CN1243792C - Method for extracting algin from fake valoniopsis pachynema - Google Patents
Method for extracting algin from fake valoniopsis pachynema Download PDFInfo
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- CN1243792C CN1243792C CN 200410012925 CN200410012925A CN1243792C CN 1243792 C CN1243792 C CN 1243792C CN 200410012925 CN200410012925 CN 200410012925 CN 200410012925 A CN200410012925 A CN 200410012925A CN 1243792 C CN1243792 C CN 1243792C
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- valoniopsis pachynema
- fake valoniopsis
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- pachynema
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Abstract
The present invention discloses a method for extracting scytonema arcangelii elements from scytonema arcangelii, which has the steps that a scytonema arcangelii culture solution is firstly prepared from dipotassium hydrogen phosphate, magnesium sulfate, calcium chloride, citric acid, ferric ammonium citrate, disodium edetate dihydrate (EDTA), sodium carbonate, and a trace element solution A5 according to proportions; secondly, the scytonema arcangelii is cultivated, the conditions of temperature, light intensity, light quality and ventilation of the culture solution are controlled; thirdly, scytonema arcangelii powder is prepared; fourthly, the scytonema arcangelii elements are extracted; finally, the scytonema arcangelii elements are obtained and are dried. The present invention has the advantages of easy implementation, convenient operation, simple equipment, high yield, high product quality, and high safety and reliability. The present invention is suitable for scale production of industrialization.
Description
Technical field
The present invention relates to the biotechnology and the utilization of resources and Sustainable development field, more specifically relate to a kind of method of utilizing fake valoniopsis pachynema to extract fake valoniopsis pachynema.
Background technology
At present, the utilization of fake valoniopsis pachynema generally only rested on carry out on the artificial algae skinning,, say nothing of and from fake valoniopsis pachynema, extract fake valoniopsis pachynema and be used other application and developments of fake valoniopsis pachynema still blank out almost.
Fake valoniopsis pachynema (scytonemin) is a class tawny, and fat-soluble dimer pigment, molecular weight are 544Da, and structure is indoles or phenol subunit, and its maximum absorption band is at 384nm.A lot of desert blue-green algaes all contain fake valoniopsis pachynema, and this fake valoniopsis pachynema has the characteristic of anti-ultraviolet radiation, thereby the protection fake valoniopsis pachynema is avoided the injury of UV-light, for the normal growth of fake valoniopsis pachynema creates conditions.This ability with extremely strong anti-ultraviolet radiation of fake valoniopsis pachynema can be used for cosmetic industry.And at present, the pigment that China uses mainly is the chemosynthesis pigment, and the life-time service synthetic colour causes damage to human body skin, thereby is unfavorable for human health.
At present, domestic extraction to fake valoniopsis pachynema sets foot in very few, and its extracting method is loaded down with trivial details, the cost height.
Summary of the invention
The object of the present invention is to provide a kind of method of extracting fake valoniopsis pachynema from fake valoniopsis pachynema, this method is easy, and is easy to operate, and equipment is simple, safe and reliable, and cost is low, and good product quality is suitable for suitability for industrialized production.
The concrete steps of extracting fake valoniopsis pachynema from fake valoniopsis pachynema are as follows:
1, the preparation of nutrient solution: every premium on currency adds dipotassium hydrogen phosphate 0.02-0.05g, sal epsom 0.06-0.09g, calcium chloride 0.02-0.05g, citric acid 0.004-0.008g, ferric ammonium citrate 0.004-0.008g, disodium EDTA (EDTA) 0.001-0.003g, yellow soda ash 0.01-0.04g, trace element solution A50.5-2ml;
2, the cultivation of fake valoniopsis pachynema:
(1) one-level is cultivated: insert triangular flask (150-200ml) by the test tube original seed, static cultivation or aerated culture during aerated culture, are regulated the air-flow size at 1.5-3.5L.min
-1, air-flow carries out aseptically process through cotton ball, and light intensity is controlled at 65-85 μ E.m
-2.s
-1, temperature is controlled at 22-35 ℃.After cultivating 5-10 days, be transferred to secondary and cultivate.
(2) secondary is cultivated: the culture of one-level being cultivated gained is linked into culturing bottle (500-1000ml), and aerated culture is regulated the air-flow size at 2.5-5L.min
-1, aseptically process, light intensity are controlled at 75-95 μ E.m
-2.s
-1, temperature is controlled at 22-35 ℃.After cultivating 5-10 days, be transferred to three grades of cultivations.
(3) three grades of cultivations: the culture of secondary being cultivated gained is linked into culturing bottle (5000-20000ml), and aerated culture is regulated the air-flow size at 3-6L.min
-1, aseptically process, light intensity are controlled at 70-100 μ E.m
-2.s
-1, temperature is controlled at 22-35 ℃.After cultivating 5-10 days, be the inoculation provenance that continues to cultivate.
(4) continue to cultivate: adopt the algae kind after three grades of cultivations, in open runway circulating culturing pool, cultivate in a large number in the ratio that inserts 0.2-0.5g (weight in wet base) fake valoniopsis pachynema in every liter of nutrient solution.Adopt glass temperature canopy controlled temperature at 28-30 ℃, utilize natural light to carry out illumination, light intensity is controlled at 80-120 μ E.m
-2.s
-1, adopt impeller or submersible pump to stir substratum.In 3-6 days before results, twice employing every day UV-B irradiation, light intensity is controlled at 120-160 μ E.m
-2.s
-1, each time was at 5-10 minute.
3, the preparation of fake valoniopsis pachynema algae powder:
The fake valoniopsis pachynema of cultivation after 10-20 days is extracted into filtration unit with pump---vibrate on the inclined-plane sieve, after sieving through the vibration inclined-plane, receive to such an extent that the fake valoniopsis pachynema algae is starched, then that the algae slurry is centrifugal through whizzer, obtain fake valoniopsis pachynema algae mud.After concentrated, dehydration, drying, pulverizing, obtain fake valoniopsis pachynema algae powder.
4, the extraction of fake valoniopsis pachynema (scytonemin):
With the algae powder under 28-32 ℃, add anhydrous methanol (the algae powder: methyl alcohol=1: 8-12, W/V), utilize agitator ceaselessly to stir extracting 30min, extracting 2-3 time is up to the extract very slight color.In super-magnum centrifuge, the centrifugal 10-15min of 5000-8000g collects supernatant liquor, add ethyl acetate (its volume be the used methyl alcohol cumulative volume in front 1/10th) then in throw out, 28-32 ℃ is continued extracting down, centrifugal, collect supernatant liquor, merge with the methyl alcohol extract.Add the long-pending distilled water of one halfbody then in the extract after merging, placed 10-15 minute down in-5--10 ℃, promptly visible fake valoniopsis pachynema (scytonemin) precipitation is separated out.
5, the results of fake valoniopsis pachynema are with dry: with the above-mentioned centrifugal 10-15min of solution 5000-8000g in super-magnum centrifuge that contains fake valoniopsis pachynema, collecting precipitation thing.-60--110 ℃ of following lyophilize.
The present invention compared with prior art has the following advantages and significant effect: easy to implement the method, easy to operate, equipment simple, economic quick, high, the good product quality, safe and reliable of output, is applicable to commercial scale production.
Embodiment
The concrete implementation step of extracting fake valoniopsis pachynema is as follows:
1, the preparation of nutrient solution: every premium on currency adds nitre dipotassium hydrogen phosphate 0.04g, sal epsom 0.075g, calcium chloride 0.036g, citric acid 0.006g, ferric ammonium citrate 0.006g, disodium EDTA (EDTA) 0.001g, yellow soda ash 0.02g, trace element solution A
51ml;
2, the cultivation of fake valoniopsis pachynema:
(1) one-level is cultivated: insert triangular flask (150-200ml) by the test tube original seed, static cultivation or aerated culture during aerated culture, are regulated the air-flow size at 2.5L.min
-1, air-flow carries out aseptically process through cotton ball, and light intensity is controlled at 70 μ E.m
-2.s
-1, temperature is controlled at 30 ℃.After cultivating 7-10 days, be transferred to secondary and cultivate.
(2) secondary is cultivated: the culture of one-level being cultivated gained is linked into culturing bottle (500-1000ml), and aerated culture is regulated the air-flow size at 3L.min
-1, the aseptically process method is the same, and light intensity is controlled at 80 μ E.m
-2.s
-1, temperature is controlled at 30 ℃.After cultivating 7-10 days, be transferred to three grades of cultivations.
(3) three grades of cultivations: the culture of secondary being cultivated gained is linked into culturing bottle (5000-20000ml), and aerated culture is regulated the air-flow size at 5L.min
-1, the aseptically process method is the same, and light intensity is controlled at 90 μ E.m
-2.s
-1, temperature is controlled at 30 ℃.After cultivating 7-10 days, be the inoculation provenance that continues to cultivate.
(4) continue to cultivate: adopt the algae kind after three grades of cultivations, be linked in the open runway circulating culturing pool, cultivate in a large number in the ratio of every liter of nutrient solution 0.3-0.4g weight in wet base.Adopt glass temperature canopy controlled temperature at 30 ℃, utilize natural light to carry out illumination, light intensity is controlled at 100-120 μ E.m
-2.s
-1, adopt impeller or submersible pump to stir substratum.In 3-6 days before results, twice employing every day UV-B irradiation, light intensity is controlled at 140-160 μ E.m
-2.s
-1, each time was at 5-10 minute.
3, the preparation of fake valoniopsis pachynema algae powder:
The fake valoniopsis pachynema of cultivation after 10-20 days is extracted into filtration unit with pump---vibrate on the inclined-plane sieve, after sieving through the vibration inclined-plane, receive to such an extent that the fake valoniopsis pachynema algae is starched, then that the algae slurry is centrifugal through whizzer, obtain fake valoniopsis pachynema algae mud.After concentrated, dehydration, drying, pulverizing (common process), obtain fake valoniopsis pachynema algae powder.
4, the extraction of fake valoniopsis pachynema (scytonemin):
With the algae powder under 30 ℃, add anhydrous methanol (the algae powder: methyl alcohol=1: 10, W/V), utilize agitator ceaselessly to stir extracting 30min, extracting 2-3 time is until the extract very slight color.In super-magnum centrifuge, the centrifugal 10-15min of 5000-8000g collects supernatant liquor, add ethyl acetate (its volume be the used methyl alcohol cumulative volume in front 1/10th) then in throw out, 30 ℃ are continued extractings down, centrifugal, collect supernatant liquor, merge with the methyl alcohol extract.Add the long-pending distilled water of one halfbody then in the extract after merging, placed 10-15 minute down in-5 ℃, promptly visible fake valoniopsis pachynema precipitation is separated out.
5, the results of fake valoniopsis pachynema are with dry: with the above-mentioned centrifugal 10-15min of solution 5000-8000g in super-magnum centrifuge that contains fake valoniopsis pachynema, collecting precipitation thing.-100 ℃ of following lyophilizes.
Claims (1)
1, a kind of method of extracting fake valoniopsis pachynema from fake valoniopsis pachynema comprises the steps:
The preparation of A, nutrient solution: every premium on currency adds dipotassium hydrogen phosphate 0.02-0.05g, sal epsom 0.06-0.09g, calcium chloride 0.02-0.05g, citric acid 0.004-0.008g, ferric ammonium citrate 0.004-0.008g, disodium EDTA 0.001-0.003g, yellow soda ash 0.01-0.04g, trace element solution A
50.5-2ml;
The cultivation of B, fake valoniopsis pachynema: at first be that one-level is cultivated, insert in the triangular flask that aerated culture is regulated air-flow at 1.5-3.5L.min by the test tube original seed
-1, air-flow carries out aseptically process through cotton ball, and light intensity is controlled at 65-85 μ E.m
-2.s
-1, temperature is controlled at 22-35 ℃, after cultivating 5-10 days, is transferred to secondary and cultivates; Next is that secondary is cultivated, and the culture of one-level being cultivated gained is linked in the culturing bottle, and aerated culture is regulated air-flow at 2.5-5L.min
-1, aseptically process, light intensity are controlled at 75-95 μ E.m
-2.s
-1, temperature is controlled at 22-35 ℃, after cultivating 5-10 days, is transferred to three grades of cultivations; The 3rd is three grades of cultivations, and the culture of secondary being cultivated gained is linked in the culturing bottle, and aerated culture is regulated air-flow at 3-6L.min
-1, aseptically process, light intensity are controlled at 70-100 μ E.m
-2.s
-1, temperature is controlled at 22-35 ℃, after cultivating 5-10 days, is the inoculation provenance that continues to cultivate; The 4th is continue to cultivate, and adopts the algae kind after three grades of cultivations, is linked in the open runway circulating culturing pool in the ratio of every liter of nutrient solution 0.2-0.5g weight in wet base, cultivate, adopt glass temperature canopy controlled temperature at 28-30 ℃, utilize natural light to carry out illumination, light intensity is controlled at 80-120 μ E.m
-2.s
-1, adopt impeller or submersible pump to stir substratum, in 3-6 days before results, twice employing every day UV-B irradiation, light intensity is controlled at 120-160 μ E.m
-2.s
-1, each time was at 5-10 minute;
The preparation of C, fake valoniopsis pachynema algae powder: the fake valoniopsis pachynema that will cultivate after 10-20 days is extracted into filtration unit with pump---on the vibration inclined-plane sieve, behind vibration inclined-plane sieve, receive to such an extent that the fake valoniopsis pachynema algae is starched, then that the algae slurry is centrifugal through whizzer, obtain fake valoniopsis pachynema algae mud, after concentrated, dehydration, drying, pulverizing, obtain fake valoniopsis pachynema algae powder;
The extraction of D, fake valoniopsis pachynema: with the algae powder under 28-32 ℃, the anhydrous methanol that adds 8-12 times of volume, utilize agitator ceaselessly to stir extracting 30min, extracting 2-3 time, up to the extract very slight color, in whizzer, the centrifugal 10-15min of 5000-8000g collects supernatant liquor, in throw out, add ethyl acetate then, 28-32 ℃ is continued extracting down, centrifugal, collects supernatant liquor, merge with the methyl alcohol extract, add the long-pending distilled water of one halfbody then in the extract after merging, in-5--10 ℃ place 10-15 minute down after, the fake valoniopsis pachynema precipitation is just separated out;
The results of E, fake valoniopsis pachynema are with dry: with the above-mentioned centrifugal 10-15min of solution 5000-8000g in whizzer that contains fake valoniopsis pachynema, collecting precipitation thing ,-60--110 ℃ of following lyophilize.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410012925 CN1243792C (en) | 2004-03-27 | 2004-03-27 | Method for extracting algin from fake valoniopsis pachynema |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410012925 CN1243792C (en) | 2004-03-27 | 2004-03-27 | Method for extracting algin from fake valoniopsis pachynema |
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| CN1563205A CN1563205A (en) | 2005-01-12 |
| CN1243792C true CN1243792C (en) | 2006-03-01 |
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| CN 200410012925 Expired - Fee Related CN1243792C (en) | 2004-03-27 | 2004-03-27 | Method for extracting algin from fake valoniopsis pachynema |
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| CN101816685B (en) * | 2010-05-05 | 2012-01-11 | 暨南大学 | Hydroxyl-containing acid extract for preventing and treating prostate cancer, and extraction method and application thereof |
| WO2020128112A1 (en) * | 2018-12-21 | 2020-06-25 | Ciimar - Centro De Investigação Marinha E Ambiental | Ultra-violet absorbing compounds |
| CN117682643A (en) * | 2024-02-02 | 2024-03-12 | 杭州楠大环保科技有限公司 | Method for preparing microalgae flocculant by using microalgae |
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Legal Events
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|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Da-chuan Panyu in Guangzhou City Panyu Da-chuan Feed Co., Ltd. Assignor: Institute of Hydrobiology, Chinese Academy of Sciences Contract fulfillment period: 2007.2.1 to 2012.1.31 contract change Contract record no.: 2008420000013 Denomination of invention: Method for extracting algin from fake valoniopsis pachynema Granted publication date: 20060301 License type: Exclusive license Record date: 2008.7.7 |
|
| LIC | Patent licence contract for exploitation submitted for record |
Free format text: EXCLUSIVE LICENCE; TIME LIMIT OF IMPLEMENTING CONTACT: 2007.2.1 TO 2012.1.31 Name of requester: GUANGZHOU CITY FANYU DISTRICT DACHUAN FEEDSTUFF C Effective date: 20080707 |
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| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20060301 Termination date: 20130327 |