CN1559647A - Preparation method of seaweed antioxidation active component - Google Patents
Preparation method of seaweed antioxidation active component Download PDFInfo
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- CN1559647A CN1559647A CNA2004100142761A CN200410014276A CN1559647A CN 1559647 A CN1559647 A CN 1559647A CN A2004100142761 A CNA2004100142761 A CN A2004100142761A CN 200410014276 A CN200410014276 A CN 200410014276A CN 1559647 A CN1559647 A CN 1559647A
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Abstract
A process for preparing the active antioxidizing component from seaweeds features that the active antioxidizing component is extracted from chlorella by supercritical CO2 at 32-47 deg.C under 35-40 MPa for 1.5-2.5 hr. Its produce can be used for improving immunity, delaying sanility and preventing cardiovascular and cerebrovascular diseases.
Description
One technical field
The preparation method of a kind of marine alga oxidation-resistant active ingredient of the present invention belongs to marine biotechnology, is used to develop the base-material of Medicines and Health Product and functional food.
Two technical backgrounds
Marine bioactivity structure of matter novelty, function uniqueness are the fields of enlivening of world today's scientific research of new drug research exploitation and the exploitation of modern medicine healthcare products.
From many achievements that the research to the marine algae biologically active substance obtains, marine drug is realizing that this great target has important potentiality and advantage.Utilize the unique functional food of marine bioactivity material exploitation, these are indiscoverable in the terrestrial organism body, and as the EPA in the ocean, DHA just seldom has in terrestrial organism.Various immunological diseases such as current cancer, cardiovascular diseases, acquired immune deficiency syndrome (AIDS) and epidemic disease constitute very big threat to human existence, the effect anticancer, antiviral, antimycotic and immunoregulation effect of existing Lu Sheng natural drug and chemical synthetic drug is undesirable, and side effect is a lot, even causes death.The ocean is a treasure-house of containing many pharmacological active substances.Develop effective healthcare products and functional food, can reduce the huge medical expenses of society, can pass through the food preventing disease.
Contain abundant antioxidant component in the marine organisms, significant antioxidant role is arranged.
In the marine bioactivity material research process, separation and Extraction is one of step of most critical.Marine organisms are of a great variety, but bioactive substance content is humble, and extractive techniques such as classical precipitation, crystallization, absorption, extraction, ion exchange technique exist bigger difficulty and limitation for the extraction and the purifying of marine active substance.Shortcomings such as the method for organic solvent extraction is mostly adopted in the extraction of marine bioactivity material, exists difficult solvent recovery, and operation is loaded down with trivial details, and environmental pollution is serious.
The supercritical liq technology successfully has been used for marine bioactivity material such as separation and Extraction EPA and DHA from marine organisms.This method is compared with sepn process routinely, have simple to operate, the extraction efficiency height, no noxious solvent is residual in the product, there is not waste discharge can not cause environmental pollution, usually use carbonic acid gas to make extraction agent in the extraction, inexpensive, be easy to get, nontoxic, safety can also recycle, and is particularly suitable for separating the material of thermo-sensitivity and easy oxidation.
Modern separation technology such as supercritical liquid extraction technique have important application prospects in extraction, separation and the purifying of marine active substance.Supercritical fluid technology is when utilizing the temperature of supercutical fluid when surpassing the gas-liquid coexisting state, pressure, dissolve certain solid, liquid or some component in them, and utilize this ability from raw material, to extract useful component or remove deleterious composition, separate the purpose of purifying thereby reach.Supercritical fluid technology can optionally extract from crude substance, separation and the required component of purifying, CO
2, not nontoxic, pollution-free with physiologically active substance generation chemical reaction, be easy to from product, remove, and can recycle.
Utilize marine animals such as oyster, marine alga, sea cucumber, sea urchin, starfish, mussel, hippocampus to develop at present and had reducing cholesterol content, anti-cancer, anticancer and raise immunity, improve functional food such as functions of intestines and stomach, Antiradiation injury, research level all only rests on the s-generation functional food, lack to implement international standard, the clear and definite and quantification of the functional factor of functional food, the mechanism of action is third generation functional food clearly.Though the research to the marine bioactivity material has dropped into lot of manpower and material resources both at home and abroad, but really reach aspect the marine drug exploitation clinical application and industrialization but seldom, can obtain the very few especially of a kind new medicine certificate, mostly concentrate on anticancer aspect the research and development of marine health product and functional food and the control treating cardiac and cerebral vascular diseases.
Three summary of the invention
Technical problem:
The preparation method who the purpose of this invention is to provide a kind of marine alga oxidation-resistant active ingredient extracts high purity marine alga oxidation-resistant active ingredient.Extract yield and reach more than 7%, the marine alga oxidation-resistant active ingredient that is extracted is removed the ability height of free radical, and it is active strong to suppress linoleic acid peroxidation.Method is easy, and is with low cost, is easy to get, nontoxic, safety.
Technical scheme:
1, a kind of preparation method of marine alga oxidation-resistant active ingredient is characterized in that:
(1) 100 gram Chlorella pyrenoidesa powder are packed into supercritical CO
2Extractor has sealed extraction kettle;
(2) set the supercritical CO 2 extraction technology parameter: pressure 35-40MPa, temperature 32-47 ℃, entrainment agent consumption are 1.0-1.5ml ethanol/g material, CO
2Flow is that 20-30L/h, extraction time are 1.5-2.5h;
(3) reach the extraction time of setting after, extract is collected, merge the extract in first separating still and second still;
(4) the vacuum concentration extract gets oil-like extracts or lyophilize gets Powdered extract.
Beneficial effect
1, the raw materials used Chlorella pyrenoidesa of the present invention is a kind of, with low cost of marine alga medium green algae, is easy to get, and is nontoxic, safety.In the biology classification, Chlorella pyrenoidesa (Chlorella pyrenoidosa) belongs to Eucaryotae (Eukaryota) Chlorophyta (Chlorophyta) Chlorophyceae (Chlorophyceae) Chlorococcale (Chlorococcales) egg capsule algae section (Oocystaceae) Chlorella (Chlorella).Rise as far back as the sixties in last century, the research of aspects such as the culture condition, nutritive ingredient, physiologically active substance, food applications, waste water (fermentative production waste water, Wastewater from Pig Farm, municipal effluent) processing of chlorella has all been carried out in countries and regions such as Japan, the U.S., USSR (Union of Soviet Socialist Republics), Germany, Czech, India, Egypt and China; Chlorella has been classified as the 21st century mankind's green nourishing source heath food by FAO.
Chlorella major physiological function has: improve immunizing power and body defence capability, can suppress cancer of the stomach excision patient's leukopenia, appetite stimulator, long-term ocean navigator's flu morbidity is descended; Improve anaemia, anaemia pregnant woman's oxyphorase value and serum levels of iron are increased, anaemia women's oxyphorase value is improved, the low hemochrome anaemia patient's who causes because of chemotherapy erythrocyte number and oxyphorase value are risen; Promote intestinal peristalsis, alleviate constipation; Show that through animal experiment oral chlorella dry powder can suppress increased blood pressure, suppress the minimizing of Aorta elastin matter, the prevention of brain apoplexy also plays health-care effect to the diabetic subject, can also promote the healing of skin wound; Chlorella powder can suppress the absorption of cholesterol and promote the drainage of cholesterol, and the arteriosclerosis that inhibition is caused by food reduces hypercholesterolemia patient's serum cholesterol value; Chlorella hot water extractive substance can alleviate GOT (the Glutamic Oxalocacetic Transaminase enzyme of light disease chronic hepatitis patient, glutamic oxaloacetic transaminase (GOT)) and the activity of GPT (Glutamic Pyruvic Transaminase enzyme, glutamic-pyruvic transaminase (GPT)); Chlorella tool antioxygenation, the hot water extract adopts the ferric thiocyanide thin-layer chromatographic analysis, confirms that containing two kinds has the very component of high anti-oxidation active substance.
2, the inventive method is easy, the extraction efficiency height.Supercritical CO
2Extraction process is that utilization is in emergent pressure 7.39MPa and critical temperature has the CO of dissolving power by force more than 31.06 ℃
2Fluid extracts the chemical separating new technology of low-pole in the raw material or apolar substance, and the carbonic acid gas that utilizes in the sepn process is extraction solvent, its have nontoxic, inertia, noresidue, can recycle etc. advantage; And this method extraction efficiency height, process are easy to regulate.Supercritical CO
2Extraction has more characteristic to the extraction of heat-sensitive substance and involatile substance with separating.Supercritical CO
2Have gas and characteristics of liquids concurrently, the dissolving power of existing liquid, good the flowing and the transmission performance of gas arranged again, and along with its dissolving power of variation of associated extraction factors such as pressure and temperature has remarkable change, the optimal processing parameter of extraction can be sought out with this, and separation, the purifying process parameter of crude extract can be studied.The optimal processing parameter that the inventive method provides is easy, extracts yield and reaches 7%.
3, Seaweed Extract anti-oxidant activity height of the present invention.Determine to optimize optimal processing parameter by orthogonal test: extraction temperature, pressure, CO
2Flow, carry secretly dosage, extraction time, test (table 1) by orthogonal table L16 (45).
Table 1 L
16(4
5) the solid element-parameter list of orthogonal test
Horizontal A temperature B pressure C CO
2Flow D carries dosage E extraction time secretly
(℃) (MPa) (L/h) (ml ethanol/g material) (h)
1 32 40 20 1.0 3
2 40 35 15 0 1.5
3 47 30 30 0.5 2.5
4 55 25 25 1.5 2
Horizontal extent 32-55 25-40 15-30 0-1.5 1.5-3
Selecting the crude extract yield is performance assessment criteria, to obtain the optimum extraction process parameter.The anti-oxidant activity of extract is as final performance assessment criteria.Anti-oxidant activity with DPPH (1, the bitter diazanyl of 1-phenylbenzene-2-) method and linolic acid method Detection and Extraction thing.The ability (inhibiting rate is up to 49.23-54.16%) that the marine alga oxidation-resistant active ingredient that is extracted is removed free radical is significantly higher than BHT (inhibiting rate reaches 23.99%), Trolox (inhibiting rate reaches 25.41%), alpha-tocopherol (inhibiting rate reaches 7.90%) (table 2).
Crude extract suppresses linoleic acid peroxidation ability (the 6th day, minimum oxidation share 2.0-3.6%), is significantly higher than BHT (the 6th day, 12.8%), Trolox is (the 6th day, 13.9%) alpha-tocopherol (the 6th day, 111.3%, do not had antioxidant effect) (table 3).
Four, description of drawings
Fig. 1 supercritical CO
2Extraction process flow.
Five embodiments
Embodiment 1:
(1) takes by weighing the 100 gram Chlorella pyrenoidesa powder supercritical CO of packing into
2Extractor has sealed extraction kettle;
(2) supercritical CO 2 extraction technology parameter: pressure 35MPa, 40 ℃ of temperature, entrainment agent consumption are 1.5ml ethanol/g material, CO
2Flow is 20L/h, and wherein pressure, flow reach preset value by regulating throttling valve open degree and CO 2 high pressure revolution speed;
(3) reach the extraction time 2.5h of setting after, extract is collected, merge the extract in first separating still and second still;
(4) vacuum concentration (40 ℃ of water-baths) extract gets oil-like extracts or lyophilize and gets Powdered extract and be the marine alga oxidation-resistant active ingredient.
The material that supercritical carbon dioxide extraction is collected carries out vacuum and low temperature and concentrates, and measures the weight of extract, and the extraction yield reaches 3.5%.The marine alga oxidation-resistant active ingredient that is extracted is removed the ability (inhibiting rate is up to 54.16%) of free radical, is significantly higher than BHT (inhibiting rate reaches 23.99%), Trolox (inhibiting rate reaches 25.41%), alpha-tocopherol (inhibiting rate reaches 7.90%).
Crude extract suppresses linoleic acid peroxidation ability (the 6th day, minimum oxidation share 3.6%), is significantly higher than BHT (the 6th day, 12.8%), Trolox (the 6th day, 13.9%), alpha-tocopherol (the 6th day, 111.3%, do not had antioxidant effect).
The removing free radical activity of table 2 Seaweed Extract
Sample extraction thing yield (%) inhibiting rate (%)
1 7.78 42.03±0.51
c
2 2.15 17.35±0.13
i
3 3.97 30.38±2.29
f，g
4 4.95 41.98±0.20
c
5 3.25 37.67±1.10
d
6 3.50 54.16±4.49
a
7 3.31 33.77±0.43
e，f
8 2.49 30.35±0.92
f，g
9 7.54 49.23±1.98
b
10 3.84 29.67±0.29
g
11 3.52 29.69±2.22
g
12 3.17 37.00±6.83
d，e
13 3.54 42.38±0.40
c
14 3.28 31.59±2.36
f，g
15 2.45 46.92±0.00
b
16 2.58 31.36±0.37
f，g
BHT / 23.99±1.91
h
Trolox / 25.41±0.03
h
Alpha-tocopherol/7.90 ± 0.49
j
Annotate: in this table inhibiting rate file, there were significant differences (p＜0.05) between the data of different letter marks organizes 6 and 9 inhibiting rate height, and it is strong to show that this crude extract of two groups is removed the free radical ability
The inhibition linoleic acid peroxidation activity of table 3 Seaweed Extract
Sample first day second day the 3rd day the 4th day the 5th day the 6th day
1 76.6％ 50.0％ 25.4％ 13.2％ 8.3％ 5.8％
2 80.5％ 51.6％ 24.5％ 14.4％ 8.8％ 7.4％
3 98.7％ 51.6％ 24.4％ 12.7％ 8.9％ 6.5％
4 88.5％ 30.9％ 13.0％ 8.2％ 7.4％ 7.6％
5 58.4％ 43.7％ 18.2％ 10.6％ 7.0％ 4.6％
6 59.7％ 47.9％ 15.9％ 7.2％ 6.9％ 3.6％
7 51.9％ 36.8％ 16.4％ 7.5％ 5.8％ 4.1％
8 66.2％ 41.1％ 18.9％ 10.2％ 6.9％ 5.8％
9 33.8％ 29.5％ 6.7％ 4.4％ 3.8％ 2.0％
10 88.3％ 50.5％ 17.2％ 11.4％ 8.3％ 5.8％
11 / 17.9％ 11.1％ 7.3％ 1.5％ 0.8％
12 108.3％ 6.4％ 4.7％ 4.7％ 4.7％ 4.4％
13 77.8％ 8.1％ 4.6％ 4.4％ 5.2％ 4.6％
14 102.8％ 10.4％ 5.8％ 6.5％ 6.5％ 5.9％
15 / 2.0％ 1.8％ 2.2％ 2.4％ 3.2％
16 119.4％ 11.1％ 6.5％ 6.0％ 6.9％ 6.3％
BHT 433.2％ 29.7％ 14.9％ 13.4％ 14.5％ 12.8％
Trolox 38.9％ 30.6％ 14.8％ 14.5％ 14.9％ 13.9％
Alpha-tocopherol 322.2% 35.8% 20.8% 32.5% 80.5% 111.3%
Annotate: the numerical value in the table (percentage ratio) is test index oxidation on same day share, and its method of calculation are
(add antioxidant linolic acid degree of oxidation/do not add antioxidant linolic acid degree of oxidation) * 100%, numerical value is more little, and the linolic acid oxidising process suppresses well more, and anti-oxidant activity is strong more
Embodiment 2:
(1) 100 gram Chlorella pyrenoidesa powder are packed into supercritical CO
2Extractor has sealed extraction kettle;
(2) set the supercritical CO 2 extraction technology parameter: pressure 40MPa, 47 ℃ of temperature, entrainment agent consumption are 1.5ml ethanol/g material, CO
2Flow is 30L/h, and wherein pressure, flow reach preset value by regulating throttling valve open degree and CO 2 high pressure revolution speed;
(3) reach the extraction time 1.5h of setting after, extract is collected, merge the extract in first separating still and second still;
(4) vacuum concentration (40 ℃ of water-baths) extract gets oil-like extracts or lyophilize and gets Powdered extract and be the marine alga oxidation-resistant active ingredient.
The material that supercritical carbon dioxide extraction is collected carries out vacuum and low temperature and concentrates, measure the weight of extract, the extraction yield reaches 7.54%, the marine alga oxidation-resistant active ingredient that is extracted is removed the ability (inhibiting rate is up to 49.23%) of free radical, is significantly higher than BHT (inhibiting rate reaches 23.99%), Trolox (inhibiting rate reaches 25.41%), alpha-tocopherol (inhibiting rate reaches 7.90%).
Crude extract suppresses linoleic acid peroxidation ability (the 6th day, minimum oxidation share 2.0%), is significantly higher than BHT (the 6th day, 12.8%), Trolox (the 6th day, 13.9%), alpha-tocopherol (the 6th day, 111.3%, do not had antioxidant effect).
Claims (1)
1, a kind of preparation method of marine alga oxidation-resistant active ingredient is characterized in that:
(1) 100 gram Chlorella pyrenoidesa powder are packed into supercritical CO
2Extractor has sealed extraction kettle;
(2) set the supercritical CO 2 extraction technology parameter: pressure 35-40MPa, temperature 32-47 ℃, entrainment agent consumption are 1.0-1.5ml ethanol/g material, CO
2Flow is that 20-30L/h, extraction time are 1.5-2.5h;
(3) reach the extraction time of setting after, extract is collected, merge the extract in first separating still and second still;
(4) the vacuum concentration extract gets oil-like extracts or lyophilize and gets Powdered extract and be the marine alga oxidation-resistant active ingredient.
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| CN1302013C (en) * | 2005-02-21 | 2007-02-28 | 南京农业大学 | Method for preparing active chlorella polysaccharide |
| CN100536975C (en) * | 2006-06-23 | 2009-09-09 | 厦门大学 | Supercritical carbon dioxide extraction recovery device for waste electric appliance combustion inhibitor and method thereof |
| CN103478125A (en) * | 2013-09-16 | 2014-01-01 | 河海大学 | Method for preparing linoleic acid sustained-release algal inhibiting agent |
| CN103880933A (en) * | 2014-03-06 | 2014-06-25 | 福州大学 | Antioxidative peptide |
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| CN100536975C (en) * | 2006-06-23 | 2009-09-09 | 厦门大学 | Supercritical carbon dioxide extraction recovery device for waste electric appliance combustion inhibitor and method thereof |
| CN103478125A (en) * | 2013-09-16 | 2014-01-01 | 河海大学 | Method for preparing linoleic acid sustained-release algal inhibiting agent |
| CN103478125B (en) * | 2013-09-16 | 2015-06-17 | 河海大学 | Method for preparing linoleic acid sustained-release algal inhibiting agent |
| CN103880933A (en) * | 2014-03-06 | 2014-06-25 | 福州大学 | Antioxidative peptide |
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