RU2708157C1 - Method of producing carotenoids from hydrobionts - Google Patents

Abstract

FIELD: fishing and fish farming.

SUBSTANCE: invention relates to fish industry, in particular, to methods for extracting fat-soluble carotenoids. Method of producing carotenoids from hydrobionts involves preparation of raw material, its extraction with a polar organic solvent, filtration of the extract, connection with oil and re-extraction in the darkness, separation of the oil extract, washing with water and drying. Hydrobionts are represented by anadara broughtoni and/or Mactra chinensis, the extracting agent used is 50 % propylene glycol containing citric acid and ascorbic acid in ratio of 1,000:0.5:0.5, respectively. Extraction is performed for 12 hours at raw material:extractant ratio of 1:3–4. Oil is represented by a mixture of soybean oil with sunflower or corn oil at ratio of 57–60:43–40, respectively. Ratio of extract:mixture of oils is 1:1–1.5. Re-extraction is carried out for 12 hours at temperature of 30–35 °C.

EFFECT: invention increases efficiency of extracting carotenoids from hydrobionts, reduces time of extraction and re-extraction, as well as increases stability of carotenoids during production and storage.

1 cl, 4 tbl, 4 ex

Description

The invention relates to the fishing industry, in particular to methods for the isolation of fat-soluble carotenoids, and can be used to obtain a product of vegetable fat of a balanced fatty acid composition with a complex of carotenoids, which can be used in medicine as food additives with therapeutic and preventive properties, cosmetology, in food industries for the preparation of fish products, mayonnaise, sauces, in aquaculture.

A number of inventions are known, involving the use and / or preparation of extracts of biologically active compounds: saponins, phospholipids, carotenoids from marine hydrobionts, such as cucumaria, sea urchins, etc., for their subsequent enrichment of fish oil (RF patents Nos. 2129809, 2215532 , 2112527, 2181976, 2050388, application for invention of the Russian Federation No. 97106885/13).

In accordance with the above inventions, extracts of biologically active substances (BAS) are obtained by repeated extraction of the raw materials with organic solvents, such as ethanol, acetone, alcohols, ethers.

This process of enriching fat with biologically active substances (including fat-soluble ones) is rather laborious and expensive, requires the use of an additional solvent, when extracting biologically active substances, most of the native qualities of raw materials are lost, because of the use of chemical solvents the method is not environmentally friendly.

There are other ways to enrich fats with fat-soluble biologically active substances.

A known method of producing carotenoids by extracting soybean oil from crustaceans to use astaxanthin-enriched oil as additives in the salmon diet to accelerate the pigmentation process (Extraction of astaxanthin using soybean oil from waste generated during the processing of crustaceans (lobsters). / Chen HM, Meyers SP ; TSNIITEIRKh.Lane No. 6729. - 20 C. - 8. - J. Food Sci., 1982, v. 47, No. 3, p. 892-896).

A known method of producing carotenoids from waste obtained during processing of cephalopods, including oil extraction and heat treatment of the feedstock (RF patent No. 2064476, published July 27, 1996).

The method involves the use of liver, gonads, residues of the gastrointestinal tract or whole cephalopods as raw materials, and grinding the raw material to a particle size of 1-3 mm.

The ground mass is subjected to heat treatment in two stages to prevent protein coagulation: 1 - at a temperature of 18-20 ° C to 50-60 ° C; 2 - from 18-20 ° C to 50-60 ° C, then - exposure to a temperature of 80-85 ° C for 30-50 minutes. As a result of heating, the protein contained in it coagulates and the homogeneous mass is divided into a dense fraction and a liquid fraction containing carotenoids dissolved in lipids.

At this stage, the process of dissolution of carotenoids in lipids of the raw material is carried out. After heat treatment, the resulting mass by means of known methods is divided into liquid and dense fractions. The selection of carotenoids is carried out from the liquid fraction by any of the known methods: separation or sedimentation. The yield of dye from 100 kg of raw material was 16-30 kg during separation, with sedimentation 5-10 kg (depending on the fat content of the waste). The dye can be used as an enrichment of biologically active substances.

A known method of producing fish oil with carotenoids, which involves grinding the raw material on a large grid (20 mm), its heat treatment in several stages: first at a temperature of 60 to 85 ° C for 1 hour, then for 20 minutes at 80-90 ° C and then conducting separation at a temperature not lower than 50 ° C of the fat-containing fraction in the collector-heater, and then separation at a temperature of 80 to 90 ° C. The finished product is fish oil containing natural phospholipids and carotenoids (application for invention of the Russian Federation No.2000123494 / 14, date of publication of the application September 27, 2003).

To obtain encapsulated fish oil with biologically active phospholipids and carotenoids in their native form, low-value raw materials were used: capelin capelin and herring, which contain a large amount of fat, i.e. raw lipids are used as a solvent for the extraction of carotenoids.

The disadvantages of the method include the multi-stage and duration of the process.

The inventions considered above provide for the use of cephalopods only, some crustaceans and / or their processing waste as raw materials.

As the closest analogue, the method of obtaining a complex of fat-soluble carotenoids from ascidia was selected (RF patent No. 2339387, published November 27, 2008), including cutting ascidia, washing the obtained raw materials, grinding, drying, extraction with a polar organic solvent and packaging, according to the invention, as a raw material use raw or dried mantle or tunic or mantle and tunic from ascidia at a ratio of mantle and tunic 1: 2, and the raw material is dried at 40 ° C to a moisture content of 7-10%, extraction is carried out with 96% ethanol or acetone in in terms of raw materials: extractant 1: 1-1: 5 for 16-24 hours, the resulting extract is filtered, combined with oil and water in a ratio of 5: 1: 10 for raw materials and 2: 1: 4 for dry materials, respectively, in the dark reextraction is carried out for 22-24 hours, the oil extract is separated, washed with water, and then the finished product is dried.

The disadvantages of the closest analogue are the insufficiently high concentration of carotenoids in the extract, a long extraction time, the instability of carotenoids in the process of obtaining and storage due to oxidation.

The problem to which the invention is directed is the development of a method that provides an increase in the content of carotenoids and biological effectiveness of the finished product, a reduction in the time of extraction and re-extraction, an increase in the stability of carotenoids in the process of obtaining and storage.

The technical result that manifests itself in solving the problem is to increase the efficiency of extraction of carotenoids from hydrobionts, reduce the time of extraction and re-extraction, increase the stability of carotenoids in the process of obtaining and storage, and reduce the complexity.

The problem is solved in that in the method for producing carotenoids from aquatic organisms, including the preparation of raw materials, extraction with a polar organic solvent, the resulting extract is filtered, combined with oil and re-extracted in the dark, the oil extract is separated, washed with water and then the finished product is dried , as hydrobionts use Browton's anadara and / or Maktra Chinese, as extractant use 50% propylene glycol containing citric acid and ascorbic acid in a ratio of 1000: 0.5 : 0.5, respectively, extraction is carried out for 12 hours at a ratio of raw materials: extractant 1: 3-4, the resulting extract is combined with a mixture of soybean oil with sunflower or corn at a ratio of 57-60: 43-40, respectively, and the ratio of extract: mixture oils is 1: 1-1.5, and reextraction is carried out for 12 hours at a temperature of 30-35 ° C.

A comparative analysis of the set of essential features of the proposed technical solution and the set of essential features of the prototype and analogues indicates its compliance with the criterion of "novelty".

In this case, the distinguishing features of the claims solve the following functional tasks.

The signs “use Browton's anadara and / or Chinese maktru as hydrobionts” allow expanding the raw material base for obtaining enriched fats.

The sign “50% propylene glycol is used as an extractant” helps to increase the efficiency of extraction of carotenoids from the feedstock, in addition, propylene glycol is highly soluble in water, approved for use in the food industry and is a dihydric alcohol.

The sign “use propylene glycol containing citric acid” also provides an increase in the efficiency of extraction of carotenoids from the feedstock, since the organic acid contributes to the destruction of carotenoprotein complexes.

It has been experimentally proven that citric acid increases the yield of carotenoids compared to malic and succinic acids.

The sign "use propylene glycol containing ascorbic acid" ensures the stability of carotenoids in the process of obtaining and storage, because ascorbic acid acts as a food antioxidant.

The sign “as extractant use 50% propylene glycol containing citric acid and ascorbic acid in a ratio of 1000: 0.5: 0.5, respectively” describes the composition of the extractant.

The signs "extraction is carried out for 12 hours at a ratio of raw materials: extractant 1: 3-4" describe the optimal operating characteristics of the extraction.

The sign "the obtained extract is combined with the mixture [oils], and the ratio of the extract: the mixture of oils is 1: 1-1.5" ensures the completeness of re-extraction and a high concentration of carotenoids in the finished product.

The sign "the extract is combined with a mixture of soybean oil with sunflower or corn at a ratio of 57-60: 43-40, respectively" provide a balanced fatty acid composition of the finished product.

The signs "reextraction is carried out for 12 hours at a temperature of 30-35 ° C" describe the optimal operating characteristics of the reextraction, moderate temperature allows you to maximize the properties and stability of carotenoids.

The inventive method is carried out according to standard technology on standard equipment.

As raw materials, soft tissues (muscle, mantle and viscera) of bivalve mollusks are used - anadara of Broughton and Maktra of China.

As raw materials can be used and waste obtained during their processing. It is possible to use other marine aquatic organisms that accumulate carotenoids in the metabolic process.

Prepared raw materials are ground to a particle size of 0.1 to 0.5 mm, then further homogenized.

Since carotenoids are photosensitive compounds, pieces of dry ice were additionally introduced at the stage of grinding the raw material and its homogenization to reduce the action of atmospheric oxygen and lower the temperature, which rises during mechanical action.

The crushed raw materials are placed in a glass container and filled with an extractant, which is used as 50% propylene glycol containing citric acid and ascorbic acid in a ratio of 1000: 0.5: 0.5, respectively, with a ratio of raw materials: extractant 1: 3-4 and hermetically sealed container to prevent contact with oxygen in the air to avoid oxidation of carotenoids.

The extraction is carried out by infusion at a temperature of 25-27 ° C for 12 hours.

The resulting extract is filtered through a gas cloth or centrifuged, placed in a separatory funnel, a mixture of soybean oil with sunflower or corn is added to it at a ratio of 57-60: 43-40, respectively, and the ratio of extract: oil mixture is 1: 1-1.5.

Reextraction is carried out in a dark place for 12 hours at a constant temperature of 30-35 ° C. After its passage, the oil extract is washed with water with vigorous mechanical stirring or shaking to remove traces of alcohol, and then dried in a known manner (filtration, centrifugation).

A dense residue that contains proteins can be used as a raw material to obtain a biologically active additive (BAA), for example, a source of free amino acids.

The target product is vegetable fat enriched with a complex of marine carotenoids.

The finished product in the form of an oil extract is a clear oily liquid (color from pinkish-orange to red) containing a complex mixture of carotenoids in an amount of 0.11-0.14 mg / ml, including fucoxanthin 3'-ether, fucoxanthinol 3'-ether , fucoxanthin, fucoxanthinol, etc.

The absence of impurities allows you to enter the resulting oil extract, for example, in the fat phase of mayonnaise and mayonnaise sauces without changing the organoleptic characteristics of the finished product.

Table 1

Organoleptic characteristics of oil extracts containing carotenoids

Index Description Appearance Oily liquid Colour Pinkish orange to red Taste and smell Inherent, without extraneous taste and smell Transparency at a temperature not above 60 ° C Transparent The presence of impurities Not allowed

Physico-chemical quality indicators of oil extracts containing carotenoids are presented in table 2.

table 2

Physico-chemical characteristics of oil extracts containing carotenoids

Index Indicator value The total content of carotenoids, mg / ml, not less 0.10 Mass fraction of water and non-greasy impurities,%, no more 0.5 Mass fraction of fat,%, not less than 99.0

The safety indicators of oil extracts containing carotenoids are presented in table 3.

Table 3

Safety Indicators for Oil Extracts Containing Carotenoids

Index Permissible Content Level Actual value Toxic elements, mg / kg no more Lead 0.3 0.005 Arsenic 0.1 0.009 Cadmium 0.05 not upd. Mercury 0.05 not upd. Pesticides mg / kg no more HCH (α-, β- and γ-isomers) 0.05 not upd. DDT, DDD, DDE 0.1 not upd. Polychlorinated biphenyls, mg / kg 3.0 0.06 Radionuclides, Bq / kg (l) no more Cesium-137 60 not upd. Strontium-90 80 not upd. Indicators of oxidative spoilage, meq / kg no more Peroxide value 10.0 4.1

The results of a comprehensive assessment of the antiradical properties of the obtained oil extracts of carotenoids from hydrobionts are presented in table 4.

Table 4

Antiradical activity of extracts from aquatic organisms

As can be seen from table 4, the extracts obtained using 50% propylene glycol have a higher antiradical activity than all others.

Examples of the method

Example 1

The raw frozen mantle of Mactra Chinese bivalve mollusk (Mactra chinensis) weighing 1 kg was thawed in air at a temperature of no higher than 20 ° C until the blocks disintegrated. Washed in running water, with a temperature not exceeding 20 ° C, placed on a sieve to remove excess water. Runoff duration 20-30 min. The prepared raw materials were crushed in an electric cutter to a homogeneous mass with a particle size of 0.1 to 0.5 mm, then further homogenized. Dry ice was used during the grinding and homogenization process.

The crushed raw materials are placed in a glass container, filled with a mixture of 3 l of 50% propylene glycol, 0.0015 kg of edible citric acid and 0.0015 kg of ascorbic acid, and the container is hermetically sealed to prevent contact with atmospheric oxygen. The extraction is carried out by infusion at 25-27 ° C for 12 hours

The resulting extract is filtered through gas tissue. The extract (3 L) is placed in a separatory funnel, 3 L of a mixture of vegetable oils (40% sunflower and 60% soybean) are added. Reextraction is carried out in a dark place for 12 hours at a temperature of 30 ° C. After its passage, the oil extract is drained, washed with water with constant stirring or shaking to remove traces of propylene glycol.

The final product - the oil extract is dried. The yield of the final product is 2.95 L per 1 kg of raw material. The concentration of carotenoids in the oil extract is 0.14 mg / ml.

Example 2

The processing of the raw-frozen insides of the Chinese mactra was carried out analogously to example 1. The prepared raw materials were crushed in an electric cutter to a homogeneous mass with a particle size of 0.1 to 0.5 mm, then further homogenized. Dry ice was used during the grinding and homogenization process.

The crushed raw material is placed in a glass container, filled with a mixture of 4 l of 50% propylene glycol, 0.0020 kg of edible citric acid and 0.0020 kg of ascorbic acid, and the container is hermetically sealed to prevent contact with atmospheric oxygen. The extraction is carried out by infusion at 25 ° C for 12 hours

The resulting extract is filtered through gas tissue. The extract (4 L) is placed in a separatory funnel, 6 L of a mixture of vegetable oils (43% corn and 57% soy) are added. Reextraction is carried out in a dark place for 12 hours at a temperature of 35 ° C. After its passage, the oil extract is drained, washed with water with constant stirring or shaking to remove traces of propylene glycol. The final product - the oil extract is dried.

The yield of the final product is 5.82 liters per 1 kg of raw material. The concentration of carotenoids in the oil extract is 0.11 mg / ml.

Example 3

The processing of raw materials and all operations were carried out analogously to example 1, but the mantle of Anadara Broughton (Anadara colletoni) was used as raw material. The crushed raw materials are placed in a glass container, filled with a mixture of 3 l of 50% propylene glycol, 0.0015 kg of edible citric acid and 0.0015 kg of ascorbic acid, and the container is hermetically sealed to prevent contact with atmospheric oxygen. The extraction is carried out by infusion at 25 ° C for 12 hours

The resulting extract is filtered through gas tissue. The extract (3 L) is placed in a separatory funnel, 3 L of a mixture of vegetable oils (40% sunflower and 60% soybean) are added. Reextraction is carried out in a dark place for 12 hours at a temperature of 35 ° C. After its passage, the oil extract is drained, washed with water with constant stirring or shaking to remove traces of propylene glycol. The final product - the oil extract is dried.

Received 2.84 kg of oil extract per 1 kg of raw materials with a concentration of carotenoids of 0.12 mg / ml

Example 4

The processing of raw materials was carried out analogously to example 1, but as the raw material used all the soft tissue of the Chinese maktra and Broughton anadara in the ratio 1: 1.

The crushed raw materials are placed in a glass container, filled with a mixture of 3 l of 50% propylene glycol, 0.0015 kg of edible citric acid and 0.0015 kg of ascorbic acid, and the container is hermetically sealed to prevent contact with atmospheric oxygen. The extraction is carried out by infusion at 27 ° C for 12 hours

The resulting extract is filtered through gas tissue. The extract (3 L) is placed in a separatory funnel, 4.5 L of a mixture of vegetable oils (43% corn and 57% soy) are added. Reextraction is carried out in a dark place for 12 hours at a temperature of 35 ° C. After its passage, the oil extract is drained, washed with water with constant stirring or shaking to remove traces of propylene glycol. The final product - the oil extract is dried.

The yield of the final product is 4.35 L per 1 kg of raw material. The concentration of carotenoids in the oil extract is 0.13 mg / ml.

The advantages of the proposed method are:

- the use in the process of processing only natural products without the use of hazardous organic solvents allows you to preserve the native qualities of the hydrobiont as much as possible;

- obtaining a product with a high content of carotenoids due to the combination of extraction and reextraction at a moderate temperature regime, as well as through the use of citric acid as a destructive agent for carotene protein complexes;

- increasing the storage stability of the resulting oil extract from aquatic organisms through the use of ascorbic acid as a food antioxidant;

- increasing the biological effectiveness of the finished product and its balanced fatty acid composition due to the use of a mixture of vegetable oils as a stripping agent;

- reducing the complexity - the method is simplified, waste-free, its implementation does not require large energy costs, the use of complex and expensive equipment;

The resulting products can be successfully used as food additives with therapeutic and prophylactic properties for fat-containing food products, as well as additives in the diet of fish, in particular salmon, to accelerate the pigmentation process.

Claims (1)

A method of producing carotenoids from aquatic organisms, including the preparation of raw materials, extraction with a polar organic solvent, the resulting extract is filtered, combined with oil and re-extracted in the dark, the oil extract is separated, washed with water and then the finished product is dried, characterized in that as hydrobionts use Browton's anadara and / or Maktra Chinese, 50% propylene glycol containing citric acid and ascorbic acid in a ratio of 1000: 0.5: 0.5, respectively, are used as extractant extraction is carried out for 12 hours at a ratio of raw materials: extractant 1: 3-4, the resulting extract is combined with a mixture of soybean oil with sunflower or corn at a ratio of 57-60: 43-40, respectively, and the ratio of extract: oil mixture is 1: 1- 1.5 and re-extraction is carried out for 12 hours at a temperature of 30-35 ° C.