RU2476211C1 - Method for producing alkyl-glycerol esters of sea fats - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to pharmaceutical and food industry, namely a method for producing alkyl-glycerol esters of sea fats. The method for producing alkyl-glycerol esters of sea fats involving hydrolysis of sea fat, acidation of a fatty mixture, water washing, first crystallisation of the mixture of acetone and hexane, filtration, second crystallisation of the residue of acetone or hexane, filtration and drying of an end product under certain conditions.

EFFECT: method enables reducing a time of technological process.

2 cl, 21 ex

Description

The invention relates to the food and pharmaceutical industries and can be used to produce alkyl-glycerol esters with high biological effect from sea fats.

Under the name "sea lipids" a large number of biologically active substances are combined - polyunsaturated fatty acids, phospholipids, hydrocarbons, such as squalene and squalane, phytosterols and others. The total lipid content of some hydrobionts includes unusual lipids - 1-O-alkyl-diacylglycerols - compounds formed by fatty acids and alcohols (butyl, chymil and selachilic, etc. - 1-O-alkyl-glycerol ethers (AGE)).

Alkyl radicals in the 1st position of glycerol are saturated, both in the case of chymyl and batyl alcohols, and unsaturated in selachyl alcohol. Usually in natural mixtures, the sum of these 3 alcohols is more than 90% of all AGE. To release AGE and their further concentration, the ester bonds between glycerol and fatty acids in the 2nd and 3rd positions are hydrolyzed.

Ether-bound lipids, AGEs, have been involved in the formation of the mammalian immune system from the very first days of life and are one of the main factors supporting its normal functioning throughout the entire period. Their effect on hematopoiesis is widely known, and, according to researchers, AGE normalize bone marrow function without causing excessive stimulation. In this regard, AGE is successfully used in rehabilitation therapy in the treatment of cancer, radiation injuries, immunodeficiency states, etc. (Brohult A., Brohult J., Brohult S., Joelsson I. Reduce mortality in cancer patient after administration of alkylglycerols // Acta Obst. Gynecol. Scand. 1986. Vol.65 P.779-785; Miter R., Etienne M., Martinais S., Salmon H., Allaume P., Legrand P., Legrand AB Humoral defense improvement and haematopoiesis stimulation in sows and offspring by oral supply of shark-liver oil to mother during gestation and lactation // British J. Nutr. 2005. Vol. 94. P.753-762).

At the moment, there is no way that would, while maintaining the efficiency of the process (yield and purity of the product) drastically reduce its duration, thereby intensifying the process and giving manufacturing enterprises the possibility of more processing of procured raw materials and, consequently, production of finished products.

A known method for the isolation and concentration of alkyl glycerol esters from marine fats (Patent GB 1,076,706 A process for the preparation and concentration of free hydroxylic substances from marine oils, 08/17/1965, Int. Cl .: C07C 43/02). The method consists in the following: sea fat (shark liver oil) is transesterified with methyl alcohol (ethyl and isopropyl allowed) are converted to fatty acid methyl esters, while alkyl glycerol esters are released into the mixture, which are extracted from the total mixture with water-methanol or water-water ethanol solutions, then evaporated, obtaining the target AGE.

The disadvantages of the method:

1) the use of methanol is unacceptable in food production and is not desirable in pharmaceutical;

2) at the first stage of obtaining fatty acid esters, the use of anhydrous sea fat and alcohol is necessary, otherwise the reaction will not go through to the end and all subsequent manipulations are useless; for fat, this turns into an additional drying stage, which requires an apparatus with a stirrer, steam heating, vacuum, which is problematic in non-specialized industries, and for alcohols this is more or less solved only with methanol, for ethyl and isopropyl alcohols, complete dehydration requires chemical treatment metal calcium or magnesium chips, which is unsafe and requires a certain professionalism;

3) the method involves the use of a well-established technological line, which complicates its implementation in small and other fishery and food enterprises of the domestic industry;

4) using this method, it is possible to obtain AGE with a purity of only 23-75%, which is insufficient for commercial use and suggests further additional stages of purification;

5) the method is quite laborious and, judging by the description, requires increased economic costs for its functioning.

A known method for the isolation of alkyl glycerol esters from squid squid fat Berryteuthis magister (Hayashi K / Isolation of alkyl glyceryl ethers from liver oil unsaponifiables by recrystallization // Bulletin of the Japanese Society of Scientific Fisheries. Vol.52, N8. 1986. P.1475 ) The method consists in the fact that the fat extracted from squid liver is hydrolyzed with potassium alkali, the unsaponifiable substances (and AGE in their composition) are extracted with diethyl ether, the diethyl ether is evaporated and the unsaponifiables are crystallized from hexane at a temperature of 2 ° C, obtaining a precipitate of alkyl glycerol ethers, which are then filtered.

The disadvantages of the method are:

1) the use of diethyl ether in technology, which limits the use of the resulting preparation in the food industry and medicine;

2) the stage of extraction of unsaponifiable substances with diethyl ether is a rather complicated process in hardware design when it is scaled from laboratory flasks to production devices;

3) the emulsions formed at the extraction stage are extremely stable, which forces experimenters to use an extractant (diethyl ether) in excess to reduce the loss of extractable substance;

4) although the yield of AGE is 52.4%, and the purity of the product is 99.0%, it is impossible to recommend the use of this method in production due to significant amounts of diethyl ether, which transfers the process to the category of fire and explosion hazard.

A known method for the allocation of alkyl-glycerol esters from the fat of the squid squid (Popkov A.A., Kasyanov S.P., Ovchinnikov V.V., Akulin V.N. T.120, 1997, S.77-85). The method consists in the fact that the fat extracted from squid liver is hydrolyzed with potassium alkali, the reaction mixture is acidified, the separated free fatty acids and unsaponifiable substances are separated from the water-salt layer, washed with water and crystallized twice from hexane, at the beginning the fat mixture is dissolved in hexane at the ratio of the fat mixture: hexane = 1: 15, heating to 60 ° C, then cooled to 0 ° C and incubated for 24 hours, after which the precipitate (intermediate product) is separated by filtration, dried and a second crystal is carried out hexane at the same temperature and for only 2 hours, but with a ratio of intermediate product - hexane = 1: 20, kg / l.

The disadvantages of the method are:

1) not optimized ratios of the fat mixture: hexane during crystallization, which do not provide a sufficient yield of the product (47%) and its purity (97.0%);

2) the temperature of 0 ° С is chosen as the main crystallization temperature, which does not correspond to the true temperature range admissible for this process and a specific solvent;

3) the authors, despite the large amount of work, could not provide the required product yield and purity, did not bring any alternative solvent for the fractionation of AGE, which reduces the possibility of their production in enterprises and makes it dependent on only one solvent.

Closest to the claimed method is a method for the isolation of alkyl glycerol esters from the fat of Commander squid (RF Patent No. 2415125 of 03/27/2011). The method includes hydrolysis of sea fat to isolate free fatty acids and release an unsaponifiable fraction containing AGE, neutralizing the resulting fat mixture with acid, washing with water and sequential double crystallization from an organic solvent, filtering and drying the precipitate formed after each crystallization. For the first and second crystallization, the fat mixture in an organic solvent is kept first for 10-12 hours at room temperature, and then for 12-15 hours at 0-4 ° C.

Acetone or hexane is used in the process, and only one solvent can be used in the whole process, or a combination thereof.

When using hexane as an organic solvent, the ratio of fat mixture to hexane in the first crystallization is 1:10, kg / l, and in the second crystallization of the intermediate product 1:50, kg / l.

When using acetone as an organic solvent, the ratio of fat mixture to acetone is 1:10, kg / l, both in the first and second crystallization of the intermediate product.

The method allows you to get AGE with a yield of 50-59.0% and a purity of more than 99%.

Disadvantages:

1) the method uses significant volumes of solvents, which requires the use of large containers, appropriate filtering systems and continuous regeneration of solvents;

2) the two crystallizations of the method are separated by drying the precipitate so that when changing the solvent there is no contamination of one solvent with another; drying really interrupts the process and postpones the subsequent 2nd crystallization for several hours;

3) the duration of the entire process - from the initial fat to the target product (AGE) - is at least 59 hours, an average of 65 hours, despite the fact that the process between the stages is inextricable. Such a long process time has an extremely negative effect on the performance of the AGE production line and does not allow for quick and timely processing of incoming raw materials.

The problem solved by the invention is to reduce the time of the technological process for the production of alkyl glycerol esters (AGE) from sea fats due to the instant formation of crystals of the target product.

The problem is solved in that in the known method for producing alkyl-glycerol esters from sea fats, including hydrolysis of sea fat, acidification of the fat mixture, washing with water, first crystallization of the mixture with acetone or hexane, filtration, second crystallization of the precipitate obtained from acetone or hexane, filtering and drying the target product, while the first and second crystallization uses one of these solvents, according to the claimed method, the ratio of the fat mixture to acetone or hexane having perature from (-15 ° C) to (-5 ° C), the first and second crystallization is 1: 1-5 kg / l.

The precipitates formed in the stages of the first and second crystallization, after filtration, are washed with a small amount of acetone or hexane having a temperature from (0 ° C) to (+ 4 ° C), which allows to further increase the purity of the target product.

The ratio of the fat mixture: acetone or hexane (organic solvent) during the first crystallization is 1: 1-5, the second also 1: 1-5 (kg / l), which creates the conditions for almost instant formation of AGE crystals and, as a result, allows in contrast to the prototype, abandon the long crystallization process, which, in the final result, reduces the technological process by 5-6 times.

When an organic solvent is used in the technological process at the crystallization stages in an amount of less than one volume relative to the initial amount of the fat mixture obtained by hydrolysis, it leads to the formation of a very thick mixture, which not only complicates the crystallization of the target product, but also is difficult to filter. As a result of this, not only the time of the technological process increases, but also the yield and quality of the target product decreases due to the capture of fatty acids in the precipitate. The use of an organic solvent volume in excess of five times the initial amount of the fat mixture in the process is also undesirable, since a significant dilution of the mixture occurs, which leads not only to excessive consumption of the organic solvent, but also requires an increase in the time for filtering the mixture and its subsequent regeneration .

Processing the fat mixture for crystallization in both the first and the second with an organic solvent having a temperature from (-15 ° С) to (-5 ° С) creates optimal conditions for the crystallization of AGE from the mixture, with the formation of AGE crystals almost instantly, which leads to a significant reduction in the time to obtain the target product with a high yield and a high degree of purity.

The use for crystallization of an organic solvent having a temperature below (-15 ° C) leads to a deterioration in the quality of the target product due to the intense enrichment of the precipitate with ballast saturated fatty acids. And the use of an organic solvent having a temperature above (-5 ° C) also does not ensure the achievement of the claimed technical result, since crystals of the target product are poorly formed and only partially precipitate, the product is mainly in solution. This, in the end result, leads to a significant increase in the duration of the technological process, since in order to obtain the target product with the required yield, it will be necessary to conduct the crystallization process a second time.

The temperature of the organic solvent for washing the precipitates formed in the first and second stages of crystallization, from (0 ° C) to (+ 4 ° C) is the most optimal. Raising or lowering the temperature of the organic solvent for washing negatively affects the yield and purity of the target product.

It should be emphasized once again that acetone or hexane can be taken as organic solvents, and the entire process should use only one solvent. As a result, no additional time is required for the separation of solvents (cleaning one solvent from another). The duration of the process according to this method is, on average, 11-12 hours.

The work was carried out on the lipids from the liver of mass commercial objects, such as commander squid (up to 20% AGE) and Kamchatka crab (4% AGE).

The method is as follows.

EXAMPLE 1. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, sea fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in fat is 20% by weight.

Sea fat is placed in an enameled vessel with a capacity of 10 l with a stirrer and heating, a solution of potassium alkali (0.2 kg) in 2.0 l of 80% ethanol is added, and then the mixture is stirred for 2 hours at 50 ° C for completion of the hydrolysis reaction. After which the mixture is acidified with a 15% aqueous solution of sulfuric acid. The lower water-salt layer is removed, and the upper fat is washed three times with equal volumes of water heated to 50 ° C. Received 950 g of a mixture of fatty acids with AGE.

For the first crystallization, the mixture is placed in a vessel with a capacity of 5.0 l, 0.95 l of acetone is added, having a temperature of -15 ° C (based on the mixture-acetone ratio 1: 1, kg / l) with vigorous stirring, after which the precipitate is filtered off on the filter and washed with 0.5 l of acetone having a temperature of 0 ° C.

The precipitate obtained is redissolved in acetone, based on a ratio of 1: 1 (kg / l), with vigorous stirring and a second filtration is carried out, for which all operations from the previous stage are exactly repeated.

Then the precipitate is dried at room temperature to constant weight.

Received 0.104 kg of powder with an AGE content of 99.0% by weight.

The total yield of 51.4%.

The duration of the process was 11 hours.

EXAMPLE 2. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 1, except that the ratio of fat mixture to acetone during the second crystallization was 1: 5, kg / l, and the precipitate was washed with a temperature solvent of + 4 ° С.

Received 0.111 kg of powder with an AGE content of 99.2% by weight.

The total yield of 55.0%.

The duration of the process was 11 hours.

EXAMPLE 3. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 1, except that the ratio of fat mixture to acetone during the first crystallization was 1: 5, kg / l, and the precipitate was washed with acetone having a temperature of + 2 ° C.

Received 0.112 kg of powder with an AGE content of 99.2% by weight.

The total yield of 55.6%.

The duration of the process was 11.0 hours.

EXAMPLE 4. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 3, except that the ratio of fat mixture to acetone during the second crystallization was 1: 5, kg / l.

Received 0.119 kg of powder with an AGE content of 99.6% by weight.

The total yield of 59.2%.

The duration of the process was 12 hours.

EXAMPLE 5. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 1, except that the temperature of the organic solvent, acetone, when mixed was -5 ° C, and the precipitate was washed with a solvent of the same temperature

(-5 ° C).

Received 0.106 kg of powder with an AGE content of 99.0% by weight.

The total yield of 52.3%.

The duration of the process was 11 hours 45 minutes.

EXAMPLE 6. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 2, except that the temperature of the organic solvent, acetone, when mixed was -5 ° C.

Received 0.113 kg of powder with an AGE content of 99.1% by weight.

The total yield of 55.8%.

The duration of the process was 11 hours 10 minutes.

EXAMPLE 7. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 3, except that the temperature of the organic solvent, acetone, when mixed was -5 ° C.

Received 0.113 kg of powder with an AGE content of 99.2% by weight.

The total yield of 56.1%.

The duration of the process was 11 hours 5 minutes.

EXAMPLE 8. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 4, except that the temperature of the organic solvent, acetone, when mixed was -5 ° C.

Received 0.118 kg of powder with an AGE content of 99.4% by weight.

The total yield of 58.8%.

The duration of the process was 11 hours 44 minutes.

EXAMPLE 9. Isolation of AGE from Commander Squid Liver Fat when acetone is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 2, except that the temperature of the organic solvent, acetone, when mixed on the first crystallization was -15 ° C, and on the second crystallization -5 ° C.

Received 0.114 kg of powder with an AGE content of 99.3% by weight.

The total yield of 56.4%.

The duration of the process was 11 hours 198 minutes.

EXAMPLE 10. The allocation of AGE from the liver fat of the commander squid when using acetone in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 1, except that the ratio of the fat mixture and acetone in the first and second crystallizations was 1: 4, kg / l, and the temperature of the organic solvent, acetone, when mixed in the first crystallization was - 15 ° C, and the second -5 ° C.

Received 0.112 kg of powder with an AGE content of 99.1% by weight.

The total yield of 55.6%.

The duration of the process was 12 hours.

EXAMPLE 11. Isolation of AGE from Commander Squid Liver Fat when hexane is used in the crystallization process.

To obtain AGE, sea fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in fat is 20% by weight.

All operations in this example were carried out as in example 1, except that hexane was taken as an organic solvent.

Received 0.101 kg of powder with an AGE content of 99.1% by weight.

The total yield of 50.2%.

The duration of the process was 11 hours 30 minutes.

EXAMPLE 12. The allocation of AGE from the liver fat of the commander squid when used in the process of crystallization of hexane.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 2, except that hexane was taken as an organic solvent.

Received 0.103 kg of powder with an AGE content of 99.1% by weight.

The total yield of 51.1%.

The duration of the process was 11 hours 42 minutes.

EXAMPLE 13. Isolation of AGE from Commander Squid liver fat when using hexane in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 3, except that hexane was taken as an organic solvent.

Received 0.106 kg of powder with an AGE content of 99.3% by weight.

The total yield of 52.7%.

The duration of the process was 11 hours 25 minutes.

EXAMPLE 14. The allocation of AGE from the liver fat of the commander squid when used in the process of crystallization of hexane.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 4, except that hexane was taken as an organic solvent.

Received 0.117 kg of powder with an AGE content of 99.6% by weight.

The total yield of 58.4%.

The duration of the process was 11 hours 5 minutes.

EXAMPLE 15. Isolation of AGE from Commander Squid Liver Fat when hexane is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 5, except that hexane was taken as an organic solvent.

Received 0.104 kg of powder with an AGE content of 99.0% by weight.

The total yield of 51.5%.

The duration of the process was 11 hours 10 minutes.

EXAMPLE 16. Isolation of AGE from Commander Squid liver fat when using hexane in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 6, except that hexane was taken as an organic solvent.

Received 0.108 kg of powder with an AGE content of 99.3% by weight.

The total yield of 53.4%.

The duration of the process was 11 hours 55 minutes.

EXAMPLE 17. Isolation of AGE from Commander Squid liver fat when using hexane in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 7, except that hexane was taken as an organic solvent.

Received 0.109 kg of powder with an AGE content of 99.3% by weight.

The total yield of 54.0%.

The duration of the process was 11 hours.

EXAMPLE 18. Isolation of AGE from Commander Squid Liver Fat when hexane is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 8, except that hexane was taken as an organic solvent.

Received 0.119 kg of powder with an AGE content of 99.5% by weight.

The total yield of 59.2%.

The duration of the process was 11 hours 43 minutes.

EXAMPLE 19. Isolation of AGE from Commander Squid Liver Fat when hexane is used in the crystallization process.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 9, except that hexane was taken as an organic solvent.

Received 0.106 kg of powder with an AGE content of 99.2% by weight.

The total yield of 52.5%.

The duration of the process was 11 hours 54 minutes.

EXAMPLE 20. The allocation of AGE from liver fat of the commander squid when used in the process of crystallization of hexane.

To obtain AGE, fat was taken from squid liver in an amount of 1.0 kg. The content of AGE in marine fat is 20.0% by weight.

All operations in this example were carried out as in example 10, except that hexane was taken as an organic solvent.

Received 0.114 kg of powder with an AGE content of 99.5% by weight.

The total yield of 56.7%.

The duration of the process was 11 hours 7 minutes.

EXAMPLE 21. The allocation of AGE from the fat of the liver of king crab when used in the crystallization process of acetone.

To obtain AGE, fat was taken from the liver of king crab in the amount of 1.0 kg. The content of AGE in fat is 4.0% by weight.

All operations in this example were carried out as in example 1.

Received 0.020 kg of powder with an AGE content of 99.2% by weight.

The total yield of 50.6%.

The duration of the process was 11 hours 30 minutes.

The AGE preparations obtained in examples 1-21 contained in their composition by weight more than 90% of the sum of chemical, butyl and selachyl alcohols.

The total duration of the technological processes according to examples No. 1-21 was 11-12 hours.

As can be seen from the above examples, the inventive method allows to obtain the target product of good quality with high yield, while the method is not only technologically sophisticated, but also cost-effective, since it allows to reduce the duration of the process by five to six times (up to 11-12 hours) with minimal use of organic solvents during crystallization of AGE, and the remainder - free fatty acids - are also used in the future, which makes the processing of raw materials complex.

Filtered hexane or acetone solutions are regenerated by re-introducing the solvent into the process, and the bottoms remaining after evaporation and representing free fatty acids can be used in the production technology of dietary supplements Omegamarin (omega-3 polyunsaturated fatty acid concentrate, TU 9281-007 -02698223-10, Sanitary and epidemiological conclusion No. 77.99.11.003.T.000955.04.10 dated 04/07/2010; Patent of the Russian Federation No. 1581737 dated 05/21/1993 "Method for the production of ethyl ester concentrate eicosapentaenoic and docosahexaenoic acids", Kasyanov S.P. ., Latvian in NA, Glushchenko TV).

Claims (2)

1. A method of producing alkyl glycerol esters from sea fats, including hydrolysis of sea fat, acidification of the fat mixture, washing with water, first crystallization of the mixture from acetone or hexane, filtration, second crystallization of the precipitate from acetone or hexane, filtering and drying the target product, during the first and second crystallization, one of these solvents is used, characterized in that the ratio of the fat mixture to acetone or hexane having a temperature of from -15 ° C to -5 ° C, in the first and second crystallization stages, the composition It is 1: 1-5, kg / l. 2. The method according to claim 1, characterized in that the precipitates formed in the first and second crystallization stages, after filtration, are washed with acetone or hexane having a temperature from (0 ° C) to (+ 4 ° C).