RU2337571C2 - Method of complex processing sea ware (versions) - Google Patents

Abstract

FIELD: medicine; pharmacology.

SUBSTANCE: method of complex processing of sea ware (Fucus vesiculosus) provides obtaining of a lipide-pigmental complex - a dense extract, cleared mannite, polysaccharide complex - a dry extract of sea ware and sodium alginate. Thus for obtaining of a lipide-pigmental complex, the dry crushed seaweed is extracted with an admixture of methylene chloride with alcohol ethyl in the ratio 94.2:5.86 % strength., concentrated and dried. For obtaining of mannite the algal extraction cake is extracted using the method of a percolation of 85-90 % ethanol with compulsory circulation of an extracting agent in each hour of digestion within 3-5 hours, filtered, concentrated, crystallised, dried and cleared. For obtaining of a dry extract of sea ware the remained extraction cake is extracted using 5-15 % ethanol solution at pH 1-2 within 6-9 hours at 40-60°C, concentrated and dried. Then the extraction cake is extracted using 1.5 % solution of sodium carbonate, treated with sulfuric acid, concentrated, cleared, dried and obtain sodium alginate. As the version, the method of complex processing of seaweed (Fucus serratus) or seaweed (Fucus distichus) provides obtaining of a lipide-pigmental complex using extraction of dry crushed seaweed in an admixture of chloroformium with ethanol in the ratio 88.06:11.9 % strength. Mannite is obtained by extraction of seaweed extraction cakeusing the method of percolation of 85-90 % ethanol at 45-65°C. The dry extract of fucus is prepared by an extraction of the remaining cake using the method of circulating percolation of 10-30 % water ethanol at pH 1-4 within 6-12 hours at 45-75°C. For obtaining of sodium alginate the remaining cake is extracted using the method of percolation of 1.5-4 % solution carbonate at 45-65°C, cleared and dried. As the version, the method of complex processing of knotty wrack (Ascophyllum nodosum) provides obtaining of a lipide-pigmental complex by extraction of seaweed using an admixture of chloroformium with ethanol in the ratio 1:1 with the method of percolation with compulsory circulation of the extraction agent in each hour of digestion within 20-24 hours. Mannite is obtained using extraction of the remaining cake with the method of percolation of 85-95 % ethanol at 65-75°C. A dry extract of rockweep is prepared by extraction of the remaining cake with the method of circulating percolation of 5-15 % water ethanol at pH 4-6, within 12-18 hours at 70-80°C. Then the cake is extracted using the method of percolation of 1.5-2.5 % solution of sodium carbonate at 65-75°C, the extract is treated with the sulfuric acid, the obtained deposit of alginic acid is dissolved in 1.5 % solution of sodium carbonate, concentrated, cleared, dried, thus obtaining sodium alginate polysaccharide.

EFFECT: invention allows increasing output of biologically active substances in the obtained end-products.

5 cl, 10 dwg, 4 tbl

Description

The invention relates to the medical, chemical, pharmaceutical and food industries and relates to a technology for the integrated processing of fucus algae, mainly algae of the Barents Sea, to produce both individual substances and complex preparations in a single technological cycle.

The method suggests using non-traditional types of raw materials as raw materials for producing biologically active substances - fucus algae of the Barents Sea (Fucus vesiculosus, Fucus serratus, Fucus distichus and Ascophyllum nodosum). As final products, one obtains: a lipid-pigment complex in the form of a thick extract, mannitol, a polysaccharide complex in the form of a dry extract, and sodium alginate.

Fucus algae are natural sources of biologically active substances (BAS). They contain mannitol, salts (sodium, calcium, potassium, etc.) of alginic acid, fucoidan, laminaran, lipids, pigments, sterols and others. The qualitative and quantitative characteristics of these biologically active substances of algae vary depending on many factors, such as the type of algae (species-specificity), the stage of development of plants, the conditions for their growth, etc. Therefore, to obtain a biologically active substance with standard characteristics, the above factors must be taken into account.

Fucus vesiculosus (Fucus vesiculosus) is the most studied and widely used species of Fucus algae, due to its ubiquitous distribution, as well as easy accessibility for collection and harvesting. Other types of Fucus algae (Fucus serratus, Fucus distichus and Ascophyllum nodosum) grow on the coast of the Barents Sea, which are not inferior to Fucus vesiculata in the content of biologically active substances, their reserves are sufficient for industrial use.

Lipid-pigment complexes isolated from different types of fucus algae differ in the composition and content of higher fatty acids, pigments, and sterols. The total number of pigments in fucoids growing on the coast of the Barents Sea is highest for Fucus vesiculosus and Fucus serratus, and the lowest for Ascophyllum nodosum, while the content of β-carotene is higher for the algae Fucus serratus and Fucus distichus compared to other species. The highest contents of fucoxanthin and chlorophyll (a + c) were noted for Fucus vesiculosus and Fucus serratus. The total lipid content increases in the series:

A. nodosum> Fucus serratus> F. vesiculosus> F. distichus.

The main fatty acids (FAs) of brown algae are myristic, palmitic, oleic, C ₁₈ and C ₂₀ polyene fatty acids, the ratio of which is different for different species (Khotimchenko S.V. Lipids of macrophytes and herbs, Vladivostok, Dalnauka, 2003) . Ascophyllum nodosum, in comparison with other fucus acids, contains 12.4-12.6% (of the total FA) of myristic acid, while its content in other species does not exceed 10%. The content of palmitic acid in Fucus serratus reaches 26.4% (of the total FA). The ratio of (n-3) and (n-6) series of polyenoic acids is different in the species under consideration: Ascophyllum nodosum and Fucus distichus contain approximately equal amounts, in Fucus vesiculosus and Fucus serratus the level of (n-6) polyunsaturated fatty acids is higher than ( n-3). Significant amounts (2-4%) of rare acids of irregular structure were found in the algae Ascophyllum nodosum - Δ5,11,14-20: 3 and Δ5,11,14,17-20: 4.

Given the above, it becomes obvious that lipid-pigment complexes isolated from different types of fucus algae will vary both in qualitative composition and in the ratio of the main components, and therefore in the pharmacological effect on the human body.

Fucus algae contain three types of polysaccharides: alginic acid, fucoidan and laminaran.

Alginic acid is a heterogeneous substance consisting of fractions of different solubility and composition of uronic acids, mainly from blocks of mannuronic (M) and guluronic (G), and from the changing chains of these two monomers (Nishide E. et al., Changes in M / G ratios of extracted and residual alginate fractions on boiling with water the dried brown alga. // Hydrobiologia. 1996. V.327. N Jul. P.515-518). Blocks of polymannuronic acid add viscosity to alginate solutions, blocks of guluronic acid are responsible for gel strength and specific binding of divalent metal ions (Stokke BT et al. Distribution of uronate residues in alginate chains in relation to alginate gelling. // Macromolecules. 1991. V.24. N 16. P. 463-4345). In different fractions of alginic acid, the content of mannuronic and guluronic acids is different. To a large extent, the properties of alginic acid and its salts depend on the molecular weight. Significant fluctuations in molecular weight (35-1500 kDa) depend on the type of algae and the method of production. Commercial sodium alginate is predominantly isolated from kelp.

Fucoidans are a group of high molecular weight sulfated polysaccharides (molecular weight 100-800 kDa), constructed from residues of α-L-fucopyranose. These biopolymers contain up to 30% ether-bound sulfuric acid. Fucose residues are connected by β-1,2-; β-1,3- and β-1,4-bond. The structural units of fucoidan are presented in figure 1.

The content and structural characteristics of fucoidans vary significantly depending on the type of algae. Differences in the structure of fucoidans of different origin are reflected in the biological activity of these polysaccharides.

It has now been established that fucoidan isolated from F. vesiculosus algae has the following structure (Lewis G. et a1. Commercial production and applications of algal hydrocolloids. University of Washington. Seattle. 1988):

[→ 3) -α-L-Fuc (2SO ₃ ^- ) - (1 → 4) -α-L-Fuc (3,3diSO ₃ ^- ) - (1 →] _n

Fucoidan from F. vesiculosus is characterized by anticoagulant and antithrombotic activity. Fucus vesicle fucoidan is a commercial preparation and its biological effect and structural characteristics are best studied.

Fucoidan from algae F. serratus (Bilan M.I. et al. Structure of fucoidan from the brown seaweeds Fucus serratus L. // Carbohydr. Res. 2006. 341. P.238-245) has the structure:

[→ 3) -α-L-Fuc (2R ₁ R ₂ ) - (1 → 4) -α-L-Fuc (2SO ₃ ^- ) - (1 →] _n

(~ 50%): R ₁ = SO ₃ ^- , R ₂ = H

(~ 50%): R ₁ = H,

R ₂ = α-L-Fuc- (1 → 4) -α-L-Fuc (2SO ₃ ^- ) - (1 → 3) -α-L-Fuc (2SO ₃ ^- ) - (1 →

Fucoidan isolated from F. serratus has a high immunomodulating effect (Green JR. Et al. Binding of monoclonal antibodies to vegetative tissue and fucose-containing polysaccharides of Fucus serratus L. // New Phytol. 1993. 124. N 3. P. 397-408).

The structure of fucoidan from algae F. distichus (Bilan M.I. et al. A highly regular fraction of a fucoidan from brown seaweed Fucus distichys L. // Carbohydr. Res. 2004. 339. P.511-517) was studied:

[→ 3) -α-L-Fuc (2,4-di-SO ₃ ^- ) - (1 → 4) -α-L-Fuc (2SO ₃ ^- ) - (1 →] _n

The polysaccharide from this species of algae significantly reduces the fertility of the mammalian organism (Serrao EA et al. Successful external fertilization in turbulent environments. // Proc. Natl. Acad. Sci. USA. 1996. May. 28. 93. N 11. P.5286 -5290).

Fucoidan from the algae Ascophyllum nodosum (Chevolot L. et al. A disaccharide repeat unit is the major structure in two species of brown algae. // Carbohydr. Res. 2001. 330. P.529-535) has the following structure:

[→ 3) -α-L-Fuc (2SO ₃ ^- ) - (1 → 4) -α-L-Fuc (2,3-diSO ₃ ^- ) - (1 →] _n

Fucoidan from A. nodosum has antitumor and antiproliferative properties (Riou D. et al. An antithcancer and antiproliferation activities of a fucoidan fraction from Ascophyllum nodosum. // Anticancer Res. 1996. 16. P.1213-1218).

Lamaran is a low molecular weight polysaccharide consisting of D-glucopyranose residues connected in linear chains by β-1-3 bonds (may contain weakly branched sections of β-1-6). Up to 75% of the molecules are attached by a β - 1-3 bond to the remainder of D-mannitol. Molecular Weight 3500-5000 Yes (Mayer AM et al. Biological activity in Macrocystis purifera from Argentina: Sodium alginate, fucoidan and laminaran. // XII-th Intern. Seaweed Symp. 1987. V.151-152. P.497-500 ) The structural formula of laminaran is presented in figure 2.

Lamarana are cryoprotectants and anti-cancer agents. The content of laminarans in Fucus algae is small (1-4%), they are part of their water-soluble extracts.

A known method of complex processing of brown algae to obtain iodine-containing and polysaccharide products (RF patent No. 2233104 from 12/09/02). The problem is solved by grinding the raw materials (Japanese kelp), extracting it with an aqueous-alcoholic solution (65-75% ethanol), separating the aqueous-alcoholic extract, settling, and separating the aqueous and lipid fraction. An iodine-containing mannitol complex is obtained from the aqueous fraction, and an iodine-containing lipid complex is obtained from the lipid fraction.

Meal (I) is extracted with a 0.1 N hydrochloric acid solution for 6–10 h at room temperature, then the reaction mixture is heated to a temperature of 45–55 ° С for 4–6 h. The hydrochloric acid extract is neutralized with potassium or sodium carbonate, concentrated in vacuo, and precipitated with 96% ethanol.

Meal (II) is extracted with a solution of sodium carbonate for 10-15 hours at room temperature, then the reaction mass is kept at a temperature of 80-85 ° C, the filtrate is separated, and calcium algylose or sodium algylose is obtained as a biologically active substance. Calcium algylose is obtained by adding a 10% solution of calcium chloride to the filtrate, sodium algylose - by adding sodium carbonate to alginic acid. Alginic acid is precipitated from the filtrate with a 10% hydrochloric acid solution.

The proposed method for the integrated processing of algae allows you to simultaneously obtain several food additives containing biologically active substances: two iodine-containing products - iodine-containing mineral-mannitol and iodine-containing lipid complexes, sodium algylose and calcium algylose, as well as fucoidan concentrate. The resulting nutritional supplements have a wide range of pharmacological properties.

The disadvantage of the developed technology is the long extraction of raw materials and meal when obtaining fucoidan concentrate by an inefficient method (single maceration) without creating the necessary difference in concentrations in the raw material – extractant system. The resulting product is characterized by a low content of fucoidan (33-40%), contains a significant amount of minerals (31-35%). The yield of this product is also low and amounts to 10-12%. Another disadvantage of this method is to obtain dietary supplements with a low content of biologically active substances of algae: 0.7-1.0% iodine, 28-39% mannitol, 7-8% amino acids, minerals 40-50%. The content of these biologically active substances in raw materials (L. japonica) is slightly lower, therefore, the main goal in creating dietary supplements is to obtain biologically active substances concentrates and increase their bioavailability.

A known method of producing biologically active substances from kelp for medical purposes (RF patent No. 2194525 from 07/16/01). According to the method, the crushed raw materials (sugar kelp) are degreased with chloroform in the ratio of raw materials: extractant 1:20 for 1.5 hours, sequentially extracted with 90-96% ethanol, hot water and 1.5% sodium carbonate solution. After filtration, ethanol extraction is dialysed through a cellophane membrane, concentrated and mannitol is crystallized from it. Water extraction is precipitated three times at a temperature of 4 ° C, dialyzed, evaporated, precipitated from it with 90% ethanol protein-polysaccharide preparation (BOD) "Laminaride-SB", which is dried by freeze drying. After pretreatment with concentrated sulfuric acid, cooling at 4 ° C and dialysis, sodium alginate is precipitated from carbonate extraction, lyophilized. This method has several differences and disadvantages compared with the claimed method:

1) sugar kelp is used as raw material;

2) when degreasing raw materials with chloroform, valuable biologically active substances are removed - lipids, pigments, sterols;

3) for processing raw materials with chloroform, a low-efficiency method of single maceration at room temperature is used;

4) the extraction of mannitol is carried out in the Soxhlet apparatus by the method of circulating extraction, in which the extraction occurs in the upper part of the apparatus at an extractant temperature of not higher than 20 ± 5 ° C. It is known that the yield of mannitol increases with increasing temperature of the ethanol extractant;

5) the extract containing the polysaccharide-protein complex is maintained at a reduced temperature (4 ° C) three times, while the proteins precipitate, which leads to the loss of the target components of the complex;

6) 90% ethanol is used to precipitate the polysaccharide-protein complex and sodium alginate, which leads to significant losses of polysaccharides (more than 30%). In addition, in an extract containing a polysaccharide-protein complex, after precipitation with ethanol, a significant amount of laminaran is soluble in ethanol with a concentration below 60%;

7) one of the target products, a polysaccharide-protein preparation, contains fucoidan with a main chain built of only 1 → 3-linked fucose residues, as it is isolated from kelp algae (Usov AI, et al. Algae polysaccharides 53. Brown algae Laminaria saccharina (L.) Lam. As a source of fucoidan. // Bioorg. Chemistry. 1998. V.24. S.437-445). Fucoidans isolated from fucus algae are based on a polysaccharide chain of alternating 3- and 4-linked fucose residues. Differences in the structure of fucoidans, representatives of laminaria and fucus algae, are interrelated with the pharmacological effects of these polysaccharides.

A known method of complex processing of brown algae to obtain preparations for medicine and cosmetology (RF patent No. 2240816 from 07.28.03). The method consists in the fact that plant materials (brown algae) are treated with ethanol for 20-24 hours at a temperature of 40-60 ° C, the extraction is separated, the ethanol is distilled off, a concentrate of biologically active substances and low molecular weight compounds is obtained, then the meal is dried and extracted. hydrochloric acid solution for 10-14 hours at a temperature of 20-25 ° C with stirring. The extract is concentrated on an ultrafiltration unit, neutralized to pH 6.0 with sodium hydroxide solution and evaporated. A concentrate is obtained - polysaccharide-1, containing a mixture of laminaran and fucoidan, successively precipitated with ethanol fucoidan (F1) and laminaran (L1). Algae meal is sequentially extracted twice with hot water at pH 3.5-4.0 with stirring for 2-4 hours and 1-2 hours, respectively. The extracts are combined, concentrated on an ultrafiltration unit, evaporated. Get polysaccharide-2, which is a mixture of laminaran, fucoidan and polymannuric acid. Then the pH of the extract was adjusted to a value of 2.0-2.5, the precipitate of polymannuric acid was separated by centrifugation. To obtain the polymannuric acid salt, the precipitate is dissolved in a minimal volume of sodium hydroxide or ammonium oxalate, or calcium hydroxide, or magnesium hydroxide, then the solution is neutralized and the polymannuric acid (M) salt is precipitated by adding two volumes of ethanol, the precipitate is washed with ethanol and dried. The supernatant is neutralized, ethanol is subsequently precipitated with fucoidan (F2), laminaran (L2). Algae meal is extracted with a solution of sodium hydroxide (or calcium, or magnesium) or ammonium oxalate at pH 8-9 for 2-4 hours at a temperature of 55-65 ° C, the extraction is concentrated on an ultrafiltration unit, neutralized and ethanol precipitated with polysaccharide-3, which represents alginic acid salt (A), which is washed with ethanol and dried.

This method is designed for the complex processing of the Far Eastern types of brown algae (L. cichorioides, L. japonica, F. evanescence, etc.), however, the authors argue that the method is universal and can be applied to other types of brown algae to produce in each case a specific set of acidic and neutral polysaccharides. It should be noted that when applying this scheme for fucus algae of the Barents Sea, it is inappropriate to obtain individual laminarana, because in these species, their content is low (0.5-3%), it is also doubtful to obtain polymannuronates.

A feature of this method is the use of algae in fresh or frozen form. Processing of fresh algae must be carried out within the next 48 hours, as processes of enzymatic hydrolysis of polysaccharides, oxidation of lipids and pigments begin very quickly. For this purpose, it is necessary to ensure quick delivery of raw materials to the processing plant, which is often impossible. For frozen algae, a strict storage regime must be observed, their defrosting is unacceptable, because at the same time, there will be a loss of the target components - water-soluble substances (fucoidan, laminaran, vitamins, amino acids, etc.).

The main disadvantages of the method are its multi-stage and the use of ineffective extraction methods - single or double maceration (infusion).

Closest to the claimed invention is a method for the integrated processing of fucus algae Fucus vesiculosus (Obluchinskaya ED Complex use of brown algae; Ros. Chem. Zh., 2004, vol. XLVII, No. 3, p.136-142). The method consists in the fact that dry thalli of algae, namely vesicular fucus, are ground and sequentially extracted with an azeotropic mixture, ethanol, a water-alcohol mixture and sodium hydroxide. To obtain a thick extract of bubbly fucus, the crushed algae are extracted with an azeotropic mixture in a Soxhlet apparatus, concentrated in a vacuum on a rotary evaporator, dried in a vacuum on a rotary evaporator, and dried in a vacuum oven. To obtain mannitol, the meal is extracted with 96% ethanol by the Soxhlet circulating extraction method, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried and purified by column chromatography using alumina. Then the remaining meal is extracted by circulating percolation with an aqueous-alcohol mixture of 5-30% aqueous ethanol, concentrated on an ultrafiltration apparatus with simultaneous dialysis, dried and a polysaccharide complex is obtained - a dry extract of Fucus. To obtain sodium alginate, meal is extracted by bismaceration with a 2% sodium carbonate solution, treated with sulfuric acid, dissolved in a 1.5% sodium carbonate solution, purified by dialysis and dried. This technology has been developed and is used to process only bubbly fucus.

The objective of the invention is to develop the above method and improve methods for the integrated processing of other types of fucus algae to obtain biologically active substances with standard characteristics applicable in medicine, the pharmaceutical industry and the food industry.

Technical results are: obtaining a specific set of biologically active substances with standard characteristics from Fucus algae of the species Fucus vesiculosus, Fucus serratus, Fucus distichus and Ascophyllum nodosum; an increase in the yield of target products by 5-17% depending on the types of algae while increasing the degree of their purity, including from fucus vesiculum by 7-17%, namely: LPC - by 15-17%, mannitol - by 7%, fucoidan - by 10-17%, sodium alginate - by 8-16%; from bilateral fucus: LPC - by 5%, mannitol - by 4-7%, fucoidan - by 14-17%, sodium alginate - by 6-15%; from cucumber fucus: LPC - by 9%, mannitol - by 6-8%, fucoidan - by 9-14%, sodium alginate - by 6-15%; and ascophyllum nodosum: LAB by 15-24% with a chlorophyll content of at least 8%, mannitol 11-13 %%, polysaccharide complex 18-24% and sodium alginate 12-15%.

The method, as well as known, is that dry thalli of algae are crushed, extracted with a mixture of organic extractants, concentrated in a vacuum on a rotary evaporator, dried in a vacuum on a rotary evaporator, dried in a vacuum oven to obtain a lipid-pigment complex - thick extract , then sequentially extracted with ethanol, a water-alcohol mixture and sodium hydroxide to obtain mannitol, a dry extract and sodium alginate.

The technical result is achieved by the fact that:

1. For the processing of Fucus algae species of vesicular fucus (Fucus vesiculosus):

to obtain a lipid-pigment complex (thick Fucus extract), extraction is carried out with a mixture of methylene chloride and ethyl alcohol in a ratio of 94.2: 5.86% in a Soxhlet apparatus, to obtain mannitol, algal meal is extracted by percolation with 85-90% ethanol with forced circulation of the extractant through each hour of infusion for 3-5 hours, to obtain a dry extract of Fucus, meal is extracted with 5-15% ethanol solution at pH 1-2 and the infusion time of 6-9 hours at a temperature of 40-60 ° C.

2. For the processing of fucus algae of the species of dentate fucus (Fucus serratus) or bilateral fucus (Fucus distichus):

to obtain a lipid-pigment complex (thick Fucus extract), the extraction is carried out with an azeotropic mixture of methylene chloride with ethyl alcohol in a ratio of 94.2: 5.86% or an azeotropic mixture of chloroform with ethanol in a ratio of 88.06: 11.9% in a Soxhlet apparatus, then algal meal is extracted by percolation with 85-95% ethanol at a temperature of 45-65 ° C, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried, purified by column chromatography using alumina and purified mannitol is obtained, then the remaining meal is extracted by circulating percolation with 10-30% aqueous ethanol at pH 1-4, the infusion time is 6-12 hours at a temperature of 45-75 ° C, it is purified by vacuum concentration followed by dialysis through a cellophane membrane or concentration on ultrafiltration apparatus with simultaneous dialysis, dried by lyophilization or in a vacuum oven and get a polysaccharide complex - a dry extract of Fucus, then the meal is extracted by percolation with a 1.5-4% sodium carbonate solution at a rate a temperature of 45-65 ° C, the extract is treated with sulfuric acid, the obtained alginic acid precipitate is dissolved in a 1.5% sodium carbonate solution, concentrated on a rotary evaporator, purified by dialysis through a cellophane membrane, dried in a vacuum oven or lyophilization, and sodium alginate polysaccharide is obtained .

3. For the processing of Fucus algae of the Ascophyllum nodosum (Ascophyllum nodosum):

to obtain a lipid-pigment complex (thick ascofillum extract), extraction is carried out with a mixture of chloroform and ethanol in a 1: 1 ratio by percolation with forced circulation of the extractant after every hour of infusion for 20-24 hours, then algal meal is extracted by percolation with 85-95% ethanol at a temperature of 65-75 ° C, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried, purified by column chromatography using alumina and purified mannitol is obtained, then the remaining meal is extracted by circulating percolation with 5-15% aqueous ethanol at pH 4-6, the infusion time is 12-18 hours at a temperature of 70-80 ° C, the extract is filtered, concentrated under vacuum on a rotary evaporator, dialyzed in water purified through a cellophane membrane, dried by lyophilization or in a vacuum oven and get a polysaccharide complex - a dry extract of ascofillum, then the meal is extracted by percolation with a 1.5-2.5% solution of sodium carbonate at a temperature of 65-75 ° C, the extract is treated with With sulfuric acid, the resulting alginic acid precipitate is dissolved in a 1.5% sodium carbonate solution, concentrated on a rotary evaporator, purified by dialysis in water purified through a cellophane membrane, dried in a vacuum oven or lyophilization, and sodium polysaccharide is obtained.

Our proposed method involves the use of dried raw materials (drying at a temperature not exceeding 60 ° C).

The essence of the method and the influence of the essential features of the formula on the achieved result is explained for each type of fucus algae:

1. The use of algae Fucus vesiculosus (vesiculus fucus) as a raw material. Obtaining lipid-pigment complex LPK. Thalli of dry algae were crushed, extracted in a Soxhlet apparatus with an azeotropic mixture (methylene chloride - ethyl alcohol in a ratio of 94.2: 5.86%) to the maximum depletion of raw materials. At the end of the extraction process, the extract was concentrated in vacuo on a rotary evaporator and dried in a vacuum oven. The yield of the timber industry amounted to 3.20 ± 0.39% (the yield of timber from the content in raw materials - the technological yield - 88.09 ± 0.89%). The resulting product is a thick extract of brown-green color, oily in appearance, having a peculiar smell. Qualitative analysis by thin layer chromatography (Silufol brand 254) showed the presence of β-carotene (Fluka), α-tocopherol (Fluka), β-sitosterol (Fluka), fucosterol (Fluka). Quantitative analysis of chlorophyll was carried out spectrophotometrically using Getri reagent as a calibration solution (L. Sirenko et al. Methods of physiological and biochemical study of algae in hydrobiological practice, Kiev: Naukova Dumka. 1975). The content of chlorophyll in the forest from bladder fucus was 8.63 ± 0.35%.

Compared with the analogue, the infusion time was increased from 1 to 5 hours, which leads to an increase in the yield of woody complex from 71.17 (in the analogue) to 88.09%, as well as an increase in the chlorophyll content from 5.5-6.2 to 8.63 % The increase in infusion time and the use of a certain ratio of the azeotropic mixture allows to obtain a significant quantitative increase in the technical result.

The results, shown in Fig. 3 and Table 4, show that using the extraction in the Soxhlet apparatus with an azeotropic mixture of methylene chloride - ethyl alcohol in a ratio of 94.2: 5.86%, it was possible to increase the technological yield of the woodworking complex by 15-17%, this is confirmed high value of technological output (88.1 ± 0.9%), as well as qualitative and quantitative characteristics of timber industry.

Getting mannitol. To obtain mannitol, we used algal meal remaining after isolation of the timber complex. Unlike the analogue, to obtain mannitol according to the claimed method, a percolation method with forced circulation of the extractant is used, extraction is carried out with 85-90% ethanol at a temperature of 45-65 ° C for 3-5 hours, for which circulation was carried out every 1 hour for circulation extractant. In the analogue, the circulation extraction method is used in the Soxhlet apparatus at a temperature of 20 ° C, ethanol 95% as an extractant. Figure 4 presents data on the influence of the extraction method on the output of mannitol. It can be seen from them that when using the proposed method, the mannitol yield increased by at least 7% compared to the analogue and amounted to 99.2%, therefore, it was possible to more fully extract the target component - mannitol.

Then the extract was filtered, concentrated in vacuo on a rotary evaporator. Then crystallization was carried out at 0 ÷ + 4 C for 24 hours. The precipitated mannitol crystals were filtered on a putsch filter and dried. Received technical mannitol. Next, mannitol was purified by column chromatography using alumina. The resulting mannitol was a white crystalline powder, the physicochemical characteristics of which are presented in Table 1.

The proposed method for producing mannitol was compared with the traditional method used at the Arkhangelsk experimental industrial algae plant (AOPVK), as well as by the method of circulation extraction. Mannitol is obtained at AOPVK by maceration with heating at the boiling point of an extractant - ethyl alcohol with a concentration of 80-96%. The research results presented in Fig. 4 confirm the high efficiency of using the percolation method compared to existing methods: the mannitol yield increased by 7-10% (compared to circulation extraction) and 30-34% (compared to maceration).

Obtaining the polysaccharide complex "Fucoidan-P" (dry extract). In an analogue, the algae meal was extracted by circulating percolation with 5-30% aqueous ethanol without heating. Technological yield 83.37%. The content of the main active ingredient of fucoidan is 71% (determined by the formula Ф = amount of fucose x2, see table 2).

According to the claimed method, the algae meal was extracted by circulating percolation with 5-15% aqueous ethanol, pH = 1-2. The infusion time is 6-9 hours, the temperature is 40-60 ° C. The range of applied extractant concentrations was reduced, the reaction of the medium was changed from neutral to acidic, the optimal conditions for the extraction — time and temperature conditions — were determined. Thus, the percolation conditions of fucoidan differ from the analogue in temperature, pH, and time. It is this optimal selection of all extraction conditions that provided a quantitative increase in the yield of fucoidan of 10-17%, not known from the prior art, amounting to 93-99%. Evidence of the effect of just such extraction modes when using the known circulating percolation method on the output of fucoidan is presented in Figs. 8 and 9.

Then the extract was filtered, concentrated in vacuo on a rotary evaporator. The concentrated extract was dialyzed in purified water through a cellophane membrane. Dried in a vacuum dryer or lyophilization. The polysaccharide complex "Fucoidan-P" is a powder from light brown to dark brown in color, containing fucoidan and laminaran, readily soluble in water with the formation of viscous brown solutions, the physicochemical characteristics of which are presented in Table 2.

Percolation extraction was carried out according to the following procedure: the cake was moistened with extractant, loaded into the percolator layer by layer, filled with extractant until a “mirror” was formed, pressed with a load and insisted for 6-9 hours. The extraction was carried out with forced circulation of the extractant (10% ethanol), for which purpose after 1 hour, the extractant was circulated, the extract was filtered and the content of fucoidan was determined in it.

The determination results presented in figure 5, clearly demonstrate the advantage of the percolation method. The fucoidan content in the extracts obtained by this method is 3–6% higher (20–40% in terms of the content in the feed). A significant effect on the output of fucoidan has an increase in temperature. When extracting bubbly fucus by percolation, the optimal range is 40-60 ° C, at which a high yield of fucoidan is achieved - 93-99% (in terms of the content in the feed). A further increase in temperature does not lead to a significant increase in the output of fucoidan.

Obtaining sodium alginate. According to the analogue, the meal was extracted by the bismaceration method with a 2% sodium carbonate solution at a temperature of 50-60 ° C. The extract was treated with sulfuric acid, an alginic acid precipitate formed, which was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven. The technological yield of sodium alginate was 81.73 ± 1.69%, the purity was 92.17%.

Unlike the analogue according to the claimed method, the meal was extracted by the method of circulating percolation with a 1.5-2.5% solution of sodium carbonate at a temperature of 40-60 ° C. The extract was treated with sulfuric acid, an alginic acid precipitate formed, which was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven or lyophilized. The yield of sodium alginate was 18-19.8% (Fig.6), which in terms of the alginate content in the feed was 89-97%, which exceeds this indicator in the analogue by 8-16%, the purity of the target component is also higher compared with the analogue and amounted to 96.1% (table 3). The Box-Wilson method of mathematical planning determined the optimal range of extractant concentration and temperature, which was used to obtain sodium alginate from bubbly fucus according to the claimed method: a 1.5-2.5% sodium carbonate solution at a temperature of 40-60 ° C by the circulation method percolation.

Thus, the replacement of the extraction method and determination of the optimum concentration of the extractant and temperature allowed to increase the yield of the target product and its purity in the complex processing of raw materials, which is not known from the prior art. Figure 6 presents the results of the analysis of the content of alginic acid in extracts obtained by various methods from meal. The obtained results confirm the advantage of the percolation method compared to the traditional method - maceration for extraction of alginic acid. Therefore, to obtain sodium alginate from a meal of fucus vesiculus, the percolation method was chosen.

2. The use of Fucus distichus algae (bilateral fucus) as a raw material. Obtaining lipid-pigment complex LPK. Thalli of dry algae were crushed, extracted in a Soxhlet apparatus with an azeotropic mixture (chloroform - ethanol in a ratio of 88.06: 11.9%) until the raw materials were depleted. At the end of the extraction process, the extract was concentrated in vacuo on a rotary evaporator and dried in a vacuum oven. The output of the timber industry amounted to 3.1 ± 0.6% (technological yield 76.3%). The resulting product is a thick extract of brown-green color, oily in appearance, having a peculiar smell. Qualitative analysis by thin layer chromatography (Silufol brand 254) showed the presence of β-carotene (Fluka), α-tocopherol (Fluka), β-sitosterol (Fluka), fucosterol (Fluka). Quantitative analysis of chlorophyll was carried out spectrophotometrically using Getri reagent as a calibration solution (Sirenko et al., 1975). The content of chlorophyll in the LPC from bilateral fucus was 5.8 ± 0.5%.

To compare the obtained data with existing methods, the LPC was obtained by maceration with heating. The results shown in figure 3 show that the use of the method of circulating extraction led to an increase in the technological yield of the timber complex by 5%.

Getting mannitol. To obtain mannitol, we used algal meal remaining after isolation of the timber complex. Meal was extracted by percolation with ethanol 85-95% at a temperature of 45-65 ° C. Then the extract was filtered, concentrated in vacuo on a rotary evaporator. Then, crystallization was carried out at 0–4 ° C for 24 hours. The precipitated mannitol crystals were filtered on a suction filter and dried. Received technical mannitol. Next, mannitol was purified by column chromatography using alumina. The resulting mannitol was a white crystalline powder, the physicochemical characteristics of which are presented in Table 1.

In the analogue, the circulation extraction method is used in the Soxhlet apparatus at a temperature of 20 ° C, ethanol 95% as an extractant. The research results presented in Fig. 4 confirm the high efficiency of the percolation method in comparison with existing methods: the mannitol yield increased by 4-7% (compared with circulation extraction) and by 27-31% (compared with maceration).

Obtaining the polysaccharide complex "Fucoidan-D" (dry extract). Algae meal was extracted by circulating percolation with 10-20% aqueous ethanol, pH = 1-2. The infusion time is 6-9 hours, the temperature is 50-70 ° C. Then the extract was filtered, concentrated in vacuo on a rotary evaporator. The concentrated extract was dialyzed in purified water through a cellophane membrane. Dried in a vacuum dryer or lyophilization. The polysaccharide complex "Fucoidan-D" is a powder from light brown to dark brown in color, containing fucoidan and laminaran, readily soluble in water with the formation of brown viscous solutions, the physicochemical characteristics of which are presented in Table 2.

When searching for optimal conditions, a series of experiments was carried out with various ranges of the above factors, including those used in the analogue. Some experimental results are shown in FIG. 10. From the presented data it follows that the combined influence of factors led to an increase in the technological yield of fucoidan to 98%, which exceeded the corresponding figure in the analogue by 14-17%.

Obtaining sodium alginate. Meal was extracted by percolation with a 1.5-2.5% sodium carbonate solution at a temperature of 45-55 ° C. The extract was treated with sulfuric acid, an alginic acid precipitate formed, which was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven or lyophilized. The obtained target product, sodium alginate, was a powder or thin plates of a light cream color, odorless. Physico-chemical characteristics of sodium alginate are presented in table.3.

The yield of sodium alginate was 23-27% (see table 4), which in terms of the content in raw materials was 88-97%, which exceeds this indicator in the analogue by 6-15%. Thus, the achievement of the technical result is ensured precisely by the totality of all the essential features indicated in 1 independent and 5 dependent claims.

3. Use as a raw material of algae Fucus serratus (Fucus serratus). Obtaining lipid-pigment complex LPK. Thalli of dry algae were crushed, extracted in a Soxhlet apparatus with an azeotropic mixture (methylene chloride - ethanol in a ratio of 94.2: 5.86%) to the maximum depletion of raw materials. At the end of the extraction process, the extract was concentrated in vacuo on a rotary evaporator and dried in a vacuum oven. The yield of the timber industry amounted to 2.05 ± 1.00% (technological yield 80.5%). The resulting product is a thick extract of brown-green color, oily in appearance, having a peculiar smell. Qualitative analysis by thin layer chromatography (Silufol brand 254) showed the presence of β-carotene (Fluka), α-tocopherol (Fluka), β-sitosterol (Fluka), fucosterol (Fluka). Quantitative analysis of chlorophyll was carried out spectrophotometrically using Getri reagent as a calibration solution (Sirenko et al., 1975). The content of chlorophyll in the FPC from dentate was 8.53 ± 1.1%.

To compare the obtained data with existing methods, the LPC was obtained by maceration with heating (item 1). The results, reflected in figure 3, show that the use of the method of circulating extraction led to an increase in the technological yield of the timber complex by 16-19%.

Getting mannitol. To obtain mannitol, we used algal meal remaining after isolation of the timber complex. Meal was extracted by percolation with ethanol 85-95% at a temperature of 45-65 ° C. Then the extract was filtered, concentrated in vacuo on a rotary evaporator. Then, crystallization was carried out at 0–4 ° C for 24 hours. The precipitated mannitol crystals were filtered on a suction filter and dried. Received technical mannitol. Next, mannitol was purified by column chromatography using alumina. The resulting mannitol was a white crystalline powder, the physicochemical characteristics of which are presented in Table 1.

In contrast to the analogue for producing mannitol according to the claimed method, the method of percolation with forced circulation of the extractant is used, the extraction is carried out with 85-95% ethanol at a temperature of 45-65 ° C. In the analogue, the circulation extraction method is used in the Soxhlet apparatus at a temperature of 20 ° C, ethanol 95% as an extractant. Figure 4 presents data on the influence of the extraction method on the output of mannitol from the fucus gear. It can be seen from them that when using the proposed method, the mannitol yield increased by 6-8% compared with the analogue, therefore, it was possible to more fully extract the target component - mannitol. Thus, it was the choice of the characteristics given in the formula that made it possible to obtain the indicated technical result.

Obtaining the polysaccharide complex "Fucoidan-S" (dry extract). Algae meal was extracted by circulating percolation with 20-30% aqueous ethanol, pH = 2-4. The infusion time is 9-12 hours, the temperature is 65-75 ° C. Then the extract was filtered, concentrated in vacuo on a rotary evaporator. The concentrated extract was dialyzed in purified water through a cellophane membrane. Dried in a vacuum dryer or lyophilization. The polysaccharide complex "Fucoidan-S" is a powder from light brown to dark brown, containing fucoidan and laminaran, readily soluble in water with the formation of brown viscous solutions, the physicochemical characteristics of which are presented in Table 2.

The percolation conditions of fucoidan differ from the analogue in temperature, pH, and brewing time. The greatest technological yield from denticulate fucus while improving the quality characteristics of the dry fucus extract was obtained precisely under the above extraction conditions specified in dependent paragraph 6 of the formula, which is confirmed by examples 5 and 6 shown in table 4. From the data presented in Fig.9, it follows that the combined influence of factors led to an increase in the technological yield of fucoidan to 94-99%, which also exceeded the corresponding figure in the analogue by 9-14% and is unknown from the prior art.

Obtaining sodium alginate. Meal was extracted by percolation or bismaceration with a 2-4% solution of sodium carbonate at a temperature of 55-65 ° C. The extract was treated with sulfuric acid, an alginic acid precipitate formed, which was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven or lyophilized. The obtained target product, sodium alginate, was a powder or thin plates of a light cream color, odorless. Physico-chemical characteristics of sodium alginate are presented in table.3. From the data presented in Tables 3 and 4 (pp. 7-8, 11), it is obvious that a high yield of alginate is obtained - 18.9-22.27% (which, in terms of the content in raw materials, was 91-95%) and the qualitative characteristics of the obtained target products (purity of sodium alginate up to 97%) that meet the requirements of pharmaceutical standards.

4. The use of Ascophyllum nodosum algae (Ascophyllum nodosum) as a raw material. Obtaining lipid-pigment complex LPK. Dry algae thalli were crushed, extracted by percolation with forced circulation of the extractant after each 1 hour of infusion at a temperature of 20 ± 2 ° C with a mixture of chloroform - ethanol at a ratio of 1: 1, the infusion time was 20-24 hours. After the extraction process was completed, the extract was concentrated in vacuo at rotary evaporator and dried in a vacuum oven. The yield of the timber industry was 1.15 ± 0.66% (technological yield 88.4%). The resulting product is a thick extract of black-green color, oily in appearance, having a peculiar smell. Qualitative analysis by thin layer chromatography (Silufol brand 254) showed the presence of β-carotene (Fluka), α-tocopherol (Fluka). Quantitative analysis of chlorophyll was carried out spectrophotometrically using Getri reagent as a calibration solution (Sirenko et al., 1975). The content of chlorophyll in the timber from Fucus dentus was 10.25 ± 2.23%.

A study of the influence of the extraction method and the extractant on the yield of LPC from ascophyllum nodosum showed that the use of the method of circulating extraction with various extractants did not lead to a significant increase in the yield of LPC (see Fig. 7). Therefore, to obtain this target component, the method of percolation with a mixture of chloroform and ethanol in a ratio of 1: 1 with forced circulation of the extractant after every hour of infusion for 20-24 hours was applied. It is this selection of quantitative ratios of known components of extractants and infusion time and the application of this extraction method - percolation, that leads to a quantitative increase in the yield of woody complex by 15-24% and a high technical result.

Getting mannitol. To obtain mannitol, we used algal meal remaining after isolation of the timber complex. Meal was extracted by percolation with ethanol 85-95% at a temperature of 65-75 ° C.

Then the extract was filtered, concentrated in vacuo on a rotary evaporator. Then, crystallization was carried out at 0–4 ° C for 24 hours. The precipitated mannitol crystals were filtered on a suction filter and dried. Received technical mannitol. Next, mannitol was purified by column chromatography using alumina. The resulting mannitol was a white crystalline powder, the physicochemical characteristics of which are presented in Table 1.

Figure 4 presents data confirming the influence of the selected extraction methods on increasing the yield of mannitol to 11-13% compared with the method used in the analogue. Example 7 of Table 4 shows the modes of percolation of algae meal with ethanol 85% at a temperature of 75 ° C with a brewing time of 4 hours. The mannitol yield was 5.05%, which is 13% higher than when using the circulating extraction method in the Soxhlet apparatus with 95% ethanol. With an increase in ethanol concentration to 95% with a simultaneous decrease in temperature to 65 ° C (see example 8), the mannitol yield slightly decreased and amounted to 4.41%, but still it was 11% higher than that of the analogue. Thus, the use of a more efficient extraction method for this type of algae using the temperature and ethanol concentration ranges specified in the formula led to the achievement of the above technical result — an increase in mannitol yield by 11–13%.

Obtaining the polysaccharide complex "Fucoidan-A" (dry extract). Algae meal was extracted by circulating percolation with 5-15% aqueous ethanol, pH = 4-6. The infusion time is 12-18 hours, the temperature is 70-80 ° C. Then the extract was filtered, concentrated in vacuo on a rotary evaporator. The concentrated extract was dialyzed in purified water through a cellophane membrane. Dried in a vacuum dryer or lyophilization. The polysaccharide complex "Fucoidan-A" is a powder from light brown to dark brown in color, containing fucoidan, readily soluble in water with the formation of viscous brown solutions, the physicochemical characteristics of which are presented in Table 2.

It differs from the analogue in the following:

- the duration of infusion is increased from 6-12 to 12-18 hours, since as a result of the studies, an increase in the time of infusion has a significant effect on the output of fucoidan from ascophyllum nodosum;

- the temperature of infusion is increased from 40-60 ° C, used in the analogue, to 70-80 ° C, since it was at these temperatures that it was possible to achieve the completeness of extraction during the processing of this type of algae;

- the concentration of aqueous ethanol is reduced to 5-15%. The use of a higher concentration of aqueous ethanol - from 5 to 30%, as in the analogue, did not allow to obtain even those results that are indicated in the analogue and actually obtained during the processing of bubbly fucus;

- concentration was carried out under vacuum on a rotary evaporator, and in an analog, on an ultrafiltration apparatus with simultaneous dialysis;

- drying is carried out either lyophilically, as in the analogue, or in a vacuum drying oven. The extracts obtained according to these schemes contained a high fucoidan value and lower ash content, which improves the qualitative and quantitative characteristics of the dry extract.

Table 4 shows examples confirming that the selected methods and modes of extraction increase the yield of the polysaccharide complex to 14%. From the data presented in Fig.9, it follows that the combined influence of the above factors led to an increase in the technological yield of fucoidan to 94-95%, which exceeded the corresponding figure in the analogue by 18-24% and is not known from the prior art.

Obtaining sodium alginate. Meal was extracted by percolation with a 1.5-2.5% solution of sodium carbonate at a temperature of 65-75 ° C. The extract was treated with sulfuric acid, an alginic acid precipitate formed, which was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven or lyophilized. The obtained target product, sodium alginate, was a powder or thin plates of cream color, odorless. Physico-chemical characteristics of sodium alginate are presented in table.3.

It differs from its analogue in the following:

- meal was extracted by percolation with a 1.5-2.5% solution of sodium carbonate at a temperature of 65-75 ° C. Traditionally, alginates from bubbly fucus or from kelp are obtained by maceration of raw materials at a temperature of 50 ° C. In an analogue, the bismateration method was used with a 2% sodium carbonate solution at a temperature of 50-60 ° C. A comparative study showed that the optimal values that provide an increase in the yield of alginate from ascorbid nodosum to 13-14% are in the temperature range 65-75 ° C. As follows from the examples in table 4, the increase in the yield of sodium alginate is higher with increasing concentration of the solution and increasing temperature. This made it possible to increase the purity of sodium alginate and to obtain an increase in the yield of sodium alginate by 12-15%. Thus, such a selection of quantitative ratios of known extractant components, temperature conditions and infusion time leads to a quantitative increase in the technical result: obtaining a specific set of biologically active substances with standard characteristics from a new type of raw material - Ascophyllum nodosum (Ascophyllum nodosum) with an increase in yield: LPC by 15-24 % with a chlorophyll content of at least 8%, mannitol up to 9%, polysaccharide complex 17-24% and sodium alginate 12-15%.

The invention is characterized by examples (see table 4).

Table 1 The results of the analysis of physico-chemical parameters of mannitol Indicator Mannitol - the standard sample Mannitol obtained by complex processing of raw materials F. vesiculosus F. distichus F. serratus A. nodosum Mass fraction 95.7 99.0 99.5 97.9 95.5 the main substance. % 166 166 165 165.5 166 Melting point, ° С Specific rotation [α] _D ^{20 of} 10% mannitol solution in sodium tetraborate solution +23 +23.5 +23.7 +24.1 +23.5 Ash content,% 0.05 0.05 0,03 0.05 0.05 Loss in mass upon drying,% 0.1 0.1 0,07 0.09 0.1 Mass fraction of heavy metals,% no more than 0,001 no more than 0,001 no more than 0,001 no more than 0,001 no more than 0,001

table 2 The results of the analysis of physico-chemical parameters of the polysaccharide complex Indicators Fucoidan-P Fucoidan-D Fucoidan-S Fucoidan-A Description Light brown or cream powder Solubility Soluble in water, insoluble in organic solvents Authenticity: polysaccharides + + + + fucoidan + + + + laminaran + + + + Mass loss on drying 4.79 ± 0.55% 3.81 ± 1.12% 4.60 ± 1.50% 2.94 ± 2.00% Sulphated Ash 22.76 ± 1.39% 21.04 ± 1.19% 20.37 ± 1.65% 27.50 ± 2.22% and heavy metals no more than 0,001% no more than 0,001% no more than 0,001% no more than 0,001% Mass fraction of basic substances: Fucoidan 85.06 ± 0.80% 71.45 ± 2.24% 69.09 ± 1.95% 65.15 ± 3.00% Lamaran 3.70 ± 1.34% 9.77 ± 1.90% 18.75 ± 2.13% - Monosaccharides: Xylose 1.18 ± 0.55% 2.54 ± 0.10% 1.80 ± 0.94% 3.35 ± 1.00% Mannose 0.72 ± 0.22% 0.35 ± 0.05% 0.75 ± 0.10% - Galactose 1.67 ± 0.60% 1.15 ± 0.05% 2.34 ± 0.13% -

Table 3 The results of the analysis of physico-chemical parameters of sodium alginate Indicators FS requirements 42-3383-97 Sodium alginate obtained by complex processing of raw materials F. vesiculosus F. dictichus F. serratus A. nodosum Description Amorphous slightly cream-colored or light gray with a creamy tint powder, odorless Amorphous slightly cream powder, odorless Solubility Slowly soluble in water with the formation of turbid colloidal solutions, practically insoluble in 95% alcohol, in chloroform and ether (GP XI, issue 1, p.175) Acc. Acc. Acc. Acc. Authenticity Qualitative reaction with a 2.0% solution of calcium chloride with the formation of a gel-like precipitate. The reaction is positive. The reaction is positive. The reaction is positive. The reaction is positive. Qualitative reaction with hydrochloric acid diluted to form a gel-like precipitate. The reaction is positive. The reaction is positive. The reaction is positive. The reaction is positive. Qualitative reaction with a saturated solution of ammonium sulfate, no precipitate should form. The reaction is positive. The reaction is positive. The reaction is positive. The reaction is positive. Sodium salt moistened with hydrochloric acid and introduced into a colorless flame turns it yellow. (GF XI, issue 1, p. 162) The reaction is positive. The reaction is positive. The reaction is positive. The reaction is positive. Substances insoluble in boiling water No more than 0.2% 0.11% 0.07% 0.15% 0.19% Mass loss on drying No more than 20.0% 13.5% 15.2% 12.6% 9.8% Sulphated Ash and Heavy Not more than 35.0% 19.8% 9.5% 14.5% 18.9% metals No more than 0,001% Acc. Acc. Acc. Acc. Arsenic No more than 0,0002% Acc. Acc. Acc. Acc. Kolich. defined Not less than 90.8% 96.1% 95.3% 97.0% 91.2%

Table 4 Examples of complex processing of dry thalli fucus algae by the present method. Raw materials Technological Modes The yield of the target product,% relative to raw materials one 2 3 F. vesiculosus Example 1. The raw material was extracted with azeotrope methylene chloride - ethanol in a Soxhlet apparatus for 5 hours. The extract was concentrated, dried. Algae meal was extracted by circulating percolation with ethanol 85% at a temperature of 65 ° C, infusion time of 5 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0 ÷ + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 5% aqueous ethanol, pH = 1-2. The infusion time is 6 hours, the temperature is 60 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Dried in a vacuum dryer. The meal was extracted by percolation with a 1.5% sodium carbonate solution at a temperature of 55 ± 5 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven. TIC - 3.10% Mannitol - 9.04% Fucoidan-P - 21.30% Sodium alginate - 30.05% Example 2. The raw material was extracted with azeotrope methylene chloride - ethanol in a Soxhlet apparatus for 8 hours. The extract was concentrated, dried. Algae meal was extracted by circulating percolation with ethanol 95% at a temperature of 45 ° С, infusion time was 3 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0 ÷ + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 15% aqueous ethanol, pH = 1-2. The infusion time is 9 hours, the temperature is 40 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Dried in a vacuum dryer. Meal was extracted by percolation with a 2.5% sodium carbonate solution at a temperature of 45 ± 5 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven. Timber industry - 3.22% Mannitol - 8.85% Fucoidan-P - 24.10% Sodium alginate - 27.12% F. distichus Example 3. The raw material was extracted with the chloroform - ethanol azeotrope in a Soxhlet apparatus for 6 hours. The extract was concentrated, dried. Algae meal was extracted by percolation with ethanol 85% at a temperature of 65 ° C, the infusion time was 5 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0- + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 10% aqueous ethanol, pH = 1-2. The infusion time is 6 hours, the temperature is 70 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Dried in a vacuum dryer. Meal was extracted by percolation with a 1.5% sodium carbonate solution at a temperature of 45 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven. TIC - 3.56% Mannitol - 10.85% Fucoidan-D - 18.75% Sodium Alginate - 27.06% Example 4. The raw material was extracted with the chloroform - ethanol azeotrope in a Soxhlet apparatus for 8 hours. The extract was concentrated, dried. Algae meal was extracted by percolation with ethanol 95% at a temperature of 45 ° C, the infusion time was 7 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0 ÷ + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 20% aqueous ethanol, pH = 1.5. The infusion time is 9 hours, the temperature is 50 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Dried in a vacuum dryer. The meal was extracted by percolation with a 1.5% sodium carbonate solution at a temperature of 55 ± 5 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven. TIC - 2.95% Mannitol - 9.87% Fucoidan-D - 15.47% Sodium alginate - 22.95% F. serratus Example 5. The raw material was extracted with azeotrope methylene chloride - ethanol in a Soxhlet apparatus for 8 hours. The extract was concentrated, dried. Algae meal was extracted by percolation with ethanol 90% at a temperature of 60 ° C, infusion time was 8 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0 ÷ + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 20% aqueous ethanol, pH = 3. The infusion time is 9 hours, the temperature is 75 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Freeze-dried. Meal was extracted by percolation with a 2% sodium carbonate solution at a temperature of 60 ± 5 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Freeze-dried. TIC - 1.85% Mannitol - 8.73% Fucoidan-S - 20.54% Sodium alginate - 18.90% Example 6. The raw material was extracted with azeotrope methylene chloride - ethanol in a Soxhlet apparatus for 8 hours. The extract was concentrated, dried. Algae meal was extracted by percolation with ethanol 95% at a temperature of 50 ° C, infusion time was 8 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0 ÷ + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 30% aqueous ethanol, pH = 4. The infusion time is 12 hours, the temperature is 65 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Freeze-dried. Meal was extracted by percolation with a 4% sodium carbonate solution at a temperature of 50 ± 5 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Freeze-dried. LPC - 2.60% Mannitol - 7.95% Fucoidan-S -17.91% Sodium Alginate - 22.27% A. nodosum Example 7. The raw material was extracted by percolation at a temperature of 20 ± 2 ° C with a mixture of chloroform - ethanol at a ratio of 1: 1, the infusion time was 24 hours. The extract was concentrated, dried. Algae meal was extracted by percolation with ethanol 85% at a temperature of 75 ° C, infusion time 4 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0 ÷ + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 5% aqueous ethanol, pH = 4. The infusion time is 12 hours, the temperature is 70 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Dried in a vacuum dryer. Meal was extracted by percolation with a 1.5% sodium carbonate solution at a temperature of 65 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven. TIC - 1.30% Mannitol - 5.05% Fucoidan-A - 11.60% Sodium alginate - 10.89% Example 8. The raw material was extracted by percolation at a temperature of 20 ± 2 ° C with a mixture of chloroform - ethanol at a ratio of 1: 1, the infusion time was 20 hours. The extract was concentrated, dried. The algae meal was extracted by percolation with ethanol 95% at a temperature of 65 ° C, the infusion time was 4 hours. The extract was filtered, concentrated in vacuo on a rotary evaporator. Crystallized at 0 ÷ + 4 ° C for 24 hours. Mannitol was filtered on a suction filter and dried, purified by column chromatography using alumina. Algae meal was extracted by circulating percolation with 15% aqueous ethanol, pH = 5.5-6. The infusion time is 18 hours, the temperature is 80 ° C. The extract was filtered, concentrated on a rotary evaporator. Conc. the extract was dialyzed in water purified through a cellophane membrane. Dried in a vacuum dryer. Meal was extracted by percolation with a 2.5% sodium carbonate solution at a temperature of 75 ° C. The extract was treated with sulfuric acid, the precipitate was dissolved in a 1.5% sodium carbonate solution. The sodium alginate solution was concentrated on a rotary evaporator, purified from ballast substances by dialysis in water purified through a cellophane membrane. Dried in a vacuum oven. LPK - 1.0% Mannitol - 4.41% Fucoidan-A - 14.00% Sodium Alginate - 13.66%

Claims (5)

1. The method of complex processing of Fucus vesiculosus Fucus, which consists in crushing dry algae thalli, extracting with a mixture of organic extractants, concentrating in a vacuum on a rotary evaporator, drying in a vacuum on a rotary evaporator, drying in a vacuum oven to obtain a lipid-pigment of the complex - thick extract, then the meal is extracted by the method of circulation percolation, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried and purified by column chromatography using alumina and purified mannitol is obtained, then the remaining meal is extracted by circulating percolation with 5-15% aqueous ethanol, concentrated in a vacuum on a rotary evaporator, dried in a vacuum oven, and a polysaccharide complex is obtained - dry Fucus extract, then the meal is extracted with a 1.5% solution of sodium carbonate, treated with sulfuric acid, concentrated on a rotary evaporator, purified by dialysis through a cellophane membrane, dried in a vacuum oven or lyophilization and it produces a sodium alginate polysaccharide, characterized in that in order to obtain a lipid-pigment complex (thick extract of fucus), extraction is carried out with a mixture of methylene chloride and ethyl alcohol in a ratio of 94.2: 5.86% in a Soxhlet apparatus, to obtain algae meal, the algal meal is extracted by the method percolation with 85-90% ethanol with forced circulation of the extractant after each hour of infusion for 3-5 hours, to obtain a dry extract of Fucus, meal was extracted with 5-15% ethanol solution at pH 1-2 and the infusion time I am 6-9 hours at a temperature of 40-60 ° C. 2. The method of complex processing of Fucus algae species of Fucus serratus (Fucus serratus) or double-sided fucus (Fucus distichus), which consists in that dry thalli of algae are crushed, extracted with a mixture of organic extractants, concentrated in a vacuum on a rotary evaporator, dried in a vacuum drying oven to obtain a lipid-pigment complex - a thick extract, then subsequently extracted with ethanol, a water-alcohol mixture and sodium carbonate to obtain mannitol, a polysaccharide complex and sodium alginate, excellent characterized in that to obtain a lipid-pigment complex (thick extract of Fucus), the extraction is carried out with an azeotropic mixture of methylene chloride and ethyl alcohol in a ratio of 94.2: 5.86% or an azeotropic mixture of chloroform with ethanol in a ratio of 88.06: 11.9% in Soxhlet apparatus, then the algal meal is extracted by percolation with 85-90% ethanol at a temperature of 45-65 ° C, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried, purified by column chromatography using alumina and purified mannitol is obtained, then the remaining meal is extracted by circulating percolation with 10-30% aqueous ethanol at pH 1-4, the infusion time is 6-12 hours at a temperature of 45-75 ° C, and it is purified by vacuum concentration followed by dialysis through cellophane membrane or by concentration on an ultrafiltration apparatus with simultaneous dialysis, dried by lyophilization or in a vacuum oven and get a polysaccharide complex - a dry extract of Fucus, then the meal is extracted by percolation with a 1.5-4% carb solution sodium onate at a temperature of 45-65 ° C, the extract is treated with sulfuric acid, the resulting alginic acid precipitate is dissolved in a 1.5% sodium carbonate solution, concentrated on a rotary evaporator, purified by dialysis through a cellophane membrane, dried in a vacuum oven or lyophilization and get polysaccharide sodium alginate. 3. The method according to claim 2, characterized in that to obtain a dry extract of fucus from fucus bilateral (Fucus distichus), the meal was extracted with a 10-20% solution of ethanol at pH 1-2 and the infusion time of 6-9 hours at a temperature of 50- 70 ° C. 4. The method according to claim 2, characterized in that to obtain a dry extract of Fucus from Fucus serratus (Fucus serratus), the meal was extracted with a 20-30% solution of ethanol at pH 2-4 and the infusion time of 9-12 hours at a temperature of 65- 75 ° C. 5. A method for the complex processing of Fucus algae of the Ascophyllum nodosum type (Ascophyllum nodosum), which consists in crushing dry algae thalli, extracting it with a mixture of organic extractants, concentrating in a vacuum on a rotary evaporator, drying in a vacuum on a rotary evaporator, and drying in a vacuum oven. to obtain a lipid-pigment complex - a thick extract of ascofillum, then subsequently extracted with ethanol, a water-alcohol mixture and sodium carbonate to obtain mannitol, a dry extract and alginate n atria, characterized in that to obtain a lipid-pigment complex (thick ascofillum extract), the extraction is carried out with a mixture of chloroform and ethanol in a 1: 1 ratio by percolation with forced circulation of the extractant after every hour of infusion for 20-24 hours, then the algal meal is extracted by the method percolation with 85-95% ethanol at a temperature of 65-75 ° C, filtered, concentrated in vacuo on a rotary evaporator, crystallized, filtered on a suction filter, dried, purified by column chromatography using Alumina is formed and purified mannitol is obtained, then the remaining meal is extracted by circulating percolation with 5-15% aqueous ethanol at pH 4-6, the infusion time is 12-18 hours at a temperature of 70-80 ° C, the extract is filtered, concentrated in vacuo on rotary evaporator, dialyzed in purified water through a cellophane membrane, dried by lyophilization or in a vacuum oven and get a polysaccharide complex - dry ascofillum extract, then the meal is extracted by percolation with a 1.5-2.5% sodium carbonate solution at temperatures e 65-75 ° C, the extract is treated with sulfuric acid, the resulting alginic acid precipitate is dissolved in a 1.5% sodium carbonate solution, concentrated on a rotary evaporator, purified by dialysis in purified water through a cellophane membrane, dried in a vacuum oven or lyophilization and get polysaccharide sodium alginate.

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