The method of obtaining funds with antitoxic and anti-tumor activity

 

(57) Abstract:

The invention relates to the field of medicine and is concerned with the obtaining medicines from animal waste materials. The method includes crushed spleen of cattle, the influence extracting agent, separating the extract and subsequent removal of the extractant. As extractant used an aqueous solution of ethyl alcohol. The effect of extractant complement the simultaneous effect of low frequency ultrasound. The separation of the extractant carried out by centrifugation. Effect: method provides increased product yield while reducing the time of its receipt. 6 C.p. f-crystals, 3 tables.

The invention relates to the field of medicine and is concerned with the obtaining medicines from animal waste materials.

Known methods for producing biologically active substances and drugs of animal waste material using organic solvents [1,2].

The closest in technical essence is a method of obtaining medicines (splenin) from the spleen of cattle with the use of dichloroethane by infusion [3-5].

In the reactor under vacuum (6910 kPa) served with minced meat and solvent (dichloroethane) in a weight ratio of 1:2 (one part meat and two parts of dichloroethane). The extraction is carried out at a temperature of 20oWith by infusion over a period of 5-6 days, using initial mixing. After extraction, the mixture is settled and separated into two layers. Then separate declaratively extract from the pulp by filtration through filter fungus under vacuum. From the extract to remove the dichloromethane by distillation under vacuum 6910 kPa at a temperature of 45oC. the resulting aqueous extract is treated with 70% ethanol and petroleum ether. The ether fraction is separated and the aqueous-alcoholic fraction remove the alcohol by distillation under vacuum. Water extract was added to 0.9% (mass) NaCI brand OFS (especially pure). The contents shaken until complete dissolution NaCI, stand in the refrigerator at a temperature of 61oC, 141 hours. After vysalivaniya the contents filtered. The resulting aqueous extract is diluted with a physiological solution with subsequent preservation of ethyl alcohol in the range 9-11 volume percent. Of 552 kg of meat of bovine spleen receive 3,60,1 kg BW of an aqueous extract of splenin, which in terms of Isho substances, having biological activity;

- the long duration of the extraction process;

- the use of environmentally harmful solvents (dichloromethane, petroleum ether).

The objective of the invention is to provide a method of obtaining funds with antioxidant and radioprotective activity, with increased output and reduced duration.

In accordance with the task proposed method of obtaining funds with antioxidant and radioprotective activity includes grinding the spleen, the influence extracting agent, separating the extract and subsequent removal of the extractant and differs from the prototype in that as the extracting agent is used an aqueous solution of ethyl alcohol, simultaneously with the specified exposure extracting agent provide exposure to low-frequency ultrasound, and separating the extractant carried out by centrifugation.

In the preferred embodiment of the method of removing the extractant from the product centrifugation in terms of its alcohol component is carried out by distillation of the alcohol under vacuum at a temperature not exceeding 45oC. thus p is m the embodiment of the method of removing the extractant from the product of the centrifugation is carried out by distillation of the alcohol under vacuum at a temperature not exceeding 45oC and subsequent lyophilization of the obtained aqueous extract. At the same time as the final product get dry extract in powder form with a longer shelf life.

It is advisable that an aqueous solution of ethyl alcohol contained 65 to 75 weight percent ethyl alcohol. Within these concentrations provided the greatest yield of the final product at the lowest duration of the extraction process. When alcohol concentration is less than 65% of the extraction process is unacceptable longer, and when alcohol concentration greater than 75% decreases the yield of biologically active substances.

It is advisable to choose the weight ratio of the spleen and of the extractant from the interval 1/6 - 1/3, which provides the greatest yield of the final product and the lowest duration of the extraction process under other identical conditions. The excess of the specified weight ratio 1/3 reduces the specific yield of the final product. When the weight ratio of spleen to the extractant smaller 1/6 increases the amount and duration of the removal process of the extractive agent, this reduces expressnet method in General. Upon receipt in the quality of the final product aqueous extract additional umanage low-frequency ultrasound to choose between 18-70 kHz. In the specified frequency range under other equal conditions is the most comprehensive and quick extraction. Outside of a selected range of frequencies, the impact of ultrasound decreases, which leads to an unacceptable reduction in product yield or increase the duration of extraction.

It is preferable to choose the intensity of the specified low-frequency ultrasound in the interval of 0.1 to 2.3 W/cm2within which the extraction efficiency is maximum. When the intensity is less than 0.1 W/cm2greatly reduces the efficiency of extraction. When the intensity of the ultrasound more than 2.3 W/cm2the extraction efficiency slowly decreases with increasing energy.

Next, the proposed method of obtaining funds from the antioxidant and radioprotective activity from bovine spleen is described on the example of the preferred embodiment.

Fresh or frozen spleen clear of a lipid film and crushed, for example with the grinder. To the resulting milled mass add five weight parts of 70% aqueous ethanol solution, the resulting mixture is placed in ultrawellness.com radiation with a frequency of 18 kHz and an intensity of 2 W/cm2. After 45 minutes the contents of the ultrasonic bath is placed in a centrifuge where they separate the extract from the pulp of raw materials. Next product centrifugation to remove the extractant in the alcohol component by distillation under vacuum 6910 kPa at a temperature of 45oC.

Next, an aqueous solution of the extract and the foundations of the prototype process NaCI brand OFS and filter that helps to remove proteins. In this form the aqueous solution of the obtained extract can be used to obtain the ampoule liquid dosage forms. To improve ease of use, storage and transportation, an aqueous solution of the extract is subjected to freeze drying and receive a dry white powder with a yield of biologically active substances of about 5.4% by weight of the feedstock.

Other examples of the proposed method with different modes and conditions of implementation are summarized in table. 1.

Examples carried out with the same duration of extraction (45 minutes) under the action of ultrasound. The exception is the example 7, in which the period of extraction has been extended to 70 minutes. As seen from the above examples within the optimal range of the weight of soothes the different frequencies 18-70 kHz and an intensity of 0.1 to 2.3 W/cm2, exposure to ultrasound, the proposed method provides a yield of dry product, exceeding 5% by weight of the feedstock. Outside these intervals, the product yield relative to the source of raw materials is less than 5%, although in this case exceeds the output by way of a prototype in order and more.

Comparative trials of pharmacological activity and acquisition parameters preparations obtained from the spleen of cattle in the method prototype (splenin) and proposed method (esplin), the results of which are compared with the control observations are summarized in table. 2 and 3.

Received the tool according to the proposed technical solution antioxidant and radioprotective activity is not inferior, and in some settings antitumor activity and normalization of nitrogen metabolism exceeds splenin (see tab. 2).

The use of the claimed invention will:

to increase the yield of the target product;

to increase saving;

- to improve the environmental situation in obtaining the product.

At the same time, the proposed method allows you to save, and on some parameters to improve the biological is Sidorova, C. L. Konstantinov, O. C. Gull, N.F. Towarowa. Auth. St. N 1228776, published. 30.04.86. Bull. N 6.

2. Andras Balagi, Mihai Saigo, Lajos Kisfaludi, Tibor Klupp, Miklos Barabas. Patent Hungary 162447, CL V 01 N 13/00, 1967.

3. Industrial regulations on Splenin (substance) N 64-04-34-36, 1984.

4. Pharmacopeia 42-1255-90. Splenin, the USSR Ministry of health, Formamide, 10.07.91.

5. State Pharmacopoeia of the USSR, the USSR Ministry of health, 11th Edition, M, Medicine, 1990.

1. The method of obtaining funds with antitoxic and anti-tumor activity of the spleen of cattle, including grinding the spleen, the influence extracting agent, separating the extract and subsequent removal of the extractant, characterized in that as the extracting agent is used an aqueous solution of ethyl alcohol, this energy extracting agent is complemented by simultaneous exposure to low-frequency ultrasound, Department of the extract is carried out by centrifugation.

2. The method according to p. 1, characterized in that it is removing the extractant from the product of the centrifugation is carried out by distillation of the alcohol under vacuum at a temperature not exceeding 45oC.

3. The method according to p. 1, otli under vacuum at a temperature not exceeding 45oC and subsequent lyophilization of an aqueous extract.

4. The method according to PP.1 to 3, characterized in that it is an aqueous solution of ethyl alcohol contains 65 to 75 wt.% ethyl alcohol.

5. The method according to PP.1 to 4, characterized in that in it the weight ratio of spleen and extractant selected from the interval of 1/6 - 1/3.

6. The method according to PP.1 to 5, characterized in that it specified frequency ultrasound is chosen from the interval 18 - 70 kHz.

7. The method according to PP.1 - 6, characterized in that it specified the impact of ultrasound exercise intensity of 0.1 to 2.3 W/cm2.

 

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3 ex, 2 tbl

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1 ex, 1 tbl

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1 tbl

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12 cl, 4 ex, 3 tbl

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