RU2353375C2 - Method of obtaining biologically active product with otensive activity - Google Patents

Abstract

FIELD: animal husbandry.

SUBSTANCE: present invention relates to processing deer products, particularly to methods of obtaining a biologically active pantocrine type product from fresh genital organs of deer. The method involves grinding the raw material, extraction using an aqueous solution of ethanol, periodic stirring of the mixture and filtration of the end product. The raw products are penises and/or testicles of deer. Extraction is done through acidification with a 60% solution of ethanol at 20-25°C in the ratio alcohol: raw material 5:1 for 144-240 hours. The mixture is stirred every 8 hours for 5 minutes. After extraction, the extract is frozen at temperature -15…-20°C for 5 days with subsequent filtration of the end product.

EFFECT: invention widens the raw material base, simplifies the process in conditions which provide for high biological activity of the end product.

3 tbl, 1 ex

Description

The invention relates to the processing of reindeer herding products, in particular to methods for producing a biologically active product such as pantocrine from fresh genital organs of antler deer, namely from penises and testes of antler deer.

A known method of obtaining a biologically active product from antlers and / or blood, and / or the internal organs of antler deer, including grinding the raw component, drying the minced meat under vacuum and grinding to a powder state. Drying, combined with grinding, is carried out at a temperature not exceeding 60 ° C and a vacuum of 10-60 mm Hg. However, the powder obtained by this method can be used only for bathing (see RF patent 2130777, AK 35/32).

A known method of obtaining a biologically active product from antlers of maral (pantocrine) by hydrolysis of antler flour (ground antlers) 6 N. hydrochloric acid at a temperature of 160 ° C for 24 hours. However, this method allows only 0.8% of the free amino acids concentrated in the antler to be extracted, which mainly determine the biological activity of the product (see the book Pantokrin, Gorno-Altaysk, 1969, pp. 30-32). In addition, the use of hydrochloric acid (HCl) in this method further determines its neutralization and laborious removal of the resulting salt from the hydrolyzate.

A known method for producing pantocrine, including the treatment 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 (alcohol: raw material ratio 4: 1) in an 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 deer, red deer and sika deer (in accordance with FS-42-2323-85), including grinding antlers, three times extraction (three times each) for 50% ethyl alcohol with periodic (5 minutes every hour) by stirring and subsequent filtration of the target product (see Industrial regulation No. 35 for pantocrine production, Minmedbioprom, Moscow endocrine plant, 1986) prototype.

However, studies have shown that when using antler deer (penises and / or testes) as a raw material of the genitals of the deer (penises and / or testes), the use of a 50% aqueous alcohol solution does not ensure complete extraction, and a 3-fold exposure to a new portion of alcohol also worsens the biological activity of the final product and complicates the process.

It is necessary to develop a method for producing a biologically active product, which allows to expand the raw material base, to simplify the process under a regime that ensures high biological activity of the final product.

The specified technical result is achieved by the fact that in the method of producing a biologically active product of antler reindeer husbandry, including grinding raw materials and extraction with ethyl alcohol, penis and / or antler deer testes are used as raw material, and extraction is carried out acidified to pH 4.7 with a 60% solution ethyl alcohol at room temperature 20-25 ° C and the ratio of alcohol: raw materials 5: 1 for 144-240 hours. After the extraction process is completed, the extract is subjected to freezing at a temperature of -15 to 18 ° C for 5 days, followed by filtration of the final product.

To identify the optimal process parameters, the influence of factors was studied: the concentration of ethyl alcohol (50, 60, 70%), the ratio of the components of alcohol: raw materials - 3: 1; 4: 1; 5: 1, extraction time - from 48 to 240 hours when carrying out the extraction process at room temperature 20-25 ° C.

Table 1 shows the total amino acid values (g / kg) (iso-leucine, threonine, serine, glycine, alanine, valine, methionine, leucine, glutamine, proline, phenylalanine, lysine, arginine) when extracted with 60- and 70% ethyl alcohol crushed to a state of minced penis.

Table 1 The duration of extraction, hours Alcohol: Raw Material Ratio 3: 1 4: 1 5: 1 60% 70% 60% 70% 60% 70% 48 15.08 16.00 15,56 16.63 16.30 16.71 72 15.61 16.67 16.40 16.83 16.81 16.16 144 15.79 16.90 16.82 17.08 17.38 15.40 240 16.68 17.06 17.21 17.26 18.36 15.30

As can be seen from table 1, the amino acid content during extraction with 60% alcohol increased with increasing duration of extraction and the amount of alcohol in the extract. Due to the fact that the rate of increase (percentage of increase from exposure to exposure) of the amount of amino acids decreased significantly with a change in exposure, further expansion of the field of study of process parameters is impractical. So, with a ratio of 3: 1 and 48 hours of extraction, the total amino acid value was 15.08 g / kg, with a ratio of 4: 1 - 15.56 g / kg (an increase of 3.2%), while when switching from 4: 1 ratio to 5: 1 ratio the increase in the amount of amino acids was only 1%. A similar situation was observed when studying the optimal duration of extraction. The result of extraction with 70% alcohol at an alcohol: feed ratio of 3: 1 and 4: 1 was slightly higher than 60%, and reached its maximum value of 17.26 g / kg at a ratio of 4: 1. However, a further increase in the amount of alcohol (5: 1) and the duration of extraction from 72 to 240 hours significantly reduced (probably due to the process of protein denaturation) the total amount of amino acids. Therefore, the optimal parameters of the mode of extraction of penises are 60% alcohol as an extractant with an alcohol: raw material ratio of 5: 1 and extraction time (duration) of 144-240 hours. The total value of amino acids in the extract with these parameters is 17.38-18.36 g / kg.

Table 2 shows the total values of amino acids when extracted with 60- and 70% ethyl alcohol, crushed penises with testes minced to the state of minced meat with the same extraction duration parameters and component ratio.

Table 2. The duration of extraction, hours Alcohol: Raw Material Ratio 3: 1 4: 1 5: 1 60% 70% 60% 70% 60% 70% 48 13.52 13.66 14.80 14.10 15.38 14.18 72 13.72 13.84 15.92 15.81 15.97 15.96 144 14.45 14.68 16,44 16.27 17.56 15.61 240 14.83 15,55 17.25 17.73 19.16 15.63

As can be seen from table 2, during the extraction of penises with testes, the course of the extraction process was similar to the process of extraction of penises. The optimal total amino acid indices were slightly higher (17.56-19.16 g / kg) with the same process parameters: extractant - 60% alcohol, extraction duration 144-240 hours with an alcohol: feed ratio of 5: 1. The experiments were carried out in both cases at room temperature of 20-25 ° C. The results of extraction of penises and / or testes with 50% ethanol at all three ratios studied were significantly lower. The content of vitamins in 60% extracts from penises with a ratio of alcohol: raw materials of 3: 1; 4: 1; 5: 1 exceeded this indicator in 70% extracts by 16.05; 18.30; 20.90%, respectively, in extracts from penises with testes - by 4.90; 4.20; 3.60%. Due to the fact that during the first mixing of minced meat from penises and / or testes, a long non-settling suspension formed, stirring was performed 3 times a day for 5 minutes.

The results of the study on the use of testes as a raw material in the production of a biologically active product are not given in the description due to the fact that the data on the use of penises with testes as raw materials were higher than when using penises, which means that some testes are obviously more effective as raw materials for effectiveness, which is confirmed in the description (table 3).

Acidification of the extracts to a pH of 4.7 ensured the subsequent precipitation of long-cyclic protein molecules into a precipitate upon freezing. The freezing temperature from -15 to -20 ° C is due to the fact that at a temperature above -15 ° C the process of protein deposition is longer, and below -20 ° C crystallization of bound water occurs, which further reduces the activity of the drug.

The results of checking the effectiveness of the proposed method are shown in table 3.

Table 3 Raw materials and extraction method Extraction speed, hours The biological activity of the extracts according to the standard FS 42-2323-85 Antlers of maral, prototype method 216 25% standard Penis, the proposed method 144-240 25-27% Penises with testes, the proposed method 144-240 26-29% Testes, the proposed method 144-240 29-32% Penises with testes according to the prototype method 216 23%

The activity of the drugs was determined by the biological method according to their ability to lower blood pressure in rabbits.

Example. Shredded (for example, on a cutter) fresh (collected on the day of the maral slaughter) penises with testes were mixed with a 60% aqueous solution of ethyl alcohol (acidified with 1% acetic acid to pH 4.7) in the ratio of alcohol: raw materials 5: 1 and was extracted for 168 hours at room temperature + 23 ° C. Moreover, every 8 hours, the mixture was stirred for 5 minutes. After the required extraction time, the extract was placed in a refrigerator with a temperature of -18 ° C for 3 days for freezing, followed by filtration of the final product.

Thus, the use of the claimed method of obtaining a biologically active product from penises and / or testes of antler deer allows you to get a drug at a level higher in hypotensive effect with pantocrine, to simplify the extraction process (one-time mixing of alcohol with raw materials instead of 3 times the prototype at 3- mixing the mixture multiple times per day instead of the 24-fold prototype), expand the arsenal of products with biological activity and use antler products more fully and rationally herding.

Claims (1)

A method of obtaining a biologically active antler deer-breeding product having a hypotensive effect, including grinding the raw material, extracting with an aqueous solution of ethyl alcohol, periodically mixing the mixture and filtering the final product, characterized in that the raw materials are used penis and / or antler deer testicles, and the extraction is acidified 60% solution of ethyl alcohol at a temperature of 20-25 ° C and the ratio of alcohol: raw materials 5: 1 for 144-240 hours, mix the mixture every 8 hours for 5 minutes, while after about the end of the extraction process, the extract is subjected to freezing at a temperature of -15 ... -20 ° C for 5 days, followed by filtration of the final product.