CN102827303A - Method for producing spirulina polysaccharide by fresh spirulina - Google Patents
Method for producing spirulina polysaccharide by fresh spirulina Download PDFInfo
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Abstract
The invention relates to a method for extracting spirulina polysaccharide, which comprises the steps of: filtering out nutrient solution, cleaning, extracting, separating, concentrating, alcohol precipitating, washing, drying, crushing, screening and the like. The method takes spirulina mud as raw material, so that a device used during spirulina drying process can be omitted, and the energy consumption is reduced. Compared with a method for extracting the spirulina polysaccharide with spirulina powder, the method avoids the damage to the polysaccharide structure in the high-temperature drying process of the spirulina powder on one hand, and avoids the phenomenon of getting partially burned because of over-heating near fire on the other hand, thus improving the yield and the quality of the spirulina polysaccharide, reducing the dosage of alcohol needed by decoloration, greatly reducing the production cost and improving the product quality. The fresh spirulina is taken as raw material, and chemical reagent is not needed to be added into the raw material for pretreatment, so that chemical residue can be avoided.
Description
Technical field
The invention belongs to algae separation and purification biological extraction technical field; Relate to a kind of being applicable to tenaculat Habenaria through poach-extraction separation-concentrated method of processing the spirulina polysaccharide product, particularly a kind of new method for extracting that is suitable for becoming the higher-grade spirulina polysaccharide with the fresh spirulina separation and purification.
Background technology
Tenaculat Habenaria (Spirulina) is a kind of photosynthetic oxygen evolution, spiral filamentous cyanobacteria; Belong to the algae guiding principle Spirullina of quivering, be described as " optimal food of 21 century " and " optimum medicines of human 21 century " by the Food and Argriculture OrganizationFAO and the World Health Organization respectively.In China, classified as protective foods.Investigation shows that the year consumption of 10 years from now on internal screw algaes will reach ten thousand tons of l, and product relates to medicine, protective foods, fields such as makeup.
Spirulina polysaccharide is a kind of acid heteroglycan; It is important physical activeconstituents very in the tenaculat Habenaria; Have unique effect at aspects such as antitumor, antiviral, hypoglycemic, anti-ageing, radioprotective, anticoagulation and raising immunizing power, become the focus in current tenaculat Habenaria development research field.(Deng Zhongri, You Peiqing, Wang Xiaobing. spirulina polysaccharide progress Hunan City University journal the 14th the 3rd phase 63-65. of volume of September in 2005) more about the functional study of spirulina polysaccharide both at home and abroad.There is report research to point out; Spirulina polysaccharide can significantly suppress propagation and ascites hepatoma cells DNA, RNA and proteinic the synthesizing of murine osteosarcoma knurl 180 cells; Restraining effect in 3～24 hours in time prolongation and improve, different cancer cells is different to the susceptibility of spirulina polysaccharide.Guangzhou Medical College Ceng Bohang etc. find during to the result of treatment of leukemia patient at the research spirulina polysaccharide; Spirulina polysaccharide influences the active nothing of normal people's nk cell (NK cell) external; Send out, recur phase patient peripheral blood NK cell activity but can improve in various degree just, and under the activation of interleukin-2, can improve leukemia patient peripheral blood LAK (LAK cell) activity.To being in paracmastic Patients with Acute Leukemia, spirulina polysaccharide can make its NK cytoactive recover normal.It is relevant with body band warty attitude that spirulina polysaccharide improves the effect of tumour patient NK cytoactive, when tumor load acts on obviously (2000 the 6th phases of " Chinese marine drug " magazine (total the 78th phase)) when little (catabasis).Zhang Chengwu, Pang Qishen, Xuchang Shao etc. discover, spirulina polysaccharide and gp all have radioprotective, anti-oxidant, enhance immunity power, promote lymphocyte transformation and anticancer proliferation function.In to the research of spirulina polysaccharide radioprotective mechanism, find its radiation resistance and the activity of endonuclease and reinforcing effect relevant (Journal of Nutrition, 1996, the l8 (3): 327. of dna damage repair enzyme; Acta Genetica Sinica, 1988,15 (5): 374.; China's radiological medicine and protection magazine, 1996, (3): 136 1 138.).Spirulina polysaccharide is the pair cell nontoxicity not only; And having the myelocytic multiplication capacity of enhance bone, hemopoietic improves leucocyte level; Improve the phagocytic rate and the phagocytic index of scavenger cell; The restraining effect of lymphocytic quantity of increase T and elimination immunosuppressor etc. (Xue Zhiyong, the research of spirulina polysaccharide and application Shandong food science and technology, 2004.No.8.5-6).Simultaneously, spirulina polysaccharide and gp can also improve lipid metabolism, reduce blood plasma cholesterol level, and gastric and duodenal ulcer is had certain auxiliary curative effect.
At present; China's tenaculat Habenaria industrialization is produced and is succeedd; Its produce and evolution in outstanding problem be: the tenaculat Habenaria deep processed product is in the junior stage, and domesticly rests on the laboratory study stage about spirulina polysaccharide extraction and purification technology more, and it is less to be applied to production practice.Introducing in " the extraction purifying of natural spiral seaweed polysaccharide and hypoglycemic activity experimental study " (processing of farm products, in January, 2007) like Jia Shaoting, Yin Wenzheng, is that raw material extracts spirulina polysaccharide and (gets spirulina powder with the spirulina powder; Adding massfraction by solid-to-liquid ratio 1:10 is 0.1% sodium pyrophosphate solution broken wall, adopts hot-water extraction, is 1:9 by solid-to-liquid ratio; Temperature 80 ~ C; Extracting 6h, extracting 2 times is through rotating speed 4000r/min centrifuging and taking supernatant; Constant volume is to 500mL, and it is subsequent use to put into refrigerator.Get above-mentioned supernatant 50mL, be concentrated into 20%, add ethanol doubly the adding liquid concentrator by 4, place 12h at 4 ℃ of refrigerators and take out, with the centrifugal 20min of 4000r/min rotating speed.With zero(ppm) water Crude polysaccharides fully being dissolved, is that 1:8 processes solution by solid-to-liquid ratio, and adjust pH is 7.0; The neutral protease that adds people's Crude polysaccharides quality 3% respectively; After temperature is incubated 2.5h for 50 ℃, treats complete enzymolysis, adopt the Sevage method to carry out deproteinated and handle; Extract Crude polysaccharides, calculate polysaccharide content).And for example Pan Qiuwen, height are eastwards, to contain the spirulina polysaccharide process for extracting of Hailin etc. in " Study on extraction of spirulina polysaccharide " (medical Leader the 23rd the 9th phase of volume of September in 2004), mentioning also be to be that raw material extracts polysaccharide with the spirulina powder.(tenaculat Habenaria pulverized makes spirulina powder, behind the potass extraction, extracting solution through the centrifugal 20min of 3000rmin, is stayed supernatant, residue is carried out 2 times extract, with extracting solution through the centrifugal 20min of 3000rmin-.Two times centrifugal gained supernatant is merged, adopt behind the Sevage method Deproteinization the Deproteinization polysaccharide, after dialysing with dialyzate with 3 times of ethanol sedimentations, throw out is used Vanadium Pentoxide in FLAKES vacuum-drying, promptly get dry product SP).Also having the process for extracting in " the orthogonal experiment optimization of the research of spirulina plalensis polysaccharide extracting process---trichoroacetic acid(TCA) (TCA) method " such as Sun Yuanzheng, Zhang Xuecheng, Ji Lei also is to be that raw material (ocean science/2oo7/the 31st/the 4 phase of volume) takes by weighing the 100g spirulina powder with the spirulina powder; Add 800mL95 ethanol, soaked overnight; The centrifugal 20rain of 3600r/rain, 37 ℃ of baking dry algae powders add distilled water by the 1:12 mass ratio, with NaOH the pH value of solution value are transferred to 10; 80 ℃ of water-bath 6h; The centrifugal 3Omin of 3600r/rain gets supernatant, add 10% hydrochloric acid supernatant pH value transferred to 7.0, after dropwise add 5% trichloroacetic acid precipitation albumen, 4 ℃ are spent the night; The centrifugal 20min of 3 600r/rain, supernatant add 5% trichloroacetic acid precipitation albumen again, leave standstill 3h; The centrifugal 20min of 3600r/rain, on reset and add 5 times of volumes, 95% ethanol sedimentation polysaccharide, 4 ℃ are spent the night; The centrifugal 2Omin of 3600r/min gets deposition, with washing with acetone deposition twice, lyophilize.In a word; Because the polysaccharide of different types of tenaculat Habenaria preparation is different, the difference of extraction separation purification process in addition, the polysaccharide of same tenaculat Habenaria preparation is also different; Therefore; The kind of spirulina polysaccharide is numerous and diverse theoretically, does not have the polysaccharide kind title of standard basically, hinders polysaccharide further research and application.(the 14th the 3rd phase of volume of Hunan City University's journal (natural science edition), in September, 2005)
Since now prepared in laboratory spirulina polysaccharide multiselect with spirulina powder as raw material, and tenaculat Habenaria in drying course because the high temperature one side can be destroyed polysaccharide structures, can produce the phenomenon of adustion on the other hand, influence the output capacity and the quality of polysaccharide.Moreover the structure of spirulina polysaccharide itself belongs to gp, and component is more; If refining and improper its active function that reduces possibly of purifying (referring to Deproteinization) method; The pharmaceutical chemicals that laboratory extractive technique complicated steps relates to is many, lacks unified standard, and the incompatibility suitability for industrialized production needs; Remove chemical residual, improve active polysaccharide, to enlarge polysaccharide industrial scale difficulty very big, thereby limited the research and development of refining spirulina polysaccharide.
At present, the tenaculat Habenaria deep processed product is in the junior stage, and domestic relevant spirulina polysaccharide extraction and purification technology rests on the laboratory study stage more, and it is less to be applied to production practice.Especially be that the direct large-scale production spirulina polysaccharide of raw material also exists following problems to have to be solved with the fresh spirulina:
1, the place and the equipment of the technology of the Biochemical Lab of existing most research and teaching units shortage cultivation tenaculat Habenaria and big area, industrialized culture.Therefore, fresh spirulina can not be self-sufficient, limited the further test and the research of extracting polysaccharide with fresh spirulina.Be confined to extract polysaccharide as raw material with the spirulina powder that can buy on the market.
2, vast tenaculat Habenaria manufacturing enterprise carries on the work with the form of spirulina powder, and product is in the junior stage, and lacks the deep process technology and the talent that extract polysaccharide with tenaculat Habenaria.
3, through the contrast to using the spiral algae powder and using fresh spirulina to extract polysaccharide, the clear and definite difference that has some extracting parameters technically as raw material.The broken wall, solid-liquid ratio, extraction temperature, extraction time, the alcohol that mainly are reflected in the spirulina cells wall are analysed aspects such as decolouring.The application improves through following (C-I) and uses fresh spirulina to become possibility as raw material extraction polysaccharide.
Summary of the invention
The object of the present invention is to provide a kind of method of using fresh spirulina to extract spirulina polysaccharide, to overcome above-mentioned existing in prior technology weak point.
The objective of the invention is to accomplish through following technical proposals
Algae mud directly adds hydro-thermal and boils the method for extracting spirulina polysaccharide, contains following technological process:
1, a kind of method of producing spirulina polysaccharide with fresh spirulina,
A, elimination nutrient solution: the tenaculat Habenaria of normal growth in the cultivation pool is picked up, be poured on elimination nutrient solution on the sieve,
B, cleaning: the tenaculat Habenaria algae mud of elimination moisture and nutrient solution is cleaned with tap water, and filter is done, and is 3 times repeatedly, subsequent use;
C, extraction: the tenaculat Habenaria algae mud for preparing is put into the stainless steel heating kettle, add water and heating, extract for some time;
D, separation: the mixture after will extracting injects whizzer;
E, concentrated: the liquid portion heating is concentrated, cool off subsequent use;
F, alcohol are analysed: cooled liquid concentrator is put into stainless steel cask add ethanol, abandoning supernatant is taken out throw out;
G, washing: throw out is continued to use washing with alcohol, the filter of the throw out after the washing is done;
H, drying: the throw out that will filter after doing is put into the vacuum drier inner drying;
I, pulverize, sieve: dried polysaccharide is carried out crushing screening (60-80 order).
Wherein, among the step C, the consumption of extracting solution (water) is tenaculat Habenaria 2-3 times, is heated to and boils; Extracted 3-4 hour, among the step D, it is centrifugal that separating machine is that disc centrifuge or three whizzers cooperate tubular-bowl centrifuge to carry out, among the step E; To original volume 1/3-1/5, in the step F, ethanol concn is 95% with liquid concentration, and filling rate is 3-4 times of solution after concentrating; Among the step G, scavenging solution is 95%-absolute ethyl alcohol, and number of times is 2-3 time.
The invention has the beneficial effects as follows, adopt fresh spirulina, compared following some benefit with the described process for extracting of technical background as the raw material that extracts spirulina polysaccharide:
(1) reduced the tenaculat Habenaria drying course equipment addition, reduced energy consumption.
(2) compare with the method for extracting polysaccharide with spirulina powder; One aspect of the present invention has been avoided spiral algae powder destruction to polysaccharide structures in the hyperthermia drying process; Avoided the phenomenon of adustion on the other hand; Improve the output capacity and the quality of polysaccharide, reduce the required alcohol consumption of decolouring, raising quality significantly can reduce production costs.(3) the present invention adopts fresh spirulina as raw material, need not to add any chemical reagent and carries out pre-treatment, has avoided chemical residual.
Description of drawings
Fig. 1 is technological process of production figure.
Embodiment
Embodiment one
The tenaculat Habenaria of normal growth in the cultivation pool is picked up, be poured on elimination nutrient solution on the wide 1 meter long 5 meters aperture 200 purpose sieves.When building up to 25 kilograms of left and right sides, the tenaculat Habenaria algae mud of elimination moisture and nutrient solution was cleaned 5 minutes with tap water, filter is done, 3 times repeatedly.
The tenaculat Habenaria algae mud for preparing is put into 65 centimetres high 60 centimetres of stainless steel heating kettles of diameter, add 2 times in water (50 kilograms) and boil, stir and extracted 3 hours.(the stainless steel heating kettle can be according to industrial scale sized and quantity, and should possess agitating function, compares with the technology of extracting polysaccharide with spirulina powder, has saved the middle-chain of algae powder oven dry, has practiced thrift cost and reduced the algae burgyization that drying course causes).
To extract mixture after the cooling and inject disc centrifuge (we select for use be the DBP-50 type that oasis, Nanjing machine plant produces) and carry out solid-liquid separation, and remove residue, (concrete operations are carried out by the regulation in producer's working instructions) liquid portion keeps subsequent use.
Liquid portion is put into above-mentioned Stainless Steel Kettle internal heating or vacuum distilling apparatus is removed excessive moisture, be concentrated into 1/3 volume after (cooling) subsequent use.
Cooled liquid concentrator is put into 50 centimetres high stainless steel casks of 50 centimetres of diameter, add 4 times of 95% ethanol, treat condensation prod post precipitation (needing 24 hours usually) removal supernatant, throw out (spirulina polysaccharide) is taken out the elimination unnecessary alcohol.Continue with 95% washing with alcohol 2 times, to supernatant limpid colourless till, the filter of the throw out after the washing is dried.Put into the vacuum drier inner drying, dried polysaccharide is carried out crushing screening (60-80 order).
Embodiment two
Production process is as shown in Figure 1.
1, be raw material with algae mud, step such as embodiment one
C---25 kilograms in the tenaculat Habenaria algae mud for preparing is put into 65 centimetres high 60 centimetres of stainless steel heating kettles of diameter, add water and boil for 75 kilograms, stir and extracted 4 hours.
D---will extract mixture injection disc centrifuge after the cooling and carry out solid-liquid separation, and remove residue, liquid portion keeps subsequent use.Altogether 35.6 kilograms of polysaccharide extraction liquids.
E---liquid portion is put into above-mentioned Stainless Steel Kettle internal heating (or vacuum distilling apparatus) removes excessive moisture, be concentrated into 1/5 volume after (cooling) subsequent use.Altogether 7.1 kilograms of polysaccharide liquid concentrators.
F---cooled liquid concentrator is put into 50 centimetres high stainless steel casks of 50 centimetres of diameter, add 28 liters of absolute ethyl alcohols, precipitate and remove supernatant after 24 hours, throw out (spirulina polysaccharide) is taken out the elimination unnecessary alcohol.
G---continue use absolute ethanol washing, to supernatant limpid colourless till, the throw out after (each 28 liters, totally 3 times) will washs is filtered dried.
H---the polysaccharide precipitation thing was put into the vacuum drier inner drying 24 hours.
I---dried polysaccharide is carried out crushing screening 80 orders.(being total to such an extent that polysaccharide powder 236 restrains)
1, be raw material with the algae powder, step such as embodiment one
C---2.5 kilograms of spiral algae powders are put into 65 centimetres high 60 centimetres of stainless steel heating kettles of diameter, add water and boil for 100 kilograms, stir and extracted 4 hours.
D---will extract mixture injection disc centrifuge after the cooling and carry out solid-liquid separation, and remove residue, liquid portion keeps subsequent use.Altogether 32.3 kilograms of polysaccharide extraction liquids.
E---liquid portion is put into above-mentioned Stainless Steel Kettle internal heating or vacuum distilling apparatus is removed excessive moisture, be concentrated into 1/5 volume after (cooling) subsequent use.Altogether 6.5 kilograms of polysaccharide liquid concentrators.
F---cooled liquid concentrator is put into 50 centimetres high stainless steel casks of 50 centimetres of diameter, add 26 liters of absolute ethyl alcohols, precipitate and remove supernatant after 24 hours, throw out (spirulina polysaccharide) is taken out the elimination unnecessary alcohol.
G---continue use absolute ethanol washing, to supernatant limpid colourless till, the throw out after (each 26, totally 5 times) will wash is filtered dried.
H-polysaccharide precipitation the thing was put into the vacuum drier inner drying 24 hours.
I---dried polysaccharide is carried out crushing screening 80 orders.(being total to such an extent that polysaccharide powder 192 restrains)
Two kinds of raw material contrast experiment tables:
10kg algae mud gets 1kg algae powder, can obviously find out from last table, extracts polysaccharide with algae mud as raw material, and than using the algae powder to exceed 23% as raw material, used amount of alcohol is that raw material lacks 40% than the algae powder on the output.Difference is very remarkable between the two.
Two kinds of raw materials, 4 groups of spirulina polysaccharide output such as following tables that repeat gained:
Variance analysis
| Source of variation | df | SS | MS | F | F 0.01 |
| Between processing | 1 | 3890.9431 | 3890.9431 | 6012.16 | 13.74 |
| Error | 6 | 3.8831 | 0.6472 | ||
| Total variation | 7 | 3894.8262 |
Remarkable through the variance analysis result for extremely.
Claims (1)
1. method of producing spirulina polysaccharide with fresh spirulina may further comprise the steps:
A, elimination nutrient solution: the tenaculat Habenaria of normal growth in the cultivation pool is picked up, be poured on elimination nutrient solution on the sieve,
B, cleaning: the tenaculat Habenaria algae mud of elimination moisture and nutrient solution is cleaned with tap water, and filter is done, and is 3 times repeatedly, subsequent use;
C, extraction: the tenaculat Habenaria algae mud for preparing is put into the stainless steel heating kettle, add water and heating, extract for some time;
D, separation: the mixture after will extracting injects whizzer;
E, concentrated: the liquid portion heating is concentrated, cool off subsequent use;
F, alcohol are analysed: cooled liquid concentrator is put into stainless steel cask add ethanol, abandoning supernatant is taken out throw out;
G, washing: throw out is continued to use washing with alcohol, the filter of the throw out after the washing is done;
H, drying: the throw out that will filter after doing is put into the vacuum drier inner drying;
I, pulverize, sieve: dried polysaccharide is carried out crushing screening;
It is characterized in that among the step C, the consumption of extracting solution is tenaculat Habenaria 2-3 times, is heated to and boils; Extracted 3-4 hour, among the step D, it is centrifugal that separating machine is that disc centrifuge or three whizzers cooperate tubular-bowl centrifuge to carry out, among the step E; To original volume 1/3-1/5, in the step F, ethanol concn is 95% with liquid concentration, and filling rate is 3-4 times of solution after concentrating; Among the step G, scavenging solution is 95%-absolute ethyl alcohol, and number of times is 2-3 time.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103819577A (en) * | 2014-03-24 | 2014-05-28 | 福州大学 | Method for preparing spirulina platensis polysaccharide |
| CN105707072A (en) * | 2014-12-03 | 2016-06-29 | 中国科学院大连化学物理研究所 | Spirulina polysaccharide and application thereof |
| CN108047344A (en) * | 2017-12-08 | 2018-05-18 | 江西中烟工业有限责任公司 | The preparation method of soybean spiral algae composite extract and its application on cork paper |
| CN108727510A (en) * | 2018-06-06 | 2018-11-02 | 广西大学 | The application of spirulina polysaccharide extract and preparation method thereof and enhancing immune function |
| CN111732671A (en) * | 2020-07-17 | 2020-10-02 | 王志忠 | Preparation method of spirulina polysaccharide |
| DE202022101508U1 (en) | 2022-03-22 | 2022-04-11 | Biswaranjan Acharya | Spirulina production system with intelligent circuit |
| CN115612849A (en) * | 2022-10-31 | 2023-01-17 | 南昌硬质合金有限责任公司 | Drying-free recovery method of hard alloy mixture |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108047344A (en) * | 2017-12-08 | 2018-05-18 | 江西中烟工业有限责任公司 | The preparation method of soybean spiral algae composite extract and its application on cork paper |
| CN108047344B (en) * | 2017-12-08 | 2019-12-31 | 江西中烟工业有限责任公司 | Preparation method of soybean spirulina compound extract and application of soybean spirulina compound extract in tipping paper |
| CN108727510A (en) * | 2018-06-06 | 2018-11-02 | 广西大学 | The application of spirulina polysaccharide extract and preparation method thereof and enhancing immune function |
| CN111732671A (en) * | 2020-07-17 | 2020-10-02 | 王志忠 | Preparation method of spirulina polysaccharide |
| DE202022101508U1 (en) | 2022-03-22 | 2022-04-11 | Biswaranjan Acharya | Spirulina production system with intelligent circuit |
| CN115612849A (en) * | 2022-10-31 | 2023-01-17 | 南昌硬质合金有限责任公司 | Drying-free recovery method of hard alloy mixture |
| CN115612849B (en) * | 2022-10-31 | 2023-11-14 | 南昌硬质合金有限责任公司 | Drying-free recovery method of hard alloy mixture |
| CN116814487A (en) * | 2023-06-26 | 2023-09-29 | 复旦大学 | Low-sugar high-protein spirulina dreg and preparation method and application thereof |
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Application publication date: 20121219 |