gland are taken, mice spleen quality and thymic factor D injection are accurately weighed with electronic balance, organ index is calculated as follows.
Organ index=Organ weight (mg)/mouse weight (g) x 100%
It climbs influence of the sandworm extract to caused by cyclophosphamide hypoimmunity mouse thymus index and spleen index and is shown in Table 10,
As shown in Table 10, compared with Normal group, cyclophosphamide group significantly reduces the thymus index and spleen index (P of mouse<
0.01), explanation establishes the low model success of mouse immunity with cyclophosphamide;Compared with cyclophosphamide model group, sandworm water is climbed
Soluble extract high dose group thymus index apparent increase (P<0.05);It climbs sandworm low dose group and high dose group spleen index is apparent
Increase (P<0.01).Climbing sandworm water-soluble extractive makes the thymus gland of hypoimmunity mouse and spleen quality increase, and prompts it can be with
Stimulation thymus gland and spleen generate immunocyte and enhance the immune function of body.
Table 10 climbs influence of the sandworm water-soluble extractive to mouse organs' index
# expressions are compared with normal group, ##P < 0.01;* it indicates compared with model group, * P < 0.05.
The above case study on implementation is merely to illustrate the technical solution of this patent, rather than its limitations;Although with reference to aforementioned implementation
This patent is described in detail in case, it will be understood by those of ordinary skill in the art that:It still can be to aforementioned each
Technical solution recorded in case study on implementation is modified or equivalent replacement of some of the technical features;And these are repaiied
Change or replaces, the spirit and scope for each case study on implementation technical solution of this patent that it does not separate the essence of the corresponding technical solution.
Example described above is only the preferred embodiment of this patent, but the protection domain of this patent is not limited thereto.
It should be pointed out that for those skilled in the art, under the premise of not departing from this patent principle, specially according to this
The technical solution and its inventional idea of profit, can also make several improvements and modifications, these improvements and modifications also should be regarded as this specially
The protection domain of profit.
Claims (4)
1. a kind of preparation method for climbing sandworm extract adjusting immune function, it is characterised in that include the following steps:
1)Climb the preparation of sandworm coarse powder:The fresh and alive sandworm that climbs is cleaned with pure water, is removed end to end and internal organ, is cleaned and drain, in-
80 DEG C of quick freezes are spare;
2)Before active constituent extraction, done in freezing with blender by its broken fritter for 5mm with 25 DEG C of warm water constant-temperature thawings
It is freeze-dried 36h in dry machine;Dry polypide was broken into 40 mesh sieve with pulverizer, it is spare at a temperature of being sealed in -20 DEG C;
3)Climb the extraction of sandworm extract:By solid-liquid ratio 1:2-1:Distilled water is added in 4 ratio, impregnates half using 25 DEG C of warm water
A hour is extracted with ultrasonic wave drug processor, and filtering takes filtrate, and upper filtrate, which is placed in freeze drier, to be dried 36 hours, is received
Collect the sample after drying to get sandworm extract is climbed.
2. adjusting the preparation method for climbing sandworm extract of immune function as described in claim 1, it is characterised in that:Be set forth in-
Spare 80 DEG C of quick freezes refer to needing its adfreezing in 20 minutes.
3. a kind of further preparing the method for climbing sandworm skimmed milk, spy according to method described in claim 1 or claim 2
Sign is:Step 3 '), by step 2)Last spare substance is according to solid-liquid ratio 1:5-1:Petroleum ether is added in 10 ratio, in 20-
After 40 DEG C of water-bath reflux 2h, petroleum ether is recovered by filtration, obtains climbing sandworm skimmed milk.
4. a kind of further preparing the technique for climbing sandworm water extract coarse powder according to method described in claim 3, feature exists
It is in its step:It takes and climbs sandworm skimmed milk, be 1 according to solid-liquid ratio:4-1:Pure water is added in 20 ratio, is assisted using ultrasonic wave
Filtrate, then the miillpore filter mistake with 0.45nm is collected by filtration in extraction method, 15-60 DEG C of Extracting temperature, extraction time 15-60min
Filtrate is collected in filter, is freeze-dried in freeze drier to get sandworm water extract coarse powder is climbed.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109498659A (en) * | 2018-12-22 | 2019-03-22 | 广东医科大学 | Application of the sandworm zymolyte in preparation prevention and treatment osteoarthritis drugs or health care product |
CN109938342A (en) * | 2019-04-01 | 2019-06-28 | 四川轻化工大学 | Preparation method of corydalus cornutus extract with effect of correcting kidney-yang deficiency |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1559447A (en) * | 2004-02-18 | 2005-01-05 | 中国人民解放军军需大学 | Extracte of Aspongopus chinensis Dallas and Extraction method therefor medicinal use and preparn. thereof |
CN101991608A (en) * | 2010-11-19 | 2011-03-30 | 浙江省医学科学院 | Fly maggot extractive as well as preparation method and application thereof |
CN103948635A (en) * | 2014-04-30 | 2014-07-30 | 四川锡成天然食品有限公司 | Method for preparing freeze-dried larvae of pasha worms |
CN104531333A (en) * | 2014-12-12 | 2015-04-22 | 四川农业大学 | Acquiring method of novel oil |
-
2018
- 2018-06-08 CN CN201810586283.0A patent/CN108741056A/en active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1559447A (en) * | 2004-02-18 | 2005-01-05 | 中国人民解放军军需大学 | Extracte of Aspongopus chinensis Dallas and Extraction method therefor medicinal use and preparn. thereof |
CN101991608A (en) * | 2010-11-19 | 2011-03-30 | 浙江省医学科学院 | Fly maggot extractive as well as preparation method and application thereof |
CN103948635A (en) * | 2014-04-30 | 2014-07-30 | 四川锡成天然食品有限公司 | Method for preparing freeze-dried larvae of pasha worms |
CN104531333A (en) * | 2014-12-12 | 2015-04-22 | 四川农业大学 | Acquiring method of novel oil |
Non-Patent Citations (4)
Title |
---|
李军德,等: "《中国药用动物志 上 第2版》", 30 June 2013 * |
李勇,等: "东方巨齿蛉幼虫的营养成分测定", 《贵州农业科学》 * |
李奕冉,等: "昆虫蛋白对动物免疫功能影响研究进展", 《动物医学进展》 * |
杨冬梅,等: "爬沙虫提取液对果蝇繁殖力及寿命的影响", 《天然产物研究与开发》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109498659A (en) * | 2018-12-22 | 2019-03-22 | 广东医科大学 | Application of the sandworm zymolyte in preparation prevention and treatment osteoarthritis drugs or health care product |
CN109498659B (en) * | 2018-12-22 | 2022-03-15 | 广东医科大学 | Application of sandworm zymolyte in preparation of osteoarthritis prevention and treatment medicines or health-care products |
CN109938342A (en) * | 2019-04-01 | 2019-06-28 | 四川轻化工大学 | Preparation method of corydalus cornutus extract with effect of correcting kidney-yang deficiency |
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