RU2360545C1 - Kelp processing method - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: invention relates to technology of kelp processing. Raw or frozen kelp is hydrolysed by hydrochloric acid water solution, extract is drained off and put to water-soluble polysaccharides extraction by ethanol sedimentation. The rest is extracted by 96% ethanol to obtain mixture, containing end products, obtained spirit extract is concentrated. Mixture of free fatty acids, sterine, derivate colourants is extracted from concentrate by hexane solvent. After that mixture of polyphenol compounds and colourants is extracted by dichloromethane or chloroform. High purity end products are extracted on silica gel at lowered pressure with help of column chromatography. Free fatty acids, sterols, chlorophyllous and carotinoid derivates are extracted by hexane extract. Dichloromethane or chloroform extract are used for producing fine complex of polyphenol compounds. Kelp left after spirit extraction is dried on air and used for alginic acids salts producing.

EFFECT: invention allows to obtain chemically different high purity biologically active substances free fatty acids, polyphenol compounds, sterols, colourants derivates, water-soluble polysaccharides, alginic acids salts with high yields.

2 cl, 7 ex

Description

The invention relates to a technology for processing seaweed brown algae of the Laminariaceae family (family Laminariacea) to produce biologically active substances of different chemical nature, and especially free fatty acids and polyphenolic compounds, as well as phytosterols, water-soluble polysaccharides, salts of alginic acids.

The relevance of the task of obtaining the above biologically active substances is due to the following reasons.

Fatty acids, especially polyenoic (polyunsaturated, PUFA), are necessary for the development and normal functioning of the brain, visual system, and skin. The high consumption of polyene fatty acids in the diet provides a reduction in cardiovascular and inflammatory diseases, affects processes related to human growth, reproduction, innate and acquired immunity. Polyenic fatty acids have anticancer, antitumor, antimetastatic activities (Noda N., et al., Hydrobiologia, 1990, 204/205, 577-584; Vries CE de Norden CJ van. Anticancer Res., 1992, 12, 1513-1522; Sukenik A., et al., Ann. Nutr. Metab., 1994, 38, 85-96; Roberts DD, et al., Arch. Biochem. Biophys., 1998, 267, 405-415), antithrombotic and antiarrhythmic activities (Carroll DN, et al., Ann. Pharmacotherapy, 2002, 36, 1950-1956).

Currently, preparations have been created that contain from 30% to 45% PUFA of the ω3 series: Omega-3 (USA), MaheRA (France), Eikonol (Russia) and Tecom (Ukraine). Experimental and clinical studies have revealed significant hypolipidemic, fibrinolytic, antiaggregant, hypocoagulating and immunomodulating properties in the drugs, explained by the high content of PUFAs of the ω3 series. The protective properties of drugs with respect to inflammatory and fibrotic processes in the lungs, stimulation of phagocytosis (Putintseva N.V. Ukrainian Pulmonary Journal, 2003, 4, 56-58) were established.

PUFAs are used in drugs used in the treatment of inflammation, such as rheumatoid arthritis, osteoarthritis, dementia, including Alzheimer's disease. PUFAs isolated from algae are considered promising hepatoprotectors and antioxidants. The introduction of PUFAs in the composition of drugs helps to restore liver function and increases the resistance of liver cells to the effects of damaging agents (Nazarchuk Yu.S. et al. V International scientific and practical conference of students, graduate students and young scientists. 2002, Kiev).

The optimal ratio of PUFAs in the ω3 series and ω6 series is their ratio in the range 4: 1-1: 1. Meanwhile, modern food contains an excess of polyenoic acids of the ω6, ω9 series due to the consumption of meat, mayonnaise, eggs, and vegetable oils. The lack of fatty acids of the ω3 series, which are mainly found in fish, marine invertebrates, nuts, linseed, soybean, sesame oil, is due to the poverty of our daily diet with these products. Therefore, the production of preparations containing PUFA ω3 series does not lose its relevance. Algae can be a more complete and more concentrated source of PUFA than fish oils. In this regard, kelp is of particular interest. The main fatty acids in the kelp of the Far Eastern seas are 16: 0, 18: 1 ω9, 18: 2 ω6, 18: 3 ω3, 18: 4 ω3, 20: 4 ω6, 20: 5 ω3. The proportion of PUFAs in the ω3 series ranges from 13.6 to 60.0%, the proportion of PUFAs in the ω6 series ranges from 14.5 to 28.0% of the total fatty acids, depending on the time, place of collection and age of the algae. It should be noted that seaweeds are not so rich in lipids (0.1-2.2% of wet weight, or, as several authors indicate, lipids make up 0.4-4.8% of the dry weight of algae). However, the huge reserves of algae, the possibility of their complex processing compensates for this drawback and allows us to consider algae as a potential source of lipids and fatty acids, in which PUFAs occupy a significant part.

The increased use of PUFAs by pharmaceutical companies in the formulations of various drugs and the increased demand for such drugs set the goal of increasing the production of PUFAs from alternative raw materials, including algae, in addition to fish oils and vegetable oils.

Polyphenolic compounds are no less important than PUFA organic compounds of brown algae. As shown by some authors (Ragan MA, et al., Prog. Phycol. Res, 1986, 4, 129-241; Zhang J., et al., Applied Phycology, 2006, 18, 445-450), polyphenolic compounds can be 0.3-38.0% in alcoholic extracts of brown algae. These substances act as protective agents against marine herbivores, pathogens, epiphytes, and ultraviolet radiation. Polyphenolic compounds exhibit antibacterial activity against gram-positive and gram-negative bacteria, which shows the potential for their use in the food industry and medicine. With antioxidant activity, polyphenolic compounds can replace synthetic antioxidants. Polyphenolic compounds inhibit the activity of the enzyme gualuronidase, which is included in allergic processes, the spread of cancer and inflammatory processes (Cerantola S., et al., Botanica marina, 2006, 49, 347-351). There are reports of antithrombotic and fungicidal effects of polyphenolic compounds (Method of processing brown algae. No. 2132622. 1999.07.10, A23L 1/0532, A61K 35/80, Nekrasova VB, Nikitina TV, Kurnygina VT, Belozersky O.A.).

Brown algae polysaccharides have various biologically active properties: antitumor, anticoagulant, antiviral, etc. (TNZvyagintseva et al. CBP. Part C.126. 2000, 209-215; RU 2304443 C1, 08.20.2007; RU 2247574 C1, 10.03. 2005).

Of the greatest interest are the following methods of processing algae to produce biologically active substances of various chemical nature.

A known method of producing biologically active substances from the White Sea kelp, according to which several biologically active substances are obtained from algae - mannitol, a water-soluble polysaccharide complex, sodium alginate (RU 2028153 C1, 02/09/1995). However, the known method does not allow to obtain such valuable substances as lipids, which include in their composition no less valuable fatty acids.

A known method of complex processing of brown algae, including obtaining mannitol from dried and ground algae, extracting it with 0.8-1.2% aqueous solution of mineral acid, followed by multistage purification of mannitol extract with calcium oxide or hydroxide, crystallizing it, purifying with alcohol, re-crystallizing mannitol (RU 2070808 C1, 12/27/1996). After extraction of mannitol from the residue with washed drinking water, algae are extracted with sodium alginate at pH 8.8-9.6 and calcium alginate by adding a solution containing calcium ions. The calcium alginate gel is treated with a solution of hydrochloric acid to obtain alginic acid and then its monovalent salts, calcium acid salt of alginic acid. After neutralization, the remainder of the alga is pressed, granulated and dried to be used as a feed additive.

However, the method, despite the use of multiple extraction and purification procedures, is aimed at obtaining only mannitol and salts of alginic acid, and does not affect the production of other valuable biologically active substances contained in specific types of algae, in particular lipids or fatty acids, water-soluble polysaccharides.

A known method of obtaining biologically active substances from kelp, which consists in the degreasing of algae with chloroform, followed by extraction with ethanol, hot water and a solution of sodium carbonate to obtain mannitol, the total polysaccharide-protein complex and sodium alginate (RU 2194525 C1, 12/20/2002).

However, in this method, the algae is degreased with chloroform, and valuable substances - lipids, are lost.

A known method of complex processing of brown algae to obtain iodine-containing and polysaccharide products, which provides for the extraction of crushed algae with a 65-75% ethanol solution at a temperature of 50-60 ° C for 1.5-2.5 hours, concentration of an aqueous-alcoholic extract to obtain an aqueous -lipid emulsion, which is cooled, is divided into aqueous and lipid fractions to obtain iodine-containing mineral-mannitol complex from the aqueous fraction, and iodine-containing lipid complex from the lipid fraction (RU 2233104 C1, 07.27.2004). The algal residue is used to produce fucoidan, calcium algylose and / or sodium algylose, alginic acid.

However, in this method, receiving a wide range of biologically active substances, the use of the lipid part of the algae is limited to using it as a source of iodine, and not as a source of fatty acids.

A known method of processing brown algae to obtain a fucus lipid concentrate (RU 2165720, C2, 04/27/2001) by extracting algae with an organic solvent with carbon atoms from 1 to 6. Fucus lipid concentrate contains in emulsified form water, water-soluble amino acids, proteins, polysaccharides, sugars, as well as a rich set of fatty acid esters, including ω3 type, chlorophyll derivatives, sterols, micro and macro elements. The lipid concentrate and the product made from it are saponified fucus concentrate, where fatty acids are 10-70%, pigments are 20-10000 mg%, sugars are 0.1-15%, iodine is 80-30000 μg / g, fucus sterols - 1-4%, ash substances - 3-30%, are used as dietary supplements and as part of various dietary supplements.

However, this method also deals with the process of saponification of lipids after their preliminary isolation from algae as part of a complex mixture. In the process of saponification of a lipid-containing mixture with an aqueous alkali solution, the amount of fatty acids in the form of soaps is obtained, which are used to produce a specific dietary supplement.

There is a method of complex processing of brown algae to produce individual preparations of acidic and neutral polysaccharides (laminaranes, fucoidans, polymannuronic acids) and a concentrate of low molecular weight biologically active substances recommended for use in the perfumery and cosmetic industry (RU 2240816 C1, 11.27.2004). The invention is directed to obtaining individual polysaccharide preparations with standard characteristics, the composition of the lipid part is not characterized. Mentioned only the possibility of using it in the perfumery and cosmetics industry.

There is a method of processing brown algae, namely air-dried kelp and air-dried fucus vesiculi, to obtain lipid and water-soluble concentrates from these algae (RU 2132622 C1, 10/07/1999). The lipid concentrate obtained from crushed air-dried algae by extraction with an organic solvent with a number of carbon atoms from 1 to 6 and at a boiling point of the solvent at a raw material – extractant ratio of 1: 1-1: 20 contains almost the entire lipid part of the starting algae, including ω3 series fatty acids, as well as sterols, pigments, fat-soluble vitamins, mannitol, essential amino acids and proteins, trace elements. Fucus lipid concentrate also contains polyphenols and fucoidan. Another product released from algae, a water-soluble concentrate, contains water-soluble proteins, mannitol, polysaccharides, vitamins and trace elements. As the authors indicate, lipid kelp concentrate should be used for the production of saponified kelp concentrate, which is used as a substance of the Klamin therapeutic and prophylactic additive, which has oncoprophylactic and radioprotective effects.

However, in this method, the task is to obtain a concentrate containing lipids in the complex mixture, which the authors propose to use for the production of another product - saponified kelp concentrate. It should be noted that, using the lipid saponification procedure, the authors obtain fatty acid salts (soap) in a complex mixture. However, in the production of products, including dietary supplements offered by developers, the levels of daily intake of fatty acids, and not their salts (soaps), are regulated.

As a prototype, the method of processing brown algae to obtain the drug "Mamoklam" is described in the patent (RU 2205019, C1, 05.27.2003). The tool is obtained from thalli, rhizoids and cuttings of saccharine kelp and Japanese kelp, as well as other species of the kelp family. The method includes a procedure in which lipids are first obtained by extracting them from algae with an organic solvent with the number of carbon atoms from 1 to 6. The lipid fraction - kelp concentrate - after complete distillation of the solvent is hydrolyzed with a solution of sodium hydroxide or other alkaline reagent to obtain a saponified kelp concentrate, in the composition of which includes such components as polyene fatty acids of the ω3 and ω6 series, iodine and other minerals, chlorophyll derivatives, organically bound to amino acids, policies Rida - laminaran, fucoidan. The main active ingredients in kelp concentrate and saponified kelp concentrate, as the authors indicate, are polyene fatty acids, iodine, polysaccharides laminaran and fucoidan. Both products - lipid concentrate and saponified lipid concentrate are used as a substance for the production of the drug Mamoklam.

However, in the specified method to obtain substances similar to our claimed method, the procedure for isolating complex mixtures containing lipids with organic solvents is first carried out, then the lipid mixture is hydrolyzed with a sodium hydroxide solution after distillation of the solvent to obtain fatty acids. It should be noted that when using an aqueous solution of sodium hydroxide, the authors obtain salts (soaps) of fatty acids. In the production of products, including dietary supplements, the levels of daily intake of fatty acids, and not their salts (soaps), are regulated. In addition, the known method does not provide polyphenolic compounds, phytosterols, water-soluble polysaccharides, salts of alginic acids.

All the above methods of processing algae are of undoubted interest and lead to complex mixtures, including lipids or salts (soaps) of fatty acids. These complex mixtures showed a positive effect when used as part of specific preparations. However, for the industrial production of various preparations based on biologically active substances of algae, products with standard characteristics are required to ensure the dosage of substances in the composition of the preparations in accordance with the daily intake rate.

The technical result provided by the claimed method is to expand the spectrum of biologically active substances obtained from marine brown algae.

The method is aimed primarily at isolating the sum of free fatty acids, including polyenoic ones, without first extracting lipids from algae and, in addition, allows to obtain highly purified polyphenolic compounds, phytosterols, water-soluble polysaccharides and salts of alginic acids with high yields.

Thus, we can talk about a fairly complete processing of algae to produce substances of various chemical nature.

The technical result is achieved in that the hydrolysis of lipids (storage lipids and polar lipids of biomembranes - glyceroglycolipids and phospholipids) is carried out 0.6-0.8 N. an aqueous solution of hydrochloric acid when heated within 45-50 ° C for 4-5 hours and then without heating for 12-15 hours without a preliminary procedure for extracting lipids from algae. Treatment of raw or frozen algae with an aqueous solution of hydrochloric acid upon heating allows the mild acid hydrolysis of lipids to form free fatty acids and the simultaneous recovery of water-soluble polysaccharides.

Hydrolysis of lipids requires compliance with temperature and time conditions. When using room temperatures (18-25 ° C), the process of lipid hydrolysis proceeds slowly (more than 2 days) and not fully (example 4). The resulting mixture contains, along with free fatty acids, triacylglycerols and polar lipids that have not undergone hydrolysis. Heating algae in an acid solution accelerates the process of lipid hydrolysis. It was experimentally obtained that the use of temperatures in the range of 45-50 ° C is sufficient for complete hydrolysis of both reserve and polar lipids of algae, and the optimal hydrolysis time for heating is 4-5 hours, then the algae in an acid solution is left for 12-15 hours without heating (examples 1, 5, 6, 7). Hydrolysis at higher temperatures accelerates the process of lipid hydrolysis, but can lead to deep destruction of water-soluble polysaccharides of algae.

Typically, the hydrolysis of polar and storage lipids isolated from plant and animal raw materials is carried out at 50-70 ° C, and in some cases at higher temperatures, using acid solutions in the range of 0.5-2.5 N. In the hydrolysis of lipids in algae, we took into account the presence of our own water in raw or frozen raw materials, therefore, the amount of acid for hydrolysis was taken higher than that used for the isolation of water-soluble polysaccharides from dry nonfat raw materials (usually 0.05 N). It has been experimentally established that the acid content is in the range of 0.6-0.8 n. it is sufficient for complete hydrolysis of both neutral (triacylglycerols) and polar lipids and does not lead to the destruction of water-soluble polysaccharides (examples 1, 5, 6, 7). During lipid hydrolysis, cell membranes are simultaneously destroyed, which simplifies and facilitates the subsequent extraction with ethyl alcohol of the target products - free fatty acids, sterols, polyphenolic compounds in complex mixtures. Fat-free algae after removing the remaining alcohol from it by air drying is used for the subsequent production of alginic acid salts.

The essence of the method is as follows: kelp (freshly picked or frozen) is poured 0.6-0.8 N. an aqueous solution of hydrochloric acid (with the formation of a “mirror”) and kept at a temperature of 45-50 ° C for 4-5 hours depending on the type of algae and then the algae in the acid solution is left for 12-15 hours without heating. After removal of the aqueous acid solution (extract I), the remainder of the algae is homogenized in 96% ethanol to recover hydrolysis products. The extraction procedure is repeated three times. Alcohol solutions are separated from the remainder of the algae, combined and concentrated to a minimum volume (approximately 1/10 of its initial volume) to obtain extract II, an equal volume of hexane solvent (hexane or nefras) is added to extract II, the mixture is stirred and left to separate into two phases - hexane and water-alcohol.

The hexane phase containing free fatty acids as the main component is concentrated to dryness on a rotary evaporator. The dry residue is dissolved in hexane and the final purification of free fatty acids from impurities is carried out using ultrafast column chromatography under pressure. Silica gel with a particle size of 40 / 100-100 / 160 microns is used as the sorbent. Silica gel in the column is loaded in hexane, after the sorbent has settled, a hexane solution with the desired products is applied to its surface. For sequential elution of free fatty acids, sterols, pigments from a chromatographic column, hexane and a gradient of diethyl ether in hexane are used. Elution of products is carried out with 5-7 column volumes of solvents, collecting fractions of one column volume. Control of elution of the components is performed by thin layer chromatography in the presence of commercial standards of fatty acids and sterols.

The water-alcohol phase is neutralized to pH 7 and filtered. The solution was concentrated to a small volume, dichloromethane (or chloroform) was added, and stirred. After the solution is divided into two phases, the lower dichloromethane fraction is separated. The upper phase is washed again with dichloromethane, after which it is discarded. The resulting dichloromethane extracts are combined, washed with water, filtered and concentrated to obtain a fraction of polyphenolic compounds (PPS), which is further purified by silica gel column chromatography.

An aqueous solution of hydrochloric acid (extract I) is used to extract water-soluble polysaccharides by precipitation with ethyl alcohol. After separating the precipitate of water-soluble polysaccharides by centrifugation or settling in the cold, the water-alcohol supernatant is concentrated to a small volume and again poured into ethanol, exposed to cold to form a second precipitate, after which the supernatant is drained and discarded. Precipitation is combined, purified by dialysis against distilled water from salt impurities.

The remainder of the algae obtained after alcohol extraction is dried in air to remove residual alcohol and used to isolate salts of alginic acids by a known method (RU 2240816 C1, 11.27.2004).

The invention is illustrated by the following examples of specific performance.

Example 1

Extraction 1. 150 g of raw algae (25 g dry) ribbed kelp (Costaria costata [Turn.] Saund.) Is poured 250-300 ml of 0.8 N. an aqueous solution of hydrochloric acid (with the formation of a “mirror”) and kept at a temperature of 50 ° C for 4 hours, after the cessation of heating, the algae is left for 15 hours in an acid solution. When using frozen algae, its mass decreases due to freezing of water to 50 g (25 g of dry algae), therefore, 0.6 n. hydrochloric acid solution.

The aqueous solution of light green acid is drained, the algae is additionally washed with 250-300 ml of drinking water. Aqueous acid solutions are combined (extract I is obtained).

Extraction 2. The remaining tissue is extracted with 400-500 ml of 96% ethanol in a homogenizer for 3-5 minutes. The ethanol extract is drained, the procedure is repeated twice more with a fresh portion of alcohol. Ethanol extracts are combined, filtered and concentrated in vacuo to 80-100 ml (extract 2).

Extraction 3. The remaining fat-free algae is dried to dryness in air and poured into 300 ml of a 1% aqueous solution of sodium bicarbonate, heated for 3 hours at a temperature of 65 ° C to obtain an aqueous solution of sodium salt of alginic acid. After draining solution 1, the remainder of the algae is re-extracted with the same solution to obtain solution 2.

Isolation of target products from extracts:

Isolation of water-soluble polysaccharides.

To isolate a mixture of water-soluble polysaccharides (fucoidan and trace amounts of laminaran), extract I, which is an aqueous solution of hydrochloric acid, is used. The solution (600 ml) is poured into 2.5 l of 96% ethanol. A white flaky suspension is formed. After settling it in the cold, the alcohol solution is drained, the precipitate of water-soluble polysaccharides is washed twice with alcohol and the alcohol solutions are sent for regeneration. The yield of a mixture of water-soluble polysaccharides after drying is about 0.78 g.

Obtaining free fatty acids, sterols, derivatives of chlorophylls and carotenoids.

An equal volume of hexane is added to 80 ml of extract 2, placed in a container with a bottom drain. The mixture is stirred, left for 10 -15 min to separate into two phases - hexane (upper) and water-alcohol (lower).

The hexane phase is separated from the water-alcohol phase and washed additionally with two volumes of water. The water part is discarded. The hexane phase is evaporated on a rotary evaporator under reduced pressure. The yield of crude fatty acids is 0.32 g (a mixture of fatty acids with derivatives of chlorophylls, carotenoids, sterols, fatty acid esters, unsaturated hydrocarbons).

0.32 g of the mixture is dissolved in 50-100 ml of hexane and placed on a chromatographic column with silica gel with a particle size of 40/100 microns (sorbent column height 15-20 cm, column diameter 2 cm). The flow rate of the eluent (within 30-40 ml / min) through the column provide reduced pressure. Hexane and a gradient of diethyl ether in hexane are used sequentially to elute the desired products. Elution of products is carried out with 5-7 column volumes of solvents, collecting fractions of one column volume. Control of the elution of the target products is performed by thin-layer chromatography in the presence of commercial standards of fatty acids, sterols, triacylglycerols. Fractions containing identical substances are concentrated on a rotary evaporator. 0.19 g of free fatty acids is obtained. The yield is about 88% of the amount of fatty acids. The yield of sterol is 0.053 g (90% of the amount of sterols). Derivatives of chlorophylls and carotenoids remain on the column. Elution with a mixture of hexane and diethyl ether gave 0.17 g of product.

Obtaining polyphenolic compounds.

After separation from the hexane phase, the aqueous-alcohol phase is neutralized to pH 7, filtered, concentrated to a volume of about 50 ml, two volumes of dichloromethane (or chloroform) are added, the mixture is stirred. After the solution is divided into two phases, the lower dichloromethane fraction is separated. The upper phase is re-washed with dichloromethane to completely remove the SFC, after which the upper phase is discarded. The dichloromethane fractions are combined, filtered and concentrated. The product yield, which is the fraction containing polyphenolic compounds (PPS), is of the order of 122 mg. The final purification of the PFC fraction is carried out on a column of silica gel with a particle size of 40/100 microns (column height 10-12 cm, column diameter 1 cm). To elute the desired products, chloroform and a gradient of ethanol in chloroform are used. The flow rate of the eluent through the column is provided by reduced pressure. Control of elution of the target products is performed using thin layer chromatography in the presence of commercial standards. The eluates containing the desired products are concentrated on a rotary evaporator to obtain 0.1 g of PPS.

Obtaining sodium salts of alginic acids.

300 ml of an aqueous solution of 1 obtained by extraction of the remainder of the algae with sodium hydrogen carbonate, as described above, is neutralized to pH 7 and poured into 1 liter of ethyl alcohol. Immediately formed white flakes of salts of alginic acid, which are filtered off through a fine-meshed nylon cloth, two more times dissolved in 96% ethanol, freed from alcohol by filtration through a fine-meshed cloth, and dried to a constant weight. The yield of alginic acid salts from solution 1 is 2.46 g.

A similar procedure is carried out with solution 2. The yield of alginic acid salts from solution 2 is 1.39 g.

Example 2. 150 g of ribbed kelp is subjected to the procedures described in example 1, however, the processing of algae is carried out at a temperature of 50 ° C for 2 hours, 0.2 N. aqueous hydrochloric acid. Obtaining salts of alginic acids is carried out as in example 1.

Carrying out the process under these conditions leads only to partial hydrolysis of algal lipids. Free fatty acids make up about 20% of the total lipid. When extracting algae according to the scheme described in example 1, carrying out the extraction of lipids with alcohol, a mixture is obtained containing, in addition to free fatty acids, triacylglycerols, glyceroglycolipids and phospholipids.

The yield of products is: water-soluble polysaccharides - 0.79 g, fatty acids - 0.03 g, a mixture of lipids and pigments - 0.78 g, PPS - 0.1 g, salts of alginic acids - 4.2 g.

Example 3. 150 g of raw ribbed kelp is subjected to the procedures described in example 2, however, the processing of algae is carried out 0.2 N. an aqueous solution of hydrochloric acid at a temperature of 50 ° C for 5 hours and then the algae is left in the acid solution until morning without heating. In this case, lipid hydrolysis takes place by about 50%, which leads to an increase in the yield of free fatty acids.

The yield of products is: water-soluble polysaccharides - about 0.7 g, fatty acids 0.07 g, a mixture of lipids and pigments - 0.68 g, PPS - 0.1 g.

Example 4. 50 g of frozen ribbed kelp is subjected to the procedures described in example 1, however, the processing of algae is carried out at room temperature for 24 hours, 0.6 N. aqueous hydrochloric acid. Obtaining sodium alginate is carried out as in example 1.

As a result of incomplete lipid hydrolysis, only 0.07 g of free fatty acids and 0.36 g of lipids (triacylglycerols, glyceroglycolipids, phospholipids) that have not undergone hydrolysis, and pigments, 0.06 g of PPS are isolated.

Example 5. 50 g of frozen ribbed kelp is subjected to the procedures described in example 1. The treatment of algae is carried out at 45 ° C for 4 hours, 0.6 N. aqueous hydrochloric acid and then left without heating until morning. Obtaining salts of alginic acids is carried out as in example 1.

The yield of products is: water-soluble polysaccharides - about 1.1 g, fatty acids 0.25 g, PPS - 0.2 g, sterols - 0.072 g, salts of alginic acids - 6.5 g.

Example 6. 120 g of raw chicory laminaria (Laminaria cichorioides, Miyabe, family Laminariacea) is subjected to the procedures described in example 1. Processing of algae is carried out at 45 ° C for 4 hours, 0.8 N. aqueous hydrochloric acid. Obtaining salts of alginic acid is carried out as in example 1.

Obtain 0.14 g of free fatty acids, 0.027 g of sterols, 0.1 g of PPS, 2.3 g of water-soluble polysaccharides (a mixture of laminaran and fucoidan), 7.2 g of sodium alginate.

Example 7. 50 g of frozen chicory laminaria is subjected to the procedures described in example 7, however, algae is treated with 0.6 n. an aqueous solution of hydrochloric acid at 45 ° C for 4 hours and then left until morning in an acid solution without heating.

The yield of products is: 0.15 g of free fatty acids, 0.027 g of sterols, 0.095 g of PPS, 2.3 g of water-soluble polysaccharides, 7.4 g of salts of alginic acids.

Claims (2)

1. The method of processing brown algae, which consists in the fact that the raw material is poured into 0.6-0.8 N. an aqueous solution of hydrochloric acid, kept at a temperature of 45-50 ° C for 4-5 hours, leave the algae in an acid solution for 12-15 hours without heating, then the aqueous acid solution is separated to obtain extract I, from which water-soluble polysaccharides are extracted (PS ) by precipitation with 96% ethyl alcohol, then the algae residue is homogenized three times in 96% ethanol, the alcohol extracts are separated from the algae residue, combined and concentrated to a minimum volume to obtain extract II, then an equal volume is added to extract II hexane solvent, the mixture is stirred and left to separate into two phases - hexane and water-alcohol, then the hexane phase, separated from the water-alcohol phase, is dried to dryness, redissolved in hexane and applied to a chromatographic column with silica gel to isolate fatty acids (LC ), sterols and pigment derivatives by sequential elution with hexane and a gradient of diethyl ether in hexane; then the aqueous-alcoholic phase is neutralized and filtered, the solution is concentrated to a small volume, dichloromethane or chloroform is added to extract the fraction of polyphenolic compounds (PPS), then the PPS fraction is washed with distilled water, dried, dissolved in chloroform and PPS is further purified by silica gel column chromatography. 2. The method according to claim 1, in which the remainder of the algae is dried and used to obtain salts of alginic acids.