KR101502660B1 - A Fucoxanthin and Fucoidan Extraction Method from - Google Patents

Abstract

The present invention relates to a method for extracting fucosanthin and fucoidan from a seaweed, comprising the following steps: 1) crushing a seaweed, 2) a step of packing the crushed seaweed into a nonwoven fabric, and 2) A fourth step of extracting the extract of the third step at a temperature of 50 ° C to 60 ° C, a fifth step of extracting fucosanthin from the filtrate of the extract of the fourth step, And a sixth step of extracting fucoidan from the marine residue of the extract of the fourth step.

Description

A method of extracting fucoxanthin and fucoidan from a seaweed,

The present invention relates to a method for extracting fucosanthin and fucoidan from a seaweed, and more particularly, to a simple method for simultaneously extracting fucoidan and fucoidan from a seaweed by a single process.

The present invention is the result of the research carried out by the industry-academia joint technology development project (convention number: 00047086).

Fucoxanthin, a component of the carotenoid system present in edible seaweeds, is contained in a small amount in marine algae (about 0.1% of marine spores) and is in an unstable state, resulting in delayed commercialization. However, Recently, many researchers have been interested in factory production development. The efficacy of fucosanthin reported in the paper is very high, including anticancer effect by suicide cells, anti-obesity effect by white fat cell destruction, prevention of hypertension, anti-inflammatory effect, antioxidant effect.

Much research has been done on extraction tablets. In 1989, Haugan et al. Of Norway published a study in which pyridine was purified as a solvent by a chromatographic method. In 2005, Wang et al. Reported that dimethyl sulfoxide is more effective in extracting fucosanthin than acetone, which is the most commonly used organic solvent.

In 2008, Oryza Oil & Fat Chemical Co. announced the extraction and purification process at the factory stage. In 2008, it reported that 3 times higher purity ethanol was used and purified by silica gel column chromatography. And produced 1490 g of fucosanthin from 10 tonnes of discarded kelp.

The following is the summary of researches and presentations about fucosanthin in Korea. Acetone (methanol) was used in the seaweeds of Mokpo National University to find that the content of fucosanthin contained in the seaweed sporangia was 1.4 times higher than that of marine leaf and 2.0 times higher than that of marine stalk.

When fucoxanthin was extracted from the gut using supercritical carbon dioxide, the content of fucoxanthin was the highest at 45.8 μg per gram of the extract. When fucoxanthin was extracted and concentrated The heavy chain fatty acid which is a vegetable derived fatty acid was selected. The three pressure and temperature conditions that showed high efficiency for fucoxanthin enrichment were 40 at 9.7 MPa, 45 at 11.0 MPa, and 50 at 12.4 MPa.

In 2011, KIST reported that fucosanthin was extracted using pressurized liquid extration (PLE) method, and that fucosanthin extraction efficiency reached 95%. They modeled the fucosanthin extraction process, which was proved by experiments to be 90% ethanol solvent.

The level of domestic technology for fucosanthin is not far behind that of developed countries in the laboratory stage, but the level of industrialization technology is at an early stage. Fucosanthin is a very expensive substance because it is contained in a small amount in seaweed, it has to be extracted with an organic solvent, and it has to undergo freeze drying process due to heat and light.

An object of the present invention is to solve the above problems and to provide a method for improving economical efficiency by extracting fucoidan from a marine residue and extracting fucosanthin from a marine residue.

To achieve the above object, the present invention provides a method for extracting fucosan and fucoidan from a seaweed, comprising the steps of: pulverizing a seaweed, a second step of packing the seaweed into a nonwoven fabric, A third step of extracting fucosanthin at a temperature of 50 ° C to 60 ° C, a third step of extracting fucosanthin from the filtrate of the extract of the fourth step, And a sixth step of extracting fucoidan from the marine residue of the extract of the fourth step.

The effect of the method of extracting fucoxanthin and fucoidan from the dried legume according to the present invention is as follows.

It is possible to extract fucoidan from the remaining marine residue by extracting fucosanthin from the seaweed and extract fucoidan and fucoidan in a simple and low-cost process.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flowchart showing a process of extracting fucosanthin and fucoidan from a seaweed according to the present invention. FIG.
FIG. 2 is a comparative diagram showing the addition of ultrasonic vibrations in the process of extracting fucosanthin from a seaweed according to the present invention. FIG.
FIG. 3 is a graph showing a change in fucosanthin yield depending on the ethanol input amount in the process of extracting fucosanthin from seaweed according to the present invention. FIG.
Fig. 4 is a graph showing changes in the amount of fucosanthin extracted with time in the process of extracting fucosanthin from a seaweed according to the present invention. Fig.
FIG. 5 is a graph showing the change in the amount of fucosanthin extracted with time of heat supply in the process of extracting fucosanthin from a seaweed according to the present invention. FIG.
FIG. 6 is a graph showing changes in the amount of fucosanthin extracted with time of light in the process of extracting fucosanthin from a seaweed according to the present invention. FIG.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

First, a method for extracting fucosanthin and fucoidan from a seaweed according to the present invention comprises a first step of pulverizing a seaweed, a second step of packing the pulverized seaweed into a nonwoven fabric, a step of adding ethanol to the seaweed, A fourth step of extracting the extract of the third step at a temperature of 50 ° C to 60 ° C, a fifth step of extracting fucosanthin from the filtrate of the extract of the fourth step, And a sixth step of extracting fucoidan from the undersea residue of the four-step extract.

In the fifth step, step 5-1 of adding the same amount of the seaweed as that of the filtrate to the filtrate and extracting the extract at a second room temperature, and a step 5-1 of extracting the extract of step 5-1 at a temperature of 50 ° C to 60 ° C (5-2) a step 5-2 of mixing the undissolved residue of the extract extracted in the step 5-2 with the undissolved residue of the sixth step, and 5-5 step of extracting fucosan in a filtration solution of the extract in a vacuum concentrator, 5-5 step of concentrating the fucosylate solution by evaporating ethanol from the extract of step 5-4, 5-5 Step 5-6 of measuring the amount of the fucosylate solution and mixing the excipient 50 to 300 times the weight of the fucosylate solution with 2 to 10 times the weight of the fucosylate solution; And (5-7) lyophilizing the mixed solution of step (6).

Step 6-1 is a step 6-1 of adding water and a trace amount of organic acid to the seaweed residue of the sixth step in an amount of 5 to 15 times the weight of the marrow residue, A step 6-2 of extracting the mixed liquor at a temperature of 90 ° C to 100 ° C and a step 6-3 of adding water of 5 to 15 times the weight of the whale residue to the seaweed residue of the extract of 6-2 , A step 6-4 of extracting the mixture of the step 6-3 at a temperature of 100 ° C to 110 ° C, a filtration solution obtained by filtering the extract of the step 6-4, A step 6-5 of adding CaCl2 to the filtrate obtained by filtration of the obtained extract, and a step 6-6 of filtering the fucoidan by filtering the mixed solution of the step 6-5 and removing alginic acid .

In the sixth to sixth steps, the desalination process is performed using an ultrafilter, and the cutoff is preferably 5000 to 30,000 Da.

Hereinafter, the order of the extraction process will be described in detail.

First, the method of extracting fucosanthin is to crush the dried seaweed and pack it in a nonwoven fabric. The pulverization has the effect of decreasing the amount of ethanol input by reducing the volume occupied by the seaweed at a particle size of 0.1 to 2 cm. By adding 3 ~ 7kg of crushed seaweed to the nonwoven fabric and wrapping it, the raw material is introduced into the extractor and the filtration process becomes easy.

Ethanol (90 to 99.5%) 1.5 to 5.0 times the weight of the added seaweed is added to the extractor and ethanol is circulated at room temperature. Extract the light thoroughly to prevent light from entering the entire process. After extraction at room temperature for 0.5 ~ 3 days, heat at 50 ~ 60 ℃ and extract at this temperature for 2 ~ 5 hours. The fucosylate concentration is measured during the extraction and the extraction stops when the concentration starts to decrease at the highest concentration.

After the first extraction, the same amount of seaweed is added to the extracted solution and the second extraction is performed. The remaining wakame solid is used for fucoidan extraction. The second extraction is conducted at room temperature for 0.5 ~ 3 days and then heated at 50 ~ 60 ℃ for 2 ~ 5 hours. The fucosylate concentration is measured during the extraction and the extraction stops when the concentration starts to decrease at the highest concentration. After the second extraction, the remaining wakame solid remaining after filtration is used for fucoidan extraction.

The fucosanthin extract is put into a vacuum concentrator and the ethanol is evaporated to concentrate the fucosanthin solution. In the vacuum concentration process, do not let the temperature exceed 60 ℃ and block the light so that fucosanthin is not destroyed.

Measure the amount of fucosanthin and mix with 50 ~ 300 times the weight of fucosanine (dextrin, soy protein, etc.) with water. The amount of water is 2 to 10 times that of the concentrate. The liquid mixed with the excipient is placed in a freeze dryer or vacuum dryer and dried. The temperature should not exceed 60 ° C during the entire lyophilization or vacuum drying step.

Meanwhile, the fucoidan extraction purification process is as follows. After the first secondary fucosanthin extraction, the remaining solid nonwoven fabric is collected and put into an extractor. Water is added 5 to 15 times the weight of seaweed. Add organic acids such as acetic acid, formic acid, and citric acid to 0.01-0.1% aqueous acid solution.

After the temperature of the extractor reaches 90-100 ° C by heating while circulating the water, it is extracted for 30-90 minutes. After the first fucoidan extraction, the nonwoven fabric is left in the extractor and the extract is transferred to the storage tank. Water is added 5 ~ 15 times of seaweed again. After the temperature of the extractor reaches 100 ~ 110 ℃ by heating with circulating water, it is secondly extracted for 30 ~ 90 minutes.

After the extraction of the second fucoidan, the nonwoven fabric is left in the extractor and the extract is transferred to the storage tank. First, measure the sugar content of the combined solutions of the primary and secondary filtrates, and add 20 to 100% of CaCl _2, which is the product of the weight and the sugar content, to make alginic acid to be calcium alginate and to be separated from fucoidan.

After removing the calcium alginate by using a filter press, the solution is put into an ultrafilter and adjusted by desalting and fucoidan molecular weight. The desalination process is a process of removing salts such as NaCl and CaCl ₂ using an ultrafiltration membrane with a cutoff of 5,000 ~ 30,000 Da.

In order to control the molecular weight of the desalted fucoidan solution, the ultrafiltration is carried out again with a membrane having a larger cutoff than that of desalting. For example, to divide the molecular weight of fucoidan liquid by 100,000, two types of fucoidan liquids can be obtained. Ultrafiltration with an ultrafiltration membrane with a cutoff of 100,000 gives a fucoidan liquid of 100,000 Da or less and a fucoidan liquid of 100,000 Da or more.

In order to increase the concentration of fucoidan liquid, it can be made into fucoidan liquid of 5 ~ 15Bx by vacuum concentration at 55 ~ 65 ℃. Or the concentrated fucoidan liquid can be evaporated in a vacuum evaporator or a freeze dryer to produce a fucoidan powder.

[Example 1] Comparison of extraction method of fucosanthin

The fucoxanthin extraction method was compared and compared. Seaweed samples were soaked in a soaking nonwoven fabric and placed in a soaking in fabric nonwoven fabric. Ultrasonic in-fabric was used to add 3 times the weight of the seaweed ethanol, and then extracted at room temperature for 2 hours 2). Ultrasonic vibration was 50kHz and fucosanthin content was analyzed by UV peak wavelength at 454nm wavelength. The extraction efficiency was 31% higher than that of ultrasonic wave (0.2406 / g) when ultrasonic wave was used and 0.3156 / g ultrasonic wave was used to improve the process. 2). In the case of not using ultrasonic waves, the extraction yield of fucosanthin should be improved by immersion for a long time.

[Example 2] -The yield change of fucosanthin according to the amount of ethanol input

The extraction yields of fucosanthin from the seaweed samples were compared according to the amount of ethanol (a multiple of the weight of the seaweed) as a solvent. The non-woven fabric was immersed in water at room temperature for 2 hours, filtered and analyzed for fucosamine content in ethanol. As the amount of ethanol increases, the amount of extraction increases slightly.

[Example 3] - Extraction amount of fucosanthin according to extraction time

In order to lower the cost of extracting fucosanthin, a method of extending the extraction time by doubling the amount of ethanol was studied. As shown in FIG. 4, the concentration of fucosanthin was increased rapidly until 6 hours and the concentration after immersion for 24 hours was 0.3226 / g. The amount of ethanol was 2 times the amount of seaweed, and it was confirmed that it was suitable for about 6 hours by immersion extraction.

[Example 4] Heat and light stability of fucosanthin

Fucosanthin is known to be sensitive to heat and light, so be careful with heat and light during manufacturing and storage. The results of the thermal stability test are shown in Fig. It was stable for 1 day up to 60 ℃, but decreased about 30% during 1 day at 70 ℃. The fucosanthin production process was stable at temperatures below 60 ° C. During the storage period of 6 days, it was confirmed that fucosanthin was slightly reduced even at 25 ° C, and storage at a low temperature of about 4 ° C was safe.

In order to confirm the light stability of fucosanthin, a fucosanthin concentration change with time was measured with a 60 W bulb at a distance of 1 m. As shown in FIG. 6, after one day, it was decreased to 50% or less, confirming that fucosanthin was very weak to light. It has been shown that light blocking should be packed in a perfect container when stored.

[Example 5]

5kg of nonwoven fabric is crushed into an average particle size of 0.3cm, and the resulting mixture is packed into a nonwoven fabric and packed into an extractor.

The ethanol (95% purity) 2.0 times the weight of the added seaweed (50 kg) is put into the extractor and the ethanol is circulated at room temperature. Extract the light thoroughly to prevent light from entering the entire process. After 1 day of room temperature extraction, the mixture was heated to 50 ~ 60 ℃ and the concentration of fucosanthin was measured at this temperature. When it reached 3 hours, the concentration of fucosanthin became 95 ppm and the concentration was decreased to stop extraction. After the first extraction, 50 kg of the same amount of seaweed was added to the extracted solution and the second extraction was started at room temperature. The remaining seaweed solid was used for fucoidan extraction. Secondary extraction, like primary, was heated to 50 ~ 60 ℃ for 1 day and extracted for 4 hours at this temperature. The maximum concentration reached 176ppm and extraction was stopped. After the second extraction, the remaining wakame solid was filtered and used for fucoidan extraction.

The fucoidan extract was put in a vacuum concentrator and concentrated to 1/4 of the initial amount in a state of blocking light at a temperature of 45 to 49 캜. The amount of fucosanthin was measured, and soybean protein powder, 100 times the weight of fucosanthin, was added together with water and mixed. The amount of water was 6 times that of the concentrate. The solution mixed with the excipient was placed in a freeze dryer and dried. After freeze-drying, the yield of fucosanthin was 0.03%.

[Example 6]

After extracting the first secondary fucosanthin of Example 5, the nonwoven fabric containing the residual solids is collected and put into the extractor. Water was added 10 times the weight of the seaweed, and citric acid was added to make 0.03% aqueous acid solution. The mixture was heated while circulating the water and extracted at an extractor temperature of 95 to 100 ° C for 60 minutes. After the first fucoidan extraction, the nonwoven fabric was left in the extractor, and the extract was transferred to a storage tank. Water was added to the extractor 8 times the weight of the seaweed. The mixture was heated while circulating the water, and then subjected to secondary extraction at an extractor temperature of 100 to 105 ° C for 40 minutes. After the extraction of the second fucoidan, the nonwoven fabric was left in the extractor and the extract was transferred to the storage tank. After adding 40% CaCl _2, which is the product of the weight of the extract and the sugar content, stirring the solution for 2 hours to make alginic acid to be calcium alginate to be separated from fucoidan . Calcium alginate was removed using a filter press, and the solution was added to an ultrafilter to adjust the desalting and fucoidan molecular weights. NaCl and CaCl ₂ salts were removed by ultrafiltration membrane with cutoff of 10,000 Da. In order to increase the concentration of fucoidan solution, it was concentrated at a temperature of 60 to 65 ° C under vacuum to prepare a fucoidan solution having a sugar content of 6.0 Bx. As a result of calculating the weight of 6.0bx solution, the yield of fucoidan based on seaweed was 5.2%.

It is to be understood that the invention is not limited to the disclosed embodiment, but is capable of many modifications and variations within the scope of the appended claims. It is self-evident.

Claims (7)

A first step of pulverizing the seaweed;
A second step of packing the ground wax into the nonwoven fabric;
A third step of adding ethanol to the seaweed and extracting the seaweed at room temperature;
A fourth step of extracting the extract of the third step at a temperature of 50 ° C to 60 ° C for 2 to 5 hours;
A fifth step of isolating light from the filtrate of the extract of the fourth step and extracting fucosanthin at 60 ° C or lower; And,
And a sixth step of extracting fucoidan from the undersea residue of the extract of step 4, wherein the fucoidan is extracted from the seaweed residue.
In the fifth step,
A seaweed having the same mass as that of the filtrate was added to the filtrate, Step 5-1: Extraction;
A step 5-2 of extracting the extract of step 5-1 at a temperature of 50 ° C to 60 ° C;
A step (5-3) of mixing the undissolved residue of the extract extracted in the step 5-2 with the undissolved residue of the step 6;
(5-4) extracting fucosanthin from a filtrate among the extracts extracted in the step 5-3 into a vacuum concentrator;
(5-5) of evaporating ethanol from the extract of step 5-4 to concentrate the fucosylate solution by blocking light at 60 ° C;
Measuring the amount of the fuco acid-containing liquid in the step 5-5, and mixing the excipient 50 to 300 times the weight of the fuco acid-containing liquid with 2 to 10 times the weight of the liquid of the fuco acid-containing liquid; And,
And (5-7) lyophilizing the mixed solution in steps 5 and 6, wherein the fucoxanthin and fucoidan are extracted from the seaweed. delete The method according to claim 1,
In the sixth step,
Step 6-1) adding water and organic acid to the seaweed residue of step 6 in a weight of 5 to 15 times the weight of the marrow residue;
(6-2) extracting the mixed solution of step 6-1 at a temperature of 90 ° C to 100 ° C;
A step 6-3 of adding water of 5 to 15 times the weight of the marine residue to the marine residue of the extract of 6-2;
A step 6-4 of extracting the mixture of step 6-3 at a temperature of 100 ° C to 110 ° C;
A step 6-5 of adding CaCl2 to the filtrate obtained by filtering the extract of step 6-4 and the filtrate obtained by filtering the extract obtained in step 6-2; And,
And filtering the fucoidan in the filtration solution containing CaCl ₂ in step 6-5, thereby removing alginic acid, and filtering the fucoidan. The method of claim 3,
In the sixth to sixth steps,
The filtration method for removing the alginic acid and filtering the fucoidan,
Wherein the desalting process is performed using an ultrafilter, wherein the cutoff is 5000 to 30000 Da. The method of extracting fucosanthin and fucoidan from a seaweed. The method according to claim 1,
In the third step,
The method of extracting fucosanthin and fucoidan from a marine wakame is characterized in that ultrasonic vibration is further added in the process of adding and extracting ethanol. The method according to claim 1,
In the third step,
Wherein the weight of the ethanol is 1.5 to 5 times the weight of the seaweed, and the ethanol is extracted at room temperature for 0.5 to 3 days. The method for extracting fucosan and fucoidan from a seaweed. The method according to claim 1,
In the fourth step,
Wherein the extract is extracted at a temperature of 50 to 60 DEG C for 2 to 5 hours to extract fucosanthin and fucoidan from the seaweed.

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