CN103833692B - Method for extracting high-purity fucoxanthin from seaweeds - Google Patents

Abstract

The invention discloses a method for extracting high-purity fucoxanthin from seaweed, which is characterized in that it comprises the steps of washing the seaweed with distilled water to remove surface salt ions, and then removing the surface moisture; After 8-24 hours of leaching, suction filtration is obtained to obtain the extract solution; repeated extraction until the extract solution is colorless, and the step of combining each extraction solution in a brown bottle; finally, after a period of time of rotary evaporation, black and green appear in the solution Precipitate, filter, and remove pigment impurities; continue to rotate and evaporate until a red solid is precipitated in the solution, filter, and the obtained red solid is the product. The advantage is that it is the first time to obtain a safe, healthy, and high-purity product without using any chromatographic technology for separation. The fucoxanthin raw material is safe, low-toxic, economical and environmentally friendly during the preparation process.

Description

A method for extracting high-purity fucoxanthin from seaweed

technical field

The invention relates to a method for extracting fucoxanthin, in particular to a method for extracting high-purity fucoxanthin from seaweed.

Background technique

Fucoxanthin (fucoxanthin), its molecular formula is:

                                                 

Also known as fucusanthin, its crystal or solution is red under visible light. It is one of the important members of the carotenoid family. As an active secondary metabolite of seaweed, it is widely distributed in the marine environment and is a brown algae in algae. The main carotenoid components of seaweeds of the class Diatomaceae, Chrysophyceae, and Dinophyceae, especially the characteristic carotenoid components of seaweeds of the class Phaeophyceae. Fucoxanthin is the first molecule isolated from brown algae with a special allene carbon skeleton. It has a strong antioxidant capacity and is a strong chain-cutting antioxidant. This antioxidant effect can effectively Eliminate excess free radicals and singlet oxygen in the human body to keep the body in a healthy state, thereby effectively preventing free radical damage caused by ultraviolet rays, diabetes and Alzheimer's syndrome (AD) and other diseases. Modern pharmacological studies have shown that fucoxanthin has various activities such as anti-tumor, weight loss, anti-oxidation, anti-inflammation, anti-diabetes, and anti-angiogenesis. At the same time, biotoxicity experiments show that fucoxanthin has the characteristics of small toxic and side effects and does not affect the lipid metabolism cycle, so fucoxanthin is a marine active natural product with great potential medicinal value and economic value.

Fucoxanthin is a natural active substance derived from marine algae. Because it has strong antioxidant activity and is a provitamin A compound, it has a variety of very important potential development and utilization values. It is a new marine functional product worth developing. It can be used as: (1) drugs, such as anti-tumor, anti-aging drugs, ophthalmic drugs, cosmetic additives, etc.; (2) food and beverage additives; (3) aquatic product bait, improve immunity and increase color; (4) Feed for poultry and livestock, improve immunity, deepen the color of egg yolk, and increase the nutritional value of poultry eggs. At present, cosmetics and fucoxanthin soft capsules with fucoxanthin as the main component have been launched in the world. Among them, fucoxanthin soft capsules have anti-tumor, diabetes treatment, weight loss and other effects, and have won good reputation and comparison. High market share.

At present, the extraction and purification process of high-purity fucoxanthin mainly relies on chromatographic technology, such as a method for separating fucoxanthin from seaweed (public number CN 1706836 A), and a method for purifying fucoxanthin The method (publication number is CN 101735178A), the name is a method for enzymatically preparing fucoidan and fucoxanthin (publication number is CN 103232552 A), in the above patent, the purification process of high-purity fucoxanthin Both adopt chromatographic methods, not only the purification cost is high, but also the environmental pollution in the purification project is serious. Therefore, there is an urgent need for an extraction and purification technology with simple extraction process, no pollution, and suitable for industrial production, so as to meet the demand for high-purity fucoidan in domestic and foreign markets. Flavin requirements.

Contents of the invention

The technical problem to be solved by the present invention is to provide a method for extracting high-purity fucoxanthin from seaweed, which is simple and fast and does not require any chromatographic means.

The technical scheme adopted by the present invention to solve the above-mentioned technical problems is: a method for extracting high-purity fucoxanthin from seaweed, specifically comprising the following steps:

(1) Thaw fresh seaweed or frozen preserved seaweed at room temperature, wash 3-5 times with seawater to remove surface impurities, then wash 2-3 times with distilled water to remove salt ions on the surface of algae, and then remove its surface moisture ;

(2) Add the organic reagent to the seaweed, extract in the dark for 8-24 hours, and then filter with suction to obtain the extract;

(3) Repeat the above step (2) to extract the seaweed repeatedly until the extract is colorless; fucoxanthin is a fat-soluble substance that can be dissolved in organic reagents, and the fucoxanthin is completely extracted by organic solvent extraction. Soluble in organic solvents;

(4) Combine the above-mentioned extracts into a brown bottle, and under the condition of 15-40°C, vacuum rotary evaporation until a large amount of black and green precipitates gradually appear in the solution, and filter to remove the black and green precipitates; the temperature during the concentration and extraction process is The key influencing factor is that fucoxanthin is easy to decompose when exposed to light, so extraction and concentration cannot be carried out under high temperature conditions. Fucoxanthin is easy to decompose when the temperature is higher than 40°C, and the temperature is lower than 15°C, which is not conducive to the leaching of fucoxanthin by rotary evaporation. carried out;

(5) Take the filtrate at 15-40°C, vacuum concentrate and filter to remove the black-green precipitate;

(6) Repeat the operation of the above step (5) until a red solid is precipitated in the solution, and the red solid is collected by filtration to obtain high-purity fucoxanthin with a purity of more than 85%.

The frozen-preserved seaweed described in step (1) is obtained by washing fresh seaweed with seawater at -10°C to -5°C and freezing it for 2-8 hours.

The seaweeds described in the step (1) are copper algae, sargassum, hijiki, kelp, sargassum, fucus, chickweed, sea wormwood, kelp, wakame, kelp, macroalgae, cyst Algae, Isochrysis, Chrysalis, Phaeodactylum tricornutum, Chaetoceros gracilis, or Nitzches.

The organic reagent described in the step (2) is acetone, methanol or 60-99% (V/V) aqueous ethanol.

The ratio of the seaweed in step (2) to the organic reagent is 200g: (600-800)ml.

Compared with the prior art, the present invention has the advantages that: a method for extracting high-purity fucoxanthin from seaweed in the present invention, firstly, the fucoxanthin is completely dissolved in the organic solvent through leaching with an organic solvent to obtain The organic solvent extract contains not only fucoxanthin, but also a variety of other fat-soluble components, such as carotenoids, lipids, chlorophylls, steroids, etc. The above fat-soluble substances have the same effect in the same organic solvent Different solubility, use the obvious difference in solubility to achieve the purpose of selective extraction of fucoxanthin, so that fucoxanthin can be effectively separated from other fat-soluble components; when the extract is concentrated by rotary evaporation, carefully observe the change of the solution, gradually Concentrate to make the fat-soluble impurities precipitate first. When the extract is concentrated to a certain extent, the fat-soluble impurities are basically precipitated completely, but the fucoxanthin has not yet precipitated; continue to concentrate, and the red solid precipitated in the extract is high-purity fucoxanthin white. The advantages are as follows:

1. Simple operation. The operation process of the present invention is very simple, the separation effect is good, and the extraction reagents and instruments and equipment used are very few, so the cost is low and the cost is low;

2. Fast separation speed. The present invention does not use any chromatographic separation technology. During the process of rotary evaporating the leaching solution, the difference in solubility of different chemical components in seaweed in organic solvents is utilized, and the specific gravity of the organic solvent is gradually reduced, so that different chemical components are separated in the solution step by step. , to achieve the goal of separating and obtaining high-purity fucoxanthin;

3. The product is safe, healthy and of high purity. The invention adopts common solvents to extract fucoxanthin from seaweed, and the reagent methanol, ethanol or acetone used will not affect the chemical structure of natural fucoxanthin. Through the operation process of the present invention, safe, healthy and high-purity fucoxanthin with a purity greater than 85% can be obtained;

4. Lay the foundation for factory production. The invention proposes a simple and efficient fucoxanthin extraction method, and the extraction process is suitable for large-scale industrial production.

In summary, the present invention proposes for the first time a method for extracting high-purity fucoxanthin without chromatographic methods, which is a safe, healthy and low-cost method for preparing high-purity fucoxanthin. The invention provides a safe and low-toxicity extraction and separation technology to meet domestic and foreign high-purity fucoxanthin markets, greatly improves the added value of seaweed products, and has important economic value.

Description of drawings

Fig. 1 is the process flow chart of the present invention extracting high-purity fucoxanthin from seaweed;

Fig. 2 is the ultraviolet/visible absorption spectrogram of the fucoxanthin that embodiment 1 extracts from copper algae;

Fig. 3 is the high performance liquid phase chromatogram of the fucoxanthin that embodiment 1 extracts from copper algae;

Fig. 4 is the ultraviolet/visible absorption spectrogram of the fucoxanthin that embodiment 2 extracts from Sargassum;

Figure 5 is a high performance liquid chromatogram of fucoxanthin extracted from Sargassum in Example 2.

Detailed ways

The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

1. Experimental method

Liquid chromatography conditions:

Instruments used: Waters 2695 high performance liquid chromatography

Liquid phase conditions: stationary phase is YMC-Pack ODS-A column (250×4.6mm, 5μm); column temperature: 25 ℃; flow rate: 1 mL/min; detection wavelength: 450 nm; injection volume: 10 mL; Phase: A: methanol, B: water, gradient elution: phase A (volume %), 0-40 min (20-90), 40-50 min (90).

2. Specific examples

A method for extracting high-purity fucoxanthin from seaweed, specifically comprising the following steps:

(1) Thaw fresh seaweed or frozen preserved seaweed at room temperature, wash 3-5 times with seawater to remove surface impurities, then wash 2-3 times with distilled water to remove salt ions on the surface of algae, and then remove its surface moisture ;

(2) Add the organic reagent to the seaweed, extract in the dark for 8-24 hours, and then filter with suction to obtain the extract;

(3) Repeat the above step (2) to extract the seaweed repeatedly until the extract is colorless;

(4) Combine the above extracts into a brown bottle, and vacuum rotary evaporation at 15-40°C until a large amount of black-green precipitates gradually appear in the solution, and filter to remove the black-green precipitates;

(5) Take the filtrate at 15-40°C, vacuum concentrate and filter to remove the black-green precipitate;

(6) Repeat the operation of the above step (5) until a red solid is precipitated in the solution, and the red solid is collected by filtration to obtain high-purity fucoxanthin with a purity of more than 85%.

The frozen preserved seaweed in the above step (1) is obtained by washing the fresh seaweed with seawater and freezing it at -10 to -5°C for 2-8 hours.

The above-mentioned seaweeds are copper algae, sargassum, hijiki, kelp, sargassum, fucus, chickweed, sea wormwood, kelp, wakame, heckled seaweed, macroalgae, cyst algae, isochrysis, Chrysopsis dinoflagellates, Phaeodactylum tricornutum, Chaetoceros slenderis, Nitzches.

The above organic reagent is acetone, methanol or 60-99% (V/V) ethanol aqueous solution; the ratio of seaweed to organic reagent is 200g: (600-800) ml.

The natural fucoxanthin obtained by the present invention has the characteristics of high activity, safety and health, and can be used in the research and development of life and health products such as anti-oxidation, anti-tumor, weight loss, vision protection, diabetes medicine, cosmetic additives, etc., and can also be used in the development of food and beverages , Aquatic product bait and livestock feed.

The preparation scheme adopted in the present invention has the characteristics of safety, low toxicity, economy and environmental protection. The product obtained by adopting the invention can be applied to the development of life and health products and the addition of food, beverage, aquatic product bait and fowl feed. When the target product is obtained by adopting the invention, the production cost is reduced, the edible safety of the product is improved, and the environmental pollution is reduced.

Example 1

Wash fresh copper algae 3 times with seawater to remove surface impurities, then wash 2 times with distilled water to remove salt ions on the surface of the algae, and then use absorbent cotton to dry the surface water. Weigh 200g of copper algae into a 1000mL conical beaker, add 600mL of 80% (V/V) ethanol aqueous solution, seal the mouth of the conical beaker with two layers of safety film, extract in the dark for 12 hours, and filter. Continue to add 600mL of 80% (V/V) ethanol aqueous solution to the copper algae that has undergone the above extraction treatment, seal the mouth of the triangular beaker with two layers of safety film, extract in the dark for 12 hours, and filter. The copper algae was repeatedly extracted in the dark until the extract was colorless;

Combine the above extracts and pour them into a brown bottle, and evaporate in vacuum at 40°C. Pay attention to the changes of the solution when concentrating. When a large amount of black and green precipitates gradually appear in the solution, filter to remove pigment impurities, that is, black and green precipitates; Take the filtrate and continue vacuum concentration at 40°C until a black-green precipitate appears in the solution, and filter; take the filtrate and repeatedly concentrate and filter to remove the black-green precipitate until a red solid is precipitated in the solution (measure the specific gravity of ethanol in the solution with an alcohol meter to be 33% (V /V)), the obtained red solid is high-purity fucoxanthin with a purity of more than 85%.

The copper algae preserved in the freezer is obtained by washing fresh copper algae with sea water and then freezing them at -10 to -5° C. for 2-8 hours.

The obtained high-purity fucoxanthin was scanned at a wavelength of 200-700 nm by using an ultraviolet-visible spectrophotometer, and methanol was used as a blank reference. The results show that fucoxanthin methanol solution has the largest absorption peak at 451.4nm, which is basically consistent with the reports of fucoxanthin literature, indicating that the extracted substance is fucoxanthin, and the ultraviolet absorption spectrum of fucoxanthin is shown in Figure 2 . The HPLC system was used to analyze the obtained high-purity fucoxanthin. The specific liquid chromatogram is shown in Figure 3. According to the self-generated report of the HPLC system, the purity of the obtained fucoxanthin can reach 90.1%.

Example 2

Thaw frozen sargassum at room temperature, wash 5 times with seawater to remove surface impurities, and then wash 3 times with distilled water to remove salt ions on the surface of algae, and then dry the surface with absorbent paper. Weigh 200g of Sargassum and put it into a 1000mL beaker, add 700mL of 75% (V/V) ethanol aqueous solution, seal the mouth of the beaker with two layers of safety film, leaching in the dark for 8 hours, filter, and continue to add Sargassum Add 600mL of 80% (V/V) ethanol water solution again, seal the mouth of the triangular beaker with two layers of safety film, extract in the dark for 12 hours, and filter. The sargassum was repeatedly extracted in the dark until the extract was colorless;

Combine the above extracts and pour them into a brown bottle, and vacuum rotary evaporation at 38°C. Pay attention to the changes of the solution when concentrating. When a large amount of black and green precipitates gradually appear in the solution, filter to remove pigment impurities, that is, black and green precipitates; Take the filtrate and continue to concentrate in vacuo at 38°C until black-green precipitates appear again in the solution, and filter; take the filtrate to repeatedly concentrate and filter to remove the black-green precipitates until red solids are precipitated in the solution, and the specific gravity of ethanol in the solution is measured with an alcohol meter to be 28% ( V/V), the obtained red solid is high-purity fucoxanthin with a purity of more than 85%.

The obtained high-purity fucoxanthin was scanned at a wavelength of 200-700 nm by using an ultraviolet-visible spectrophotometer, and methanol was used as a blank reference. The results show that the fucoxanthin methanol solution has the largest absorption peak at 450.2nm, which is basically consistent with the fucoxanthin literature reports, indicating that the extracted substance is fucoxanthin, and the fucoxanthin ultraviolet absorption spectrum is shown in Figure 4 . The HPLC system was used to analyze the obtained high-purity fucoxanthin. The specific liquid chromatogram is shown in Figure 5. According to the self-generated report of the HPLC system, the purity of the obtained fucoxanthin can reach 86.5%.

Example 3

The same as the above-mentioned Example 1, the difference is that: copper algae dark leaching time is 24 hours; vacuum rotary evaporation temperature is 15 ° C; organic reagent is acetone; the ratio of copper algae to organic reagent is 200g:800ml.

Example 4

The same as the above-mentioned Example 1, the difference is that: copper algae dark leaching time is 20 hours; vacuum rotary evaporation temperature is 20 ° C; organic reagent is methanol; the ratio of copper algae to organic reagent is 200g: 650ml.

Example 5

With above-mentioned embodiment 1, its difference is: organic reagent is the aqueous ethanol solution of 60% (V/V); The ratio of copper algae and organic reagent is 200g: 750ml.

Example 6

With above-mentioned embodiment 1, its difference is: organic reagent is the aqueous ethanol solution of 99% (V/V); The ratio of copper algae and organic reagent is 200g: 750ml.

In addition to copper algae and sargassum, the above-mentioned seaweeds also include hijiki, kelp, sargassum, fucus, chickweed, sea wormwood, kelp, wakame, henna, macroalgae, cystic algae, Chrysophytes, Chrysopsis dinoflagellates, Phaeodactylum tricornutum, Chaetoceros teniosa, and Nitzches are not listed one by one here.

Of course, the above descriptions are not intended to limit the present invention, and the present invention is not limited to the above examples. Changes, modifications, additions or substitutions made by those skilled in the art within the essential scope of the present invention shall also belong to the protection scope of the present invention.

Claims (4)

1. from marine alga, extract a method for high purity fucoxanthin, it is characterized in that specifically comprising the following steps:

(1) marine alga of fresh marine alga or stored frozen is at room temperature thawed, with seawater cleaning 3-5 time, remove surface impurity, then with distilled water cleaning 2-3 time, remove the salt ion on frond surface, then remove its surface-moisture;

(2) join in marine alga by organic reagent, lucifuge lixiviate is suction filtration after 8-24 hour, obtains vat liquor; Wherein said organic reagent is the aqueous ethanolic solution of 60-99 volume %;

(3) repeat above-mentioned steps (2) operation, by marine alga repeatedly lixiviate until vat liquor is colourless;

(4) be incorporated in brown bottle by above-mentioned each vat liquor, under the condition of 15-40 DEG C, rotary evaporation in vacuo, until progressively occur in solution that a large amount of blackish green precipitates, is crossed and is filtered blackish green precipitation;

(5) get filtrate under the condition of 15-40 DEG C, vacuum concentration is crossed and is filtered blackish green precipitation;

(6) repeat above-mentioned steps (5) operation, until separate out red solid in solution, cross leaching red solid, namely obtain the high purity fucoxanthin that purity is more than 85%.

2. a kind of method extracting high purity fucoxanthin from marine alga according to claim 1, it is characterized in that: the marine alga of the stored frozen described in step (1) is by fresh marine alga, in-10 DEG C ~-5 DEG C after cleaning with seawater, within freezing 2-8 hour, obtain.

3. a kind of method extracting high purity fucoxanthin from marine alga according to claim 1, is characterized in that: the marine alga described in step (1) is Sargassum horneri, sargassum thunbergii, Sargassum fusiforme, sea-tangle, sargassun, black wrack, chickweed, Sargassum, kelp, wakame, okamura ishige algae, bulk kelp, capsule algae, Isochrysis galbana, determine whip chrysophyceae, Phaeodactylum tricornutum, Chaetoceros gracilis, rhombus algae.

4. a kind of method extracting high purity fucoxanthin from marine alga according to claim 1, is characterized in that: the ratio of the marine alga described in step (2) and described organic reagent is 200g:(600-800) ml.