CN101565411A - Method for extracting fucoxanthin from brown algae - Google Patents

Abstract

A method for extracting fucoxanthin from brown algae, which is characterized in that fresh brown algae are beaten into a slurry, organic reagents are added, and antioxidants with a mass percent of 0.5-1 are added, extracted for 22-26 hours and then suction-filtered to obtain Extract the liquid, repeatedly extract until the organic layer is colorless, combine the liquids of each organic layer in a brown bottle; remove the organic solvent by rotary evaporation, add 1:1-2 volume ratio to the adherent substance obtained in the rotary evaporation vessel Ethyl acetate and water, dissolve and vibrate to separate layers, collect the organic layer, repeat 3-6 times, combine the liquids of each organic layer and concentrate by rotary evaporation to obtain the total pigment of the brown algae; use silica gel column chromatography to separate and purify to obtain high purity Fucoxanthin. The fucoxanthin prepared by the invention has high purity, simple and easy operation, and is convenient for industrial production, and provides basic conditions for further development of related industrial products and medical and health care products for the vacancy of the domestic and foreign markets for high-purity fucoxanthin.

Description

A method for extracting fucoxanthin from brown algae

technical field

The invention relates to a method for extracting fucoxanthin from brown algae

Background technique

There are about 250 genera and 1,500 species of Phaeophyta, most of which are marine except for a few genera and species that live in fresh water. Among the brown algae, there are many economic seaweeds with great economic value. Among them, kelp, wakame, hijiki, etc. are edible and medicinal. Fucoxanthin, also known as fucoxanthin or fucoxanthin, has the molecular formula C ₄₂ H ₅₈ O ₆ . It is an important carotenoid contained in brown algae, diatoms, golden algae and yellow-green algae. It has anti-obesity , anti-tumor, regulate blood sugar content, anti-angiogenesis, anti-oxidation and scavenging free radicals, inhibit lipopolysaccharide-induced inflammation, inhibit mammalian DNA replicase and other activities, and can also be used as food additives for aquaculture and poultry feeding Strengthen the disease resistance of animals, add color to egg yolk, etc. Fucoxanthin is widely distributed in brown algae.

my country is rich in brown algae resources, and the brown algae processing industry is prosperous. Fucoxanthin present in brown algae plants is a health food supplement, additive and effective anti-cancer drug, so the extraction, separation, development and utilization of fucoxanthin seems imminent.

Internationally, Japan is in the lead in the research of fucoxanthin. Domestically, it is relatively scarce. In June 2004, the Institute of Oceanology of the Chinese Academy of Sciences applied for a patent on the method of separating fucoxanthin from seaweed. This patent relates to the extraction and separation of fucoxanthin. The specific operation process is as follows: after thawing fresh seaweed or frozen seaweed at room temperature, wash with distilled water to remove surface salt ions, then remove the surface water, and then use dimethyl methylene The sulfone is leached in the dark, the leaching time is 15-60 minutes, the amount of dimethyl sulfoxide is 2-6ml/gram of seaweed, and finally the dimethyl sulfoxide is leached with a mixed solution of ethyl acetate and ammonium sulfate The pigment in the solution was extracted into ethyl acetate. The above-mentioned extraction method has the following defects: (1) The yield and purity of the fucoxanthin are relatively low. (2) The chemical reagent dimethyl sulfoxide used may cause nausea, vomiting, skin rash and the smell of garlic, onion, and oyster in the skin and exhaled air; the high volatile concentration of dimethyl sulfoxide may be inhaled by the human body. Causes headaches and dizziness; can burn and sting the skin; dimethyl sulfoxide may also penetrate the skin and, under certain conditions, introduce toxic substances into the skin. (3) The chemical reagent ammonium sulfate used is irritating to eyes, mucous membranes and skin. Thermal decomposition will produce toxic smoke; harmful combustion products are nitrogen oxides and sulfides. (4) The steps of the silica gel column chromatography method adopted for the separation and purification of fucoxanthin are cumbersome, time-consuming and labor-intensive.

At present, industrial fucoxanthin with extremely low purity has been sold in the domestic market, and the price of 5% fucoxanthin is between 300-450 US dollars per kilogram. The standard product of fucoxanthin is currently only known to be sold by Xibao Biological Company in Japan. The standard product of fucoxanthin derivatives is 4μg dissolved in 2.5ml ethanol solution, and the price is 7080 RMB.

Contents of the invention

The purpose of the present invention is to provide a method for extracting fucoxanthin from brown algae, which can overcome the above-mentioned shortcomings of the prior art, and can quickly and effectively extract fucoxanthin from brown algae.

The technical method of the present invention is characterized in that fresh brown algae are beaten into pulp, organic reagents are added, and an antioxidant with a mass percentage of 0.5-1 is added, and after leaching for 22-26 hours, suction filtration is obtained to obtain the leaching solution, and the leaching is repeated. Until the organic layer is colorless, combine the liquids of each organic layer in a tan bottle; remove the organic solvent by rotary evaporation, add ethyl acetate and water with a volume ratio of 1:1-2 to the adherent substance obtained in the rotary evaporation vessel, and dissolve After shaking and layering, collect the organic layer, repeat 3-6 times, combine the liquids of each organic layer and then concentrate by rotary evaporation to obtain the total pigment of the brown algae; use silica gel column chromatography to separate and purify to obtain high-purity fucoxanthin.

The fucoxanthin prepared by the invention has high purity, simple and easy operation, and is convenient for industrial production, and provides basic conditions for further development of related industrial products and medical and health care products for the vacancy of the domestic and foreign markets for high-purity fucoxanthin.

Description of drawings

Fig. 1 High-performance liquid chromatogram of high-purity fucoxanthin.

The ultraviolet/visible light absorption spectrum of Fig. 2 high-purity fucoxanthin (solvent is ethyl acetate).

Fig. 3 Electrospray mass spectrum of high-purity fucoxanthin.

Fig. 4 ¹ H NMR spectrum of high-purity fucoxanthin.

Fig. 5 ¹³ C NMR spectrum of high-purity fucoxanthin.

Detailed ways

Drain 1kg of wakame seaweed, cut it into pieces, add water and stir it into a slurry, pour it into a brown bottle, add 500ml of acetone reagent every day, and then add 1g of MgCO3 powder to prevent oxidation, leaching at low temperature in the dark, and let it stand After 24 hours, the acetone extract was filtered out with a Buchner funnel, and the extraction was repeated for 12 days until the acetone layer was colorless. Combine each acetone extract in a brown bottle, remove the acetone by rotary evaporation, then add ethyl acetate to the rotary evaporator to dissolve the adherents, transfer it into a separatory funnel, and then add distilled water of the same volume as ethyl acetate to shake Separate layers, take the ethyl acetate layer, repeat 3 times, combine the ethyl acetate layers and concentrate by rotary evaporation to obtain the total pigment of wakame. The concentrated fucoxanthin-containing ethyl acetate extract was subjected to silica gel column chromatography ( 5cm×80cm). Gradient elution was carried out with petroleum ether and ethyl acetate at a volume ratio of 1:0.43-9, and finally the column was washed with 1L of ethyl acetate. Fucoxanthin is the orange-yellow liquid that appears when eluted with petroleum ether: ethyl acetate at a volume ratio of 1:1. 0.27 g of fucoxanthin can be obtained from 1 kg of wakame, and the purity of the obtained fucoxanthin is nearly 90%.

The brown algae described in the present invention is kelp, carrageenan, sargassum, sea wormwood, kelp, undaria pinnatifida, hijiki or iron nail dish; the leaching is low-temperature and light-proof leaching; the organic reagent is Acetone, 70-100% ethanol solution or methanol; the addition of 0.5-1 mass percent of antioxidants in the extraction process is MgCO3 powder or vitamin C; the extraction of fucoxanthin from brown algae The method is characterized in that the silica gel column chromatography is carried out gradient elution with petroleum ether and ethyl acetate with a volume ratio of 1:0.33-0.43→1:7.3-9; the purity of the high-purity fucoxanthin It is above 88%.

Claims (7)

1, a kind of by the method for extracting fucoxanthin in the brown alga, it is characterized in that fresh brown alga making beating, add organic reagent, adding quality thousand marks again is the antioxidant of 0.5-1, suction filtration after lixiviate 22-26 hour, obtain vat liquor, lixiviate is colourless until organic layer repeatedly, merges each time organic layer liquid in the brown bottle; Rotary evaporation is removed organic solvent, adds ethyl acetate and the water of volume ratio 1: 1-2 in the rotary evaporation vessel in the adherent thing of gained, the vibration layering of dissolving back, collected organic layer, 3-6 time repeatedly, merge that rotary evaporation concentrates behind each organic layer liquid, total pigment that must this brown alga; Utilize silica gel column chromatography separating purification, get the high purity fucoxanthin.

2, as claimed in claim 1 by the method for extracting fucoxanthin in the brown alga, it is characterized in that described brown alga is sea-tangle, siliquosa Pelvetia, sargassun, Sargassum, kelp, wakame, Sargassum fusiforme or okamura ishige algae.

3, as claimed in claim 1 by the method for extracting fucoxanthin in the brown alga, it is characterized in that described lixiviate is the low-temperature dark lixiviate.

4, as claimed in claim 1 by the method for extracting fucoxanthin in the brown alga, it is characterized in that described organic reagent is aqueous ethanolic solution or the methyl alcohol of acetone, 70-99%.

5, as claimed in claim 1 by the method for extracting fucoxanthin in the brown alga, the antioxidant that it is characterized in that adding in the described leaching process quality thousand marks and be 0.5-1 is MgCO3 powder or vitamins C.

6, as claimed in claim 1 by the method for extracting fucoxanthin in the brown alga, it is characterized in that described silica gel column chromatography is to carry out gradient elution with sherwood oil and the ethyl acetate of volume ratio 1: 0.33-0.43 → 1: 7.3-9.

7, as claimed in claim 1 by the method for extracting fucoxanthin in the brown alga, it is characterized in that described high purity fucoxanthin purity is more than 88%.

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