RU2355403C1 - Method of obtaining biologically active product of young siberian stag antler breeding - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method includes raw material crushing, extraction with water solution of acidified ethanol at temperature 20-25°C with further extract freezing at temperature -15 - -18°C for 5 days and filtration of final product, as raw material blood fibrin of young Siberian antler deer is used, and process of extraction is carried out by means of 70% ethanol solution with ratio alcohol solution: raw material 10:1 for 5-6 days and daily extract heating to 50°C during 3-6 hours.

EFFECT: biologically active product with high activity, more rational use of young Siberian stag antler breeding.

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Description

The invention relates to the food and medical industry in the production of a biologically active product of antler reindeer husbandry, in particular from blood fibrin of antler deer.

A number of methods for producing biologically active products are known: pantohematogen (see Huntsman V.N., Deev N.G. "Antler reindeer husbandry" M .: Kolos, 1994, p. 52-53), the manufacture of which uses defibrinated blood of maral or spotted a deer; psycho-stimulating agent “Pantohematogen” (see RF patent No. 20088008, IPC АКК 35/14), the production of which also uses defibrinated blood. Fibrin in any production of medicines using defibrinated blood is a waste product and is disposed of.

A known method of obtaining a biologically active product (pantocrine), including the processing of antlers with hot steam, grinding, quadruple extraction with alcohol in a closed apparatus with steam heating, reflux condenser and mechanical stirrer. The first extraction is carried out four times (ratio of alcohol: raw materials 4: 1) in the amount of 96 ° ethyl alcohol, acidified with 1% acetic acid, for 6 hours. After cooling, the 2nd, 3rd, 4th extraction is carried out under the same conditions with 80% alcohol, but with an alcohol: feed ratio of 3: 1 at a temperature of 78 ° C. Alcohol is distilled off from the combined extract, the residue is used for the manufacture of tablets (see RF patent No. 195049, А61К 19/02).

However, a high concentration of ethyl alcohol used in this method and a temperature of 78 ° C contribute to the denaturation of most of the protein, including the protein part of hormones and enzymes, reducing the biological activity of the extract.

A known method of obtaining a biologically active extract (pantocrine) from antlers of red deer, red deer and sika deer (see FS-42-2323-85), including grinding antlers, three-time extraction (three times each) with 50% ethyl alcohol with periodic ( 5 minutes every hour) by stirring and subsequent filtration of the target product.

However, the technological parameters of this method do not provide high quality of the final product in the production of a biologically active product from fibrin obtained after the production of defibrinated blood.

A known method of obtaining a biologically active product of antler reindeer husbandry, including grinding raw materials, extraction with an aqueous solution of ethyl alcohol, periodically stirring the mixture and filtering the final product, where the penises and / or antler deer testicles are used as raw materials, and the extraction is carried out with an acidified 60% ethyl solution alcohol at a temperature of 20-25 ° C and a ratio of alcohol: raw materials of 5: 1 for 144-240 hours (6-10 days), while after the end of the process, the extract is subjected to freezing at a temperature (-15) - (-18) ° C for 5 days, followed by filtration of the final product. Stirring is carried out every 8 hours for 5 minutes (see patent application No. 2005123132, publication 01.27.2007 - prototype).

However, the method according to the prototype, when used in the production of a biologically active product from fibrin having a protein structure, and in the presence of a sedimentary part of the shaped elements, did not provide a high yield of biologically active substances during extraction.

It is necessary to develop a method for producing a biologically active product from fibrin obtained after the production of defibrinated blood, which provides a high yield of biologically active substances, which allows to expand the arsenal of agents with biological activity, and to rationally use the products of antler reindeer husbandry.

Studies to identify the presence of biologically active substances in fibrin after the production of defibrinated antler deer blood showed that the physicochemical and amino acid composition of fibrin is higher than in antlers (table 1 and 2).

Table 1 The content of the main physical and chemical indicators Indicators, % Fibrin Pant maral Water 14.97 14,99 Fat 10.83 1.26 Protein 70.75 43.65 Ash 3.45 40.10

table 2 Amino acid composition of samples Amino acid,% Fibrin Pant maral 1. Threonine 2.59 2.45 2. Tryptophan 0.58 0.41 3. Serine 1.25 0.94 4. Isoleucine 1.28 2.12 5. Glycine 1.34 1.00 6. Alanine 1,67 1.18 7. Valin 1.46 1.01 8. Methionine 1.10 1.09 9. Leucine 3.14 3,51 10. Glutamine 4.46 2.91 11. Proline 1.93 0.42 12. Phenylalanine 1,11 0.75 13. Lysine 3.87 3.94 14. Arginine 1.40 1.44 15. Methionine + Cystine 2.03 1.90 13. Total 29.21 25.07

As can be seen from table 1, the protein content in fibrin is 38.3% higher than in antlers, and according to table 2, the total amino acid composition is 14.2% higher.

To identify the optimal parameters of the fibrin extraction process, the influence of the following technological factors was investigated: extraction with 60, 70, 80% alcohol with a ratio of the components of the alcohol solution: raw materials of 2.5: 1; 5: 1; 10: 1 (hereinafter - dilution) and the duration of extraction of 3, 4, 5 and 6 days. The extraction process was carried out with temperature fluctuations during the entire period from 20 to 25 ° C (room temperature). The total values of amino acids (%) when extracting fibrin, crushed to the state of minced meat, are shown in table 3.

Table 3 Duration of extraction, days The total value of amino acids,% 2.5: 1 5: 1 10: 1 60% 70% 80% 60% 70% 80% 60% 70% 80% 3 6.67 8.70 7.68
(11.7) 8.56 10.13 8.84
(12.7) 13.33 15.67 13.60
(13.2) four 8.61 10.73 8.93
(16.8) 10.53 12,43 9.99
(19.6) 15.01 18.79 14.90
(20.7) 5 10.98 13.56 10.95
(19.2) 14.82 16.18 12.92
(20.1) 17.83 23.01 18.15
(21.1) 6 11.68 13.96 11.20
(19.8) 14.89 16.26 12.86
(20.9) 19.36 23.35 18.36
(21.4) Note: (...) the percentage reduction when extracted with 80% alcohol relative to the experience with 70% alcohol.

As can be seen from table 3, in all cases, the total amino acid value increased with increasing dilution (from 2.5: 1 to 10: 1), the duration of the process from 3 to 6 days, and with an increase in alcohol concentration from 60 to 70%. The use of 80% alcohol as an extractant had a negative effect on the yield of amino acids; moreover, the larger the dilution and the longer the extraction process, the higher the percentage reduction in the yield of amino acids. So, for example, with a dilution of 2.5: 1 and a process duration of 3 days, the percentage reduction in the yield of amino acids when extracted with 80% alcohol relative to the experiment with 70% alcohol was at the level of 11.7%, and with a duration of 6 days - 19 , 8%, with a dilution of 2.5: 1 - the percentage reduction is 11.7%, and with 10: 1 - 13.2%. According to studies, the optimal exposure of the fibrin extraction process to be considered is the use of a 70% ethanol solution as an extractant with a ratio of alcohol: feedstock ratio of 10: 1 and an extraction process duration of 5-6 days with amino acid yield values of 23.01-23.35% .

The results of subsequent studies on the effect on the output of amino acids of daily short-term (within 1.5; 3.0; 6.0 hours) heating of the extract to 50 ° C with more significant parameters of the fibrin extraction process determined in the first experiment (alcohol solution ratio: raw materials 5: 1 and 10: 1; alcohol concentrations of 60 and 70%; extraction times of 4; 5 and 6 days) are shown in table 4.

Table 4 Duration of extraction, days The total value of amino acids,% 5: 1 10: 1 60% 70% 60% 70% 1,5 hour 3.0 hour 6.0 hour 1,5 hour 3.0 hour 6.0 hour 1,5 hour 3.0 hour 6.0 hour 1,5 hour 3.0 hour 6.0 hour four 12.7 16,2 16.9 14.3 19.3 19.9 16.3 19.8 20.3 18.6 22.3 23.1 5 14.9 18,4 19.2 16.5 21.6 22.2 19.8 23.7 24.1 23.5 27.1 28.0 6 16.6 19.6 21.1 19.1 22.3 23,2 22.8 24.9 26.2 25.7 27.8 28.6

As can be seen from table 4, short-term heating of the extract to 50 ° C in all cases had a positive effect on the total yield of amino acids. Especially high growth dynamics of this indicator when heated from 1.5 to 3 hours, so with the duration of the extraction process for 5 days with daily heating of the extract to 50 ° C for 1.5 hours (60% alcohol) the amount of amino acids It was at the level of 14.9%, and when heated for 3 hours - 18.4% (an increase of 23%), while 6 hours of heating provided an increase of this indicator relative to 3-hour heating by only 4.7 % The content of vitamins in 60% extracts increased with increasing extraction time and the duration of heating the solution by 7.8-12.4%, however, the concentration of vitamins was 12.9-17.3% lower than that of similar 70% extracts .

According to studies, the optimal exposure of the fibrin extraction process using short-term heating should be considered as the process using 70% ethanol as an extractant with a ratio of alcohol: raw materials of 10: 1 and a process duration of 5-6 days and with a single daily heating of the extract to 50 ° C for 3-6 hours, which provides an amino acid yield of 27.12 to 28.65%, while the optimal exposure of the extraction process without heating (experiment, table 3), with the same process parameters, is higher - 23.01-23.35%, which is on average 19.2% less than when heated, and at the maximum rates of two experiments (23.35 and 28.65%), this excess is 22.7%.

The activity of the drugs was determined by the biological method according to their ability to lower blood pressure in rabbits, as well as by the assessment of adaptogenic (duration of stay of the animal at + 2-4 ° C) and tonic effect (duration of swimming of the animal until muscle fatigue).

The results of checking the effectiveness of the proposed method are shown in table 5.

Table 5 Raw materials and extraction method Extraction speed, hours The biological activity of the extracts according to the standard FS 42-2323-85 Adaptogenic action, min Tonic effect, min 1x introduction 5x introduction 1x introduction 5x introduction penises and / or testes - prototype 144-240 25-32% 153 174 63 78 blood clots (fibrin) 120-144 35% 189 203 105 167

According to table 5, the extract from blood clots (fibrin) obtained after defibrination of blood exceeds the extract from penises and / or testes (prototype) in hypotensive effect by 8.6-28.6%, adaptogenic in 14.2-19.1% and tonic - 40.0-46.7%.

Example. The crushed (for example, on the cutter) fresh (collected on the day of the maral slaughter) blood clots (fibrin) obtained after defibrination and blood filtration were mixed with a 70% aqueous solution of ethyl alcohol (acidified with 1% acetic acid to pH 4, 7) in the ratio of a solution of alcohol: raw materials 10: 1 and was extracted for 5 days at room temperature (+ 23 ° C). Moreover, every 24 hours, the extractant was heated to 50 ° C for 3 hours. After the required extraction time, the extract was placed in a refrigerator with a temperature of -18 ° C for 5 days for freezing, followed by filtration of the final product.

Thus, the use of the claimed method of producing a biologically active product from blood clots (fibrin) remaining after defibrinization of antler deer blood allows one to obtain a drug that exceeds the prototype by hypotensive, adaptogenic and tonic effect, to simplify the extraction process (one-time mixing of alcohol with raw materials instead of 3 multiple of the prototype with daily heating of the working solution instead of 24-fold mixing of the prototype), expand the arsenal of products with biological active activity, and to use antler reindeer products more fully and rationally.

Claims (1)

A method of obtaining a biologically active product of antler reindeer husbandry, including grinding of raw materials, extraction with an aqueous solution of acidified ethyl alcohol at a temperature of 20-25 ° C, followed by freezing of the extract at a temperature of (-15) - (- 18) ° C for 5 days and filtering the final product , characterized in that the blood fibrin of antler deer is used as a raw material, while the extraction process is carried out with a 70% solution of ethyl alcohol with a ratio of alcohol solution: feedstock 10: 1 for 5-6 days and daily heating of extra act to 50 ° C for 3-6 hours