CN106416992B - A kind of Enteromorpha special nutrient fluid and preparation method thereof - Google Patents
A kind of Enteromorpha special nutrient fluid and preparation method thereof Download PDFInfo
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- CN106416992B CN106416992B CN201610858947.5A CN201610858947A CN106416992B CN 106416992 B CN106416992 B CN 106416992B CN 201610858947 A CN201610858947 A CN 201610858947A CN 106416992 B CN106416992 B CN 106416992B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G33/00—Cultivation of seaweed or algae
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
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Abstract
The present invention relates to a kind of Enteromorpha special nutrient fluids, further relate to a kind of preparation method of above-mentioned Enteromorpha special nutrient fluid, the preparation method is the following steps are included: (1) sterilizes seawater；(2) nutrient is added into seawater and is mixed；(3) antipathogenic composition is weighed in proportion；(4) pennywort in antipathogenic composition, cactus, onion are smashed to pieces or is crushed and take gained juice；(5) cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic are decocted and takes extracting solution；(6) Cortex hibisus syriacus, the achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice root are ground, obtains grounds travel；(7) effective component in grounds travel is extracted, and is concentrated to give medicinal extract；(8) a certain proportion of medicinal extract is added into seawater, mixes.It is an advantage of the invention that sterilization culture effect is good, and be conducive to the growth of Enteromorpha.
Description
Technical field
The invention belongs to algae culture technical field more particularly to a kind of Enteromorpha special nutrient fluids, further relate to a kind of above-mentioned
The preparation method of Enteromorpha special nutrient fluid.
Background technique
Enteromorpha is a category of Chlorophyta Ulvaceae, and there are about 40 kinds, there are about 11 kinds for China.Chinese frequent species are had chance with pipe waterside
Tongue fur, flat Enteromorpha, Enteromorpha, most type marine products are widely distributed in each ocean in the whole world, and some types are in brackish water or rivers
In also show.Enteromorpha is frequently grown on the rock of intertidal zone, on the chad on silt beach, can also be grown nonparasitically upon another plant sometimes in the algae of tangleweed
On body.Due to Global climate change, water eutrophication etc., ocean tangleweed Enteromorpha green tide is caused to break out.Though Enteromorpha
It is so nontoxic, but as red tide, prolific Enteromorpha also can shielded from sunlight, influence the growth of seabed algae；Dead waterside
Tongue fur can also consume the oxygen in seawater；There are also studies have shown that the chemical substance of Enteromorpha secretion is likely to also give birth to other oceans
Object adversely affects.In addition, Enteromorpha outburst can also seriously affect landscape, the progress gone sightseeing with aquatic sports is interfered.
Enteromorpha be frequently grown on the rock of intertidal zone or Shi Zhaozhong or the chad on silt beach on, can also grow nonparasitically upon another plant in large size sometimes
On the frond of seaweed.Under such growing environment, Enteromorpha inevitably with many miscellaneous algaes and miscellaneous bacteria there is symbiosis, because
This, the Enteromorpha algae sample adopted back from field is also unable to reach aseptic result even if cultivating a very long time in the lab.But
It is, for interaction among algae and for the microexamination of frond, to require with no bacterium algae to be real in the research for Enteromorpha
Material is tested, just can ensure that therefore the reliability of Enteromorpha research obtains sterile algae and Enteromorpha in laboratory is studied to Guan Chong
It wants.Currently, the sterilization culture of Enteromorpha mainly uses sterilization, physical method, antibiotic method etc., however the effect of its sterilization culture
Fruit is bad, and can influence the growth of Enteromorpha to a certain extent.
Summary of the invention
An object of the present invention is to provide a kind of preparation method of Enteromorpha special nutrient fluid, is the growth according to Enteromorpha
Feature preparation, sterilization culture effect is good, and is conducive to the growth of Enteromorpha.
In order to solve the above-mentioned technical problem, the invention adopts the following technical scheme: a kind of preparation of Enteromorpha special nutrient fluid
Method, comprising the following steps:
(1) stand-by seawater is put into high-pressure steam sterilizing pan, is sterilized 20-30 minutes under the conditions of 121 DEG C；
(2) 0.04-0.05g NaNO is added in the seawater after every 1L sterilizing3、0.01g Na2HPO4·12H2O、0.02mg
MnCL2·4H2O、0.25-0.3mg FeSO4·7H2O、3.5-4mgNa20.2mgVB is added in EDTA after mixing1、
0.001mgVB12, 0.001mg biotin, be uniformly mixed；
(3) weigh each component of following mass fraction: 32-37 parts of pennywort, 30-35 parts of cactus, 30-35 parts of onion,
27-33 parts of cordate houttuynia, 21-28 parts of frutus cnidii, 21-26 parts of lophatherum gracile, 20-27 parts of dandelion, 18-25 parts of Prunella vulgaris, kiss-me
17-21 parts, 9-15 parts of peppermint oil, 14-20 parts of garlic, 12-18 parts of Cortex hibisus syriacus, 11-17 parts of the achene of Siberian cocklebur, 10-17 parts of kaempferia galanga, great Wu
Steep root 8-15 parts, 7-15 parts of Indian Epimeredi Herb, 6-10 parts of lotus nut, 6-10 parts of mulberry leaf, 5-9 parts of phellodendron bark, 5-9 parts of glutinous rice root；
(4) load weighted pennywort, cactus, onion are smashed to pieces or is crushed and take gained juice；
(5) load weighted cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic are crushed respectively,
Addition quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 15 times of clear water,
After 20-30 DEG C of immersion 5-6h, it is heated to the 1h that boils and be cooked by slow fire, fried liquid is taken out in separation；Continue to be added into drug residue
Quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 10 times of clear water, be heated to
It boils and is cooked by slow fire 1h, merging fried liquid twice is extracting solution；
(6) by load weighted Cortex hibisus syriacus, the achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice
Root is packed into ball mill and is fully ground, and crosses 80 meshes, obtains grounds travel；
(7) extracting solution and step (6) obtained load weighted peppermint oil, the resulting juice of step (4) and step (5)
Obtained grounds travel is placed in the extraction kettle in supercritical extract instrument, is utilized carbon dioxide as medium and is extracted, extraction
Kettle pressure is 40Mpa, and extraction temperature is 45 DEG C, separator pressure 12Mpa, and separator temperature is 70 DEG C, extracts 2 hours, obtains
Extract liquor；Extract liquor is concentrated in outer circulation inspissator, condition is concentrated are as follows: pressure 0.03Mpa, temperature 70 C are concentrated into phase
The medicinal extract for being 1.12~1.14 to density；
(8) 1-1.2g step (7) resulting medicinal extract is added into every liter of resulting seawater of step (2), and is uniformly mixed, i.e.,
Obtain object.
Enteromorpha special nutrient fluid of the invention is according to Enteromorpha growth characteristic development, and nutritive value is high, contains Enteromorpha
Microelement necessary to growing and nutrient, and nutrition is easy to be absorbed by Enteromorpha, conducive to the growth of Enteromorpha.Step (1)
Operation the purpose of be kill seawater in bacterium.The component of step (2) and step (3) is with above-mentioned special ratios and preparation method system
Standby resulting Enteromorpha nutrient solution is full of nutrition, is able to satisfy Enteromorpha growth needs, promotes Enteromorpha fast-growth, and can inhibit other
Bacterium or seaweed growth successfully obtain sterile Enteromorpha, are conducive to the subsequent laboratory research of Enteromorpha.Step will weigh in (4)
Pennywort, cactus, the onion purpose smashing to pieces or crush be conducive to pennywort, cactus, the antipathogenic composition in onion and
Nutritional ingredient preferably plays a role in subsequent extraction step.In step (5), by load weighted cordate houttuynia, frutus cnidii, henon bamboo
Leaf, dandelion, Prunella vulgaris, kiss-me, garlic crush effect be conducive to impregnate and decoction process in each component it is effective at
Point be more fully precipitated, be with 20-30 DEG C of clear water immersions 5-6h in order to make the effective component of each component be easier precipitation, decoction
Effect is the irritation effect that can change drug, relatively mild to act on Enteromorpha.In step (6), by Cortex hibisus syriacus, the achene of Siberian cocklebur,
Kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice root are fitted into ball mill and are fully ground and can make in medicinal material
Effective component is more fully precipitated in subsequent step.In step (7), supercritical extract step can retain this hair to greatest extent
The effective component of bright middle each component.The Enteromorpha of field acquisition is subjected to sterilization training with nutrient solution of the invention in the lab
Support, experimental study shows that the Enteromorpha speed of growth is fast in sterilization incubation, Enteromorpha green in color, by sterilization culture at
Ripe frond is laid in LB solid culture primary surface, cultivates 10 days, does not there is varied bacteria growing.
Preferably, in step (2), 0.042g NaNO is added in the culture seawater after every 1L sterilizing3、0.01gNa2HPO4·
12H2O、0.02mg MnCL2·4H2O、0.28mg FeSO4·7H2O、3.7mgNa2EDTA is uniformly mixed, is then added
0.2mgVB1、0.001mgVB12, 0.001mg biotin, be uniformly mixed.
Preferably, in step (2), 0.05g NaNO is added in the culture seawater after every 1L sterilizing3、0.01gNa2HPO4·
12H2O、0.02mg MnCL2·4H2O、0.25mg FeSO4·7H2O、3.8mgNa2EDTA is uniformly mixed, is then added
0.2mgVB1、0.001mgVB12, 0.001mg biotin, be uniformly mixed.
Preferably, in step (2), 0.043g NaNO is added in the culture seawater after every 1L sterilizing3、0.01gNa2HPO4·
12H2O、0.02mg MnCL2·4H2O、0.3mg FeSO4·7H2O、4mgNa2EDTA is uniformly mixed, is then added
0.2mgVB1、0.001mgVB12, 0.001mg biotin, be uniformly mixed.
Preferably, in step (3), each component of following mass fraction: 32 parts of pennywort, 33 parts of cactus, onion is weighed
30 parts, 33 parts of cordate houttuynia, 28 parts of frutus cnidii, 25 parts of lophatherum gracile, 27 parts of dandelion, 18 parts of Prunella vulgaris, 17 parts of kiss-me, peppermint oil
9 parts, 15 parts of garlic, 12 parts of Cortex hibisus syriacus, 17 parts of the achene of Siberian cocklebur, 16 parts of kaempferia galanga, 15 parts of szechwan raspberry root or leaf, 14 parts of Indian Epimeredi Herb, 10 parts of lotus nut,
10 parts of mulberry leaf, 9 parts of phellodendron bark, 8 parts of glutinous rice root.
Preferably, in step (3), each component of following mass fraction: 34 parts of pennywort, 30 parts of cactus, onion is weighed
33 parts, 31 parts of cordate houttuynia, 25 parts of frutus cnidii, 26 parts of lophatherum gracile, 25 parts of dandelion, 25 parts of Prunella vulgaris, 21 parts of kiss-me, peppermint oil
12 parts, 14 parts of garlic, 15 parts of Cortex hibisus syriacus, 11 parts of the achene of Siberian cocklebur, 14 parts of kaempferia galanga, 8 parts of szechwan raspberry root or leaf, 15 parts of Indian Epimeredi Herb, 9 parts of lotus nut,
9 parts of mulberry leaf, 8 parts of phellodendron bark, 9 parts of glutinous rice root.
Preferably, in step (3), each component of following mass fraction: 35 parts of pennywort, 35 parts of cactus, onion is weighed
32 parts, 27 parts of cordate houttuynia, 23 parts of frutus cnidii, 22 parts of lophatherum gracile, 20 parts of dandelion, 20 parts of Prunella vulgaris, 18 parts of kiss-me, peppermint oil
15 parts, 17 parts of garlic, 18 parts of Cortex hibisus syriacus, 12 parts of the achene of Siberian cocklebur, 17 parts of kaempferia galanga, 10 parts of szechwan raspberry root or leaf, 7 parts of Indian Epimeredi Herb, 6 parts of lotus nut,
6 parts of mulberry leaf, 5 parts of phellodendron bark, 7 parts of glutinous rice root.
Preferably, in step (3), each component of following mass fraction: 37 parts of pennywort, 32 parts of cactus, onion is weighed
35 parts, 29 parts of cordate houttuynia, 21 parts of frutus cnidii, 21 parts of lophatherum gracile, 22 parts of dandelion, 21 parts of Prunella vulgaris, 20 parts of kiss-me, peppermint oil
11 parts, 20 parts of garlic, 16 parts of Cortex hibisus syriacus, 15 parts of the achene of Siberian cocklebur, 10 parts of kaempferia galanga, 12 parts of szechwan raspberry root or leaf, 10 parts of Indian Epimeredi Herb, 8 parts of lotus nut,
8 parts of mulberry leaf, 7 parts of phellodendron bark, 5 parts of glutinous rice root.
The invention also discloses Enteromorpha special nutrient fluids obtained by a kind of above-mentioned preparation method.
Compared with prior art, the beneficial effects of the present invention are: Enteromorpha special nutrient fluid of the invention is according to Enteromorpha
Growth characteristic development, nutritive value is high, microelement necessary to growing containing Enteromorpha and nutrient, and nutrition is held
It is easily absorbed by Enteromorpha, conducive to the growth of Enteromorpha.The purpose of operation of step (1) is the bacterium killed in seawater.Step (2) and step
(3) it is full of nutrition that component with above-mentioned special ratios and preparation method prepares resulting Enteromorpha nutrient solution, is able to satisfy Enteromorpha growth
It needs, promotes Enteromorpha fast-growth, and other bacteriums or seaweed growth can be inhibited, successfully obtain sterile Enteromorpha, be conducive to waterside
The subsequent laboratory research of tongue fur.It by the purpose that load weighted pennywort, cactus, onion are smashed to pieces or crush is to have in step (4)
Conducive in pennywort, cactus, onion antipathogenic composition and nutritional ingredient preferably play a role in subsequent extraction step.Step
Suddenly in (5), it is by the effect that load weighted cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic crush
Be conducive to impregnate and decoction process in the effective component of each component be more fully precipitated, with 20-30 DEG C of immersion 5-6h of clear water be for
The effective component of each component is set to be easier to be precipitated, the effect of decoction is the irritation effect that can change drug, relatively mild
Act on Enteromorpha.In step (6), by the Cortex hibisus syriacus, achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous
Rice root, which is fitted into ball mill and is fully ground, can be such that the effective component in medicinal material is more fully precipitated in subsequent step.In step (7),
Supercritical extract step can retain the effective component of each component in the present invention to greatest extent.Originally by the Enteromorpha of field acquisition
The nutrient solution of invention carries out sterilization culture in the lab, and experimental study shows Enteromorpha growth speed in sterilization incubation
Degree is fast, Enteromorpha green in color, and the mature frond of sterilization culture is laid in LB solid culture primary surface, cultivates 10 days, does not have
Varied bacteria growing.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.
Embodiment 1
A kind of preparation method of Enteromorpha special nutrient fluid, comprising the following steps:
(1) stand-by seawater is put into high-pressure steam sterilizing pan, is sterilized 20-30 minutes under the conditions of 121 DEG C；
(2) 0.04g NaNO is added in the seawater after every 1L sterilizing3、0.01g Na2HPO4·12H2O、0.02mgMnCL2·
4H2O、0.26mg FeSO4·7H2O、3.5mgNa20.2mgVB is added in EDTA after mixing1、0.001mgVB12、0.001mg
Biotin is uniformly mixed；
(3) each component of following mass fraction: 37 parts of pennywort, 32 parts of cactus, 35 parts of onion, cordate houttuynia 29 is weighed
Part, 21 parts of frutus cnidii, 21 parts of lophatherum gracile, 22 parts of dandelion, 21 parts of Prunella vulgaris, 20 parts of kiss-me, 11 parts of peppermint oil, garlic 20
Part, 16 parts of Cortex hibisus syriacus, 15 parts of the achene of Siberian cocklebur, 10 parts of kaempferia galanga, 12 parts of szechwan raspberry root or leaf, 10 parts of Indian Epimeredi Herb, 8 parts of lotus nut, 8 parts of mulberry leaf, Huang
7 parts of cypress skin, 5 parts of glutinous rice root；
(4) load weighted pennywort, cactus, onion are smashed to pieces or is crushed and take gained juice；
(5) load weighted cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic are crushed respectively,
Addition quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 15 times of clear water,
After 20-30 DEG C of immersion 5-6h, it is heated to the 1h that boils and be cooked by slow fire, fried liquid is taken out in separation；Continue to be added into drug residue
Quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 10 times of clear water, be heated to
It boils and is cooked by slow fire 1h, merging fried liquid twice is extracting solution；
(6) by load weighted Cortex hibisus syriacus, the achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice
Root is packed into ball mill and is fully ground, and crosses 80 meshes, obtains grounds travel；
(7) extracting solution and step (6) obtained load weighted peppermint oil, the resulting juice of step (4) and step (5)
Obtained grounds travel is placed in the extraction kettle in supercritical extract instrument, is utilized carbon dioxide as medium and is extracted, extraction
Kettle pressure is 40Mpa, and extraction temperature is 45 DEG C, separator pressure 12Mpa, and separator temperature is 70 DEG C, extracts 2 hours, obtains
Extract liquor；Extract liquor is concentrated in outer circulation inspissator, condition is concentrated are as follows: pressure 0.03Mpa, temperature 70 C are concentrated into phase
The medicinal extract for being 1.12~1.14 to density；
(8) 1-1.2g step (7) resulting medicinal extract is added into every liter of resulting seawater of step (2), and is uniformly mixed, i.e.,
Obtain object.
Enteromorpha special nutrient fluid obtained by a kind of above-mentioned preparation method.
Embodiment 2
A kind of preparation method of Enteromorpha special nutrient fluid, comprising the following steps:
(1) stand-by seawater is put into high-pressure steam sterilizing pan, is sterilized 20-30 minutes under the conditions of 121 DEG C；
(2) 0.042g NaNO is added in the seawater after every 1L sterilizing3、0.01gNa2HPO4·12H2O、0.02mgMnCL2·
4H2O、0.28mg FeSO4·7H2O、3.7mgNa2EDTA is uniformly mixed, 0.2mgVB is then added1、0.001mgVB12、
0.001mg biotin is uniformly mixed；
(3) each component of following mass fraction: 32 parts of pennywort, 33 parts of cactus, 30 parts of onion, cordate houttuynia 33 is weighed
Part, 28 parts of frutus cnidii, 25 parts of lophatherum gracile, 27 parts of dandelion, 18 parts of Prunella vulgaris, 17 parts of kiss-me, 9 parts of peppermint oil, 15 parts of garlic,
12 parts of Cortex hibisus syriacus, 17 parts of the achene of Siberian cocklebur, 16 parts of kaempferia galanga, 15 parts of szechwan raspberry root or leaf, 14 parts of Indian Epimeredi Herb, 10 parts of lotus nut, 10 parts of mulberry leaf, Cortex Phellodendri
9 parts of skin, 8 parts of glutinous rice root；
(4) load weighted pennywort, cactus, onion are smashed to pieces or is crushed and take gained juice；
(5) load weighted cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic are crushed respectively,
Addition quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 15 times of clear water,
After 20-30 DEG C of immersion 5-6h, it is heated to the 1h that boils and be cooked by slow fire, fried liquid is taken out in separation；Continue to be added into drug residue
Quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 10 times of clear water, be heated to
It boils and is cooked by slow fire 1h, merging fried liquid twice is extracting solution；
(6) by load weighted Cortex hibisus syriacus, the achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice
Root is packed into ball mill and is fully ground, and crosses 80 meshes, obtains grounds travel；
(7) extracting solution and step (6) obtained load weighted peppermint oil, the resulting juice of step (4) and step (5)
Obtained grounds travel is placed in the extraction kettle in supercritical extract instrument, is utilized carbon dioxide as medium and is extracted, extraction
Kettle pressure is 40Mpa, and extraction temperature is 45 DEG C, separator pressure 12Mpa, and separator temperature is 70 DEG C, extracts 2 hours, obtains
Extract liquor；Extract liquor is concentrated in outer circulation inspissator, condition is concentrated are as follows: pressure 0.03Mpa, temperature 70 C are concentrated into phase
The medicinal extract for being 1.12~1.14 to density；
(8) 1-1.2g step (7) resulting medicinal extract is added into every liter of resulting seawater of step (2), and is uniformly mixed, i.e.,
Obtain object.
Enteromorpha special nutrient fluid obtained by a kind of above-mentioned preparation method.
Embodiment 3
A kind of preparation method of Enteromorpha special nutrient fluid, comprising the following steps:
(1) stand-by seawater is put into high-pressure steam sterilizing pan, is sterilized 20-30 minutes under the conditions of 121 DEG C；
(2) 0.05g NaNO is added in the seawater after every 1L sterilizing3、0.01gNa2HPO4·12H2O、0.02mgMnCL2·
4H2O、0.25mg FeSO4·7H2O、3.8mgNa2EDTA is uniformly mixed, 0.2mgVB is then added1、0.001mgVB12、
0.001mg biotin is uniformly mixed；
(3) each component of following mass fraction: 34 parts of pennywort, 30 parts of cactus, 33 parts of onion, cordate houttuynia 31 is weighed
Part, 25 parts of frutus cnidii, 26 parts of lophatherum gracile, 25 parts of dandelion, 25 parts of Prunella vulgaris, 21 parts of kiss-me, 12 parts of peppermint oil, garlic 14
Part, 15 parts of Cortex hibisus syriacus, 11 parts of the achene of Siberian cocklebur, 14 parts of kaempferia galanga, 8 parts of szechwan raspberry root or leaf, 15 parts of Indian Epimeredi Herb, 9 parts of lotus nut, 9 parts of mulberry leaf, Cortex Phellodendri
8 parts of skin, 9 parts of glutinous rice root；
(4) load weighted pennywort, cactus, onion are smashed to pieces or is crushed and take gained juice；
(5) load weighted cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic are crushed respectively,
Addition quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 15 times of clear water,
After 20-30 DEG C of immersion 5-6h, it is heated to the 1h that boils and be cooked by slow fire, fried liquid is taken out in separation；Continue to be added into drug residue
Quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 10 times of clear water, be heated to
It boils and is cooked by slow fire 1h, merging fried liquid twice is extracting solution；
(6) by load weighted Cortex hibisus syriacus, the achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice
Root is packed into ball mill and is fully ground, and crosses 80 meshes, obtains grounds travel；
(7) extracting solution and step (6) obtained load weighted peppermint oil, the resulting juice of step (4) and step (5)
Obtained grounds travel is placed in the extraction kettle in supercritical extract instrument, is utilized carbon dioxide as medium and is extracted, extraction
Kettle pressure is 40Mpa, and extraction temperature is 45 DEG C, separator pressure 12Mpa, and separator temperature is 70 DEG C, extracts 2 hours, obtains
Extract liquor；Extract liquor is concentrated in outer circulation inspissator, condition is concentrated are as follows: pressure 0.03Mpa, temperature 70 C are concentrated into phase
The medicinal extract for being 1.12~1.14 to density；
(8) 1-1.2g step (7) resulting medicinal extract is added into every liter of resulting seawater of step (2), and is uniformly mixed, i.e.,
Obtain object.
Enteromorpha special nutrient fluid obtained by a kind of above-mentioned preparation method.
Embodiment 4
A kind of preparation method of Enteromorpha special nutrient fluid, comprising the following steps:
(1) stand-by seawater is put into high-pressure steam sterilizing pan, is sterilized 20-30 minutes under the conditions of 121 DEG C；
(2) 0.043g NaNO is added in the seawater after every 1L sterilizing3、0.01gNa2HPO4·12H2O、0.02mgMnCL2·
4H2O、0.3mg FeSO4·7H2O、4mgNa2EDTA is uniformly mixed, 0.2mgVB is then added1、0.001mgVB12、0.001mg
Biotin is uniformly mixed；
(3) each component of following mass fraction: 35 parts of pennywort, 35 parts of cactus, 32 parts of onion, cordate houttuynia 27 is weighed
Part, 23 parts of frutus cnidii, 22 parts of lophatherum gracile, 20 parts of dandelion, 20 parts of Prunella vulgaris, 18 parts of kiss-me, 15 parts of peppermint oil, garlic 17
Part, 18 parts of Cortex hibisus syriacus, 12 parts of the achene of Siberian cocklebur, 17 parts of kaempferia galanga, 10 parts of szechwan raspberry root or leaf, 7 parts of Indian Epimeredi Herb, 6 parts of lotus nut, 6 parts of mulberry leaf, Cortex Phellodendri
5 parts of skin, 7 parts of glutinous rice root；
(4) load weighted pennywort, cactus, onion are smashed to pieces or is crushed and take gained juice；
(5) load weighted cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic are crushed respectively,
Addition quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 15 times of clear water,
After 20-30 DEG C of immersion 5-6h, it is heated to the 1h that boils and be cooked by slow fire, fried liquid is taken out in separation；Continue to be added into drug residue
Quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 10 times of clear water, be heated to
It boils and is cooked by slow fire 1h, merging fried liquid twice is extracting solution；
(6) by load weighted Cortex hibisus syriacus, the achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice
Root is packed into ball mill and is fully ground, and crosses 80 meshes, obtains grounds travel；
(7) extracting solution and step (6) obtained load weighted peppermint oil, the resulting juice of step (4) and step (5)
Obtained grounds travel is placed in the extraction kettle in supercritical extract instrument, is utilized carbon dioxide as medium and is extracted, extraction
Kettle pressure is 40Mpa, and extraction temperature is 45 DEG C, separator pressure 12Mpa, and separator temperature is 70 DEG C, extracts 2 hours, obtains
Extract liquor；Extract liquor is concentrated in outer circulation inspissator, condition is concentrated are as follows: pressure 0.03Mpa, temperature 70 C are concentrated into phase
The medicinal extract for being 1.12~1.14 to density；
(8) 1-1.2g step (7) resulting medicinal extract is added into every liter of resulting seawater of step (2), and is uniformly mixed, i.e.,
Obtain object.
Enteromorpha special nutrient fluid obtained by a kind of above-mentioned preparation method.
The Enteromorpha of field acquisition is subjected to sterilization culture, experimental study table with nutrient solution of the invention in the lab
Bright, the Enteromorpha speed of growth is fast in sterilization incubation, Enteromorpha green in color, and the mature frond of sterilization culture is laid in LB
Solid culture primary surface is cultivated 10 days, does not there is varied bacteria growing.
The above is only a preferred embodiment of the present invention, it should be pointed out that: for the ordinary skill people of the art
For member, without departing from the principle of the present invention, several improvement can also be made, these improvement should be regarded as guarantor of the invention
Protect range.
Claims (9)
1. a kind of preparation method of Enteromorpha special nutrient fluid, it is characterised in that: the following steps are included:
(1) stand-by seawater is put into high-pressure steam sterilizing pan, is sterilized 20-30 minutes under the conditions of 121 DEG C；
(2) 0.04-0.05g NaNO is added in the seawater after every 1L sterilizing3、0.01g Na2HPO4·12H2O、0.02mg
MnCL2·4H2O、0.25-0.3mg FeSO4·7H2O、3.5-4mgNa20.2mgVB is added in EDTA after mixing1、
0.001mgVB12, 0.001mg biotin, be uniformly mixed；
(3) each component of following mass fraction: 32-37 parts of pennywort, 30-35 parts of cactus, 30-35 parts of onion, fish raw meat is weighed
27-33 parts careless, 21-28 parts of frutus cnidii, 21-26 parts of lophatherum gracile, 20-27 parts of dandelion, 18-25 parts of Prunella vulgaris, kiss-me 17-21
Part, 9-15 parts of peppermint oil, 14-20 parts of garlic, 12-18 parts of Cortex hibisus syriacus, 11-17 parts of the achene of Siberian cocklebur, 10-17 parts of kaempferia galanga, szechwan raspberry root or leaf
8-15 parts, 7-15 parts of Indian Epimeredi Herb, 6-10 parts of lotus nut, 6-10 parts of mulberry leaf, 5-9 parts of phellodendron bark, 5-9 parts of glutinous rice root；
(4) load weighted pennywort, cactus, onion are crushed and takes gained juice；
(5) load weighted cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic are crushed respectively, is added
Quality be cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 15 times of clear water, 20-30
After DEG C impregnating 5-6h, it is heated to the 1h that boils and be cooked by slow fire, fried liquid is taken out in separation；Continuing the addition quality into drug residue is
Cordate houttuynia, frutus cnidii, lophatherum gracile, dandelion, Prunella vulgaris, kiss-me, garlic gross mass 10 times of clear water, be heated to boiling simultaneously
It is cooked by slow fire 1h, merging fried liquid twice is extracting solution；
(6) load weighted Cortex hibisus syriacus, the achene of Siberian cocklebur, kaempferia galanga, szechwan raspberry root or leaf, Indian Epimeredi Herb, lotus nut, mulberry leaf, phellodendron bark, glutinous rice root are filled
Enter ball mill to be fully ground, crosses 80 meshes, obtain grounds travel；
(7) extracting solution and step (6) that load weighted peppermint oil, the resulting juice of step (4) and step (5) obtain are obtained
Grounds travel be placed in the extraction kettle in supercritical extract instrument, utilize carbon dioxide as medium and extracted, extraction kettle pressure
Power is 40Mpa, and extraction temperature is 45 DEG C, separator pressure 12Mpa, and separator temperature is 70 DEG C, extracts 2 hours, is extracted
Liquid；Extract liquor is concentrated in outer circulation inspissator, condition is concentrated are as follows: pressure 0.03Mpa, temperature 70 C are concentrated into relatively close
The medicinal extract that degree is 1.12~1.14；
(8) 1-1.2g step (7) resulting medicinal extract is added into every liter of resulting seawater of step (2), and is uniformly mixed to get mesh
Mark object.
2. the preparation method of Enteromorpha special nutrient fluid according to claim 1, it is characterised in that: in step (2), every 1L goes out
0.042g NaNO is added in culture seawater after bacterium3、0.01gNa2HPO4·12H2O、0.02mg MnCL2·4H2O、0.28mg
FeSO4·7H2O、3.7mgNa2EDTA is uniformly mixed, 0.2mgVB is then added1、0.001mgVB12, 0.001mg biotin, mix
It closes uniform.
3. the preparation method of Enteromorpha special nutrient fluid according to claim 1, it is characterised in that: in step (2), every 1L goes out
0.05g NaNO is added in culture seawater after bacterium3、0.01gNa2HPO4·12H2O、0.02mg MnCL2·4H2O、0.25mg
FeSO4·7H2O、3.8mgNa2EDTA is uniformly mixed, 0.2mgVB is then added1、0.001mgVB12, 0.001mg biotin, mix
It closes uniform.
4. the preparation method of Enteromorpha special nutrient fluid according to claim 1, it is characterised in that: in step (2), every 1L goes out
0.043g NaNO is added in culture seawater after bacterium3、0.01gNa2HPO4·12H2O、0.02mg MnCL2·4H2O、0.3mg
FeSO4·7H2O、4mgNa2EDTA is uniformly mixed, 0.2mgVB is then added1、0.001mgVB12, 0.001mg biotin, mixing
Uniformly.
5. the preparation method of Enteromorpha special nutrient fluid according to claim 1, it is characterised in that: in step (3), weigh with
The each component of lower mass fraction: 32 parts of pennywort, 33 parts of cactus, 30 parts of onion, 33 parts of cordate houttuynia, 28 parts of frutus cnidii, henon bamboo
25 parts of leaf, 27 parts of dandelion, 18 parts of Prunella vulgaris, 17 parts of kiss-me, 9 parts of peppermint oil, 15 parts of garlic, 12 parts of Cortex hibisus syriacus, the achene of Siberian cocklebur
17 parts, 16 parts of kaempferia galanga, 15 parts of szechwan raspberry root or leaf, 14 parts of Indian Epimeredi Herb, 10 parts of lotus nut, 10 parts of mulberry leaf, 9 parts of phellodendron bark, 8 parts of glutinous rice root.
6. the preparation method of Enteromorpha special nutrient fluid according to claim 1, it is characterised in that: in step (3), weigh with
The each component of lower mass fraction: 34 parts of pennywort, 30 parts of cactus, 33 parts of onion, 31 parts of cordate houttuynia, 25 parts of frutus cnidii, henon bamboo
26 parts of leaf, 25 parts of dandelion, 25 parts of Prunella vulgaris, 21 parts of kiss-me, 12 parts of peppermint oil, 14 parts of garlic, 15 parts of Cortex hibisus syriacus, the achene of Siberian cocklebur
11 parts, 14 parts of kaempferia galanga, 8 parts of szechwan raspberry root or leaf, 15 parts of Indian Epimeredi Herb, 9 parts of lotus nut, 9 parts of mulberry leaf, 8 parts of phellodendron bark, 9 parts of glutinous rice root.
7. the preparation method of Enteromorpha special nutrient fluid according to claim 1, it is characterised in that: in step (3), weigh with
The each component of lower mass fraction: 35 parts of pennywort, 35 parts of cactus, 32 parts of onion, 27 parts of cordate houttuynia, 23 parts of frutus cnidii, henon bamboo
22 parts of leaf, 20 parts of dandelion, 20 parts of Prunella vulgaris, 18 parts of kiss-me, 15 parts of peppermint oil, 17 parts of garlic, 18 parts of Cortex hibisus syriacus, the achene of Siberian cocklebur
12 parts, 17 parts of kaempferia galanga, 10 parts of szechwan raspberry root or leaf, 7 parts of Indian Epimeredi Herb, 6 parts of lotus nut, 6 parts of mulberry leaf, 5 parts of phellodendron bark, 7 parts of glutinous rice root.
8. the preparation method of Enteromorpha special nutrient fluid according to claim 1, it is characterised in that: in step (3), weigh with
The each component of lower mass fraction: 37 parts of pennywort, 32 parts of cactus, 35 parts of onion, 29 parts of cordate houttuynia, 21 parts of frutus cnidii, henon bamboo
21 parts of leaf, 22 parts of dandelion, 21 parts of Prunella vulgaris, 20 parts of kiss-me, 11 parts of peppermint oil, 20 parts of garlic, 16 parts of Cortex hibisus syriacus, the achene of Siberian cocklebur
15 parts, 10 parts of kaempferia galanga, 12 parts of szechwan raspberry root or leaf, 10 parts of Indian Epimeredi Herb, 8 parts of lotus nut, 8 parts of mulberry leaf, 7 parts of phellodendron bark, 5 parts of glutinous rice root.
9. the dedicated battalion of Enteromorpha obtained by a kind of preparation method of Enteromorpha special nutrient fluid of any of claims 1-8
Nutrient solution.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1416691A (en) * | 2001-11-03 | 2003-05-14 | 福州市海洋生物工程研究开发中心 | Micro algae culturing liquid and its prepn process |
| CN203015575U (en) * | 2012-12-30 | 2013-06-26 | 福建海兴保健食品有限公司 | Novel enteromorpha prolifera cultivating tool |
| CN103609425A (en) * | 2013-12-10 | 2014-03-05 | 上海海洋大学 | Sterile cultivation method for enteromorpha |
| CN104230512A (en) * | 2013-06-23 | 2014-12-24 | 青岛道合生物科技有限公司 | Enteromorpha prolifera amino acid plant nutrient agent |
| JP2015107061A (en) * | 2013-12-03 | 2015-06-11 | 新日鐵住金株式会社 | Method for growing sea weed |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1416691A (en) * | 2001-11-03 | 2003-05-14 | 福州市海洋生物工程研究开发中心 | Micro algae culturing liquid and its prepn process |
| CN203015575U (en) * | 2012-12-30 | 2013-06-26 | 福建海兴保健食品有限公司 | Novel enteromorpha prolifera cultivating tool |
| CN104230512A (en) * | 2013-06-23 | 2014-12-24 | 青岛道合生物科技有限公司 | Enteromorpha prolifera amino acid plant nutrient agent |
| JP2015107061A (en) * | 2013-12-03 | 2015-06-11 | 新日鐵住金株式会社 | Method for growing sea weed |
| CN103609425A (en) * | 2013-12-10 | 2014-03-05 | 上海海洋大学 | Sterile cultivation method for enteromorpha |
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