RU2311191C2 - Lucerne extract - Google Patents

Abstract

FIELD: pharmaceutical industry.

SUBSTANCE: lucerne extract possessing immunomodulator properties is obtained through extraction of crude material with aqueous solution containing following trace minerals, mg/kg : iron 320.0, zinc 180.0, copper 45.0, manganese 25.0, cobalt 6.0, chromium 4.0, selenium 3.0, molybdenum 5.0, iodine 2.0, and fluorine 1.0. Extraction is carried out for 55-65 min at 30-40°C. Mixture is then heated to 122-142°C at pressure (2-3)·10⁵ Pa, held for 2 h, cooled, purified, and evaporated to thickness at 95-95°C.

EFFECT: increased biological activity of product.

2 cl, 2 tbl, 3 ex

Description

The invention relates to medicine, namely to the production of medicines and biologically active food supplements from plant materials with therapeutic properties, and can find application in the pharmaceutical, cosmetic and food industries, as well as in veterinary medicine.

Known drug (1), including alfalfa hay extract obtained by processing alfalfa hay with an extractant containing a mixture of trace elements in the following ratio of plant mass, mg / kg:

Mo 10.0 Pb 20,0 With 10.5 V 1,0 Cr 0.5 Zn 200,0 Fe 300,0 Sn 40,0

The extract is used to treat postoperative animal wounds.

The closest technical solution is the biologically active drug Dolutsar based on alfalfa extract (2) obtained by treating alfalfa hay with an extractant containing a mixture of trace elements salts in the following ratio of plant materials, mg / kg:

Mn 22.0 Wa 13.0 Sn 0.6 With 4.0 Cr 1,0 V 1.8 Zn 700,0 Fe 1030.0 Se 1,0 Cu 1,0

Dolucar extract has increased biological activity and immunomodulating properties. The disadvantage of this extract is the presence in the extractant of microelements toxic to humans (barium, vanadium and tin) and the absence of some vital (essential) microelements, which limits the possibility of using the extract for humans (3).

Barium is a microelement toxic to humans; entering the human body, it is deposited in the liver, brain, endocrine glands, and bones, turning into insoluble compounds. Acute intoxication with barium and its salts is characterized by profuse salivation, burning in the mouth and esophagus, abdominal pain, nausea, vomiting, diarrhea, high blood pressure, convulsions (possible paralysis), cyanosis, cold sweat, gait and speech disorders, shortness of breath, visual impairment. With increasing intoxication, death can occur within a day.

Vanadium also refers to micronutrients toxic to humans, entering the body, it is concentrated in the bones, kidneys, liver, spleen, intestines, stomach, testes, lungs and in the brain. Intoxication with vanadium and its salts is characterized by severe runny nose, sneezing, lacrimation, dry throat, chest pains, general weakness and headache. With increasing intoxication, bronchospasm, bronchitis, bronchopneumonia, cough, hemoptysis and bleeding develop, as well as allergic reactions in the form of bronchial asthma and eczema.

Tin refers to trace elements potentially toxic to humans. With body intoxication, this element is characterized by persistent headaches, vomiting, abdominal pain, visual and mental disorders, paresis.

Given the toxic properties and the absence of established signs of deficiency of barium, vanadium and tin in the human body, according to SanPiN 2.3.2.1078, these microelements are not included in the list of mineral substances (macro- and micronutrients) that are allowed for use in the manufacture of biologically active food additives, therefore enter they are additionally unacceptable in the body (4).

A known method of producing an extract from alfalfa (2), including processing plant materials with an extractant containing an aqueous solution of salts at a temperature of 60-80 ° C for 20 minutes, holding the extracting mixture at a temperature of 150 ° C and pressure (5.4-5.9 ) × 10 ⁵ Pa for 90 minutes with further separation, purification and drying of the extract at a temperature of 200-300 ° C.

To implement the method, the following equipment is used: a mixer equipped with a pump for pulp circulation; a reactor equipped with an integrated receiving tank for separating the obtained extract; centrifuge; evaporator; spray dryer and container for collecting the finished product. In addition, there is a container for paracondensate, a microelement dispenser, pipelines and a steam cooler.

The disadvantage of this method is the use of bulky and complex equipment, the design of which does not allow to remove parts in contact with raw materials, materials and intermediates for washing, processing with disinfectants and sterilization. This equipment cannot be used for the preparation of medicines and dietary supplements, as without such treatment, the obtained extract is not safe for human health. It is also impossible to use steam condensate, as in the manufacture of medicines and biologically active food additives, it is allowed to use only purified water or water for injection (5, 6). In addition, the dry extract is hygroscopic, and it is difficult to standardize for the subsequent preparation of dosage forms and dietary supplements.

The objective of the invention is to obtain a liquid extract from alfalfa with high biological activity and immunomodulating properties, non-toxic and safe for therapeutic and prophylactic use using industrial equipment for the manufacture of medicines and biologically active additives.

The essence of the invention is to obtain an extract from alfalfa using an extractant without condensate and toxic microelements and introducing water and vitally important (essential) microelements of molybdenum, iodine and fluorine into it, in the following ratio of raw materials, mg / kg:

Fe 320,0 Zn 180.0 Cu 45.0 Mn 25.0 Co 6.0 Mo 5,0 Cr 4.0 Se 3.0 I 2.0 F 1,0

Alfalfa grass or hay is used as raw material.

The essence of the invention also lies in the fact that in the method for producing the extract, the processing of raw materials is carried out with an aqueous solution of salts of trace elements: Fe, Zn, Cu, Mn, Co, Mo, Cr, Se, I, F, at a temperature of 30-40 ° C for 55 -65 minutes, and then the mixture is heated to a temperature of 122-142 ° C at a pressure of (2-3) × 10 ⁵ Pa, incubated for 2 hours, cooled, purified and evaporated at a temperature of 85-95 ° C to the desired concentration.

Unlike the prototype, the extractant does not contain condensate and toxic microelements, but purified water is used or water for injection with vital microelements dissolved in it. In addition, when preparing the extract, the processing of the raw material with the extractant (maceration) is carried out at a temperature of 30-40 ° C for 55-65 minutes, the mixture is heated to a temperature of 122-142 ° C and at a pressure of (2-3) × 10 ⁵ Pa, withstand for 2 hours, and evaporation is carried out at a temperature of 85-90 ° C to the required concentration.

The absence of condensate and toxic trace elements Ba, V and Sn in the extractant ensures the safety of the extract for humans.

The use of water as an extractant increases the degree of swelling of some biopolymers and accelerates the reaction of increasing molecular weight due to its specific properties (high dielectric constant, solvation ability, the presence of intermolecular hydrogen bonds, providing a relay mechanism for proton transfer, etc.). The introduction of inorganic salts increases the solvent capacity of the extractant, accelerates the polymerization reactions, and significantly increases the specific viscosity of the extract (7).

The introduction of the vital trace elements molybdenum, iodine and fluorine into the extractant is due to numerous diseases of people associated with a deficiency of these trace elements (3).

Molybdenum is an essential mineral for humans, which performs its biological function as an integral part of the three molybdenum-containing enzymes aldehyde oxidase, xanthine oxidase and sulfite oxidase. Lack of molybdenum in the body disrupts the synthesis of each of these enzymes, which leads to urolityliasis, pronounced brain abnormalities, mental retardation and ectonia of the lens. Molybdenum deficiency is one of the high risk factors for people with esophageal tumors.

Iodine is a true biomicroelement that fulfills its biological function as an integral part of thyroid hormones (thyroxine and triiodothyronine), which perform vital functions, are responsible for metabolism throughout the human body, controlling the consumption of proteins, fats and carbohydrates. These hormones regulate the activity of the brain, nervous system, genital and mammary glands, and the growth and development of children. Lack of iodine in adults leads to endemic goiter, and with prolonged iodine deficiency in children leads to criticism, underdevelopment of the brain and skeletal system.

The biological effect of fluoride ion is based on its ability to effectively replace hydroxyl ion in apatite of bone tissue, in non-mineralized tissues and in the active center of enzymes. A lack of fluoride is one of the most widespread pathological processes of modern mankind, leading to a delay in teething and specific damage to teeth by caries in children; In adults, it also causes dental caries, and in old age - fluoride-dependent osteoporosis of the skeleton and a tendency to fractures, especially in women.

The number of micronutrients introduced into the extract and their ratio is established taking into account their amount in alfalfa grass and the daily physiological need for these micronutrients in humans.

Lowering the temperature of processing raw materials with an extractant to 30-40 ° C allows you to save the biological activity of heat-sensitive biopolymers extracted from raw materials.

Heat treatment of the extract at a temperature of 122-142 ° C and a pressure of (2-3) × 10 ⁵ Pa for 2 hours ensures reliable sterilization, while in addition to killing microorganisms, adiabatic compression of the mixture and associated ballast high molecular weight compounds (mucus, pectins, starch, etc.) become insoluble, which are then removed by filtration.

The purified extract is evaporated at a temperature of 85-95 ° C to the required concentration, while the condensation polymerization of the extract takes place and the introduced trace elements bind through active groups to organic compounds extracted from alfalfa grass, forming strong macromolecules. Evaporation at a temperature of 85-95 ° C eliminates the possibility of spontaneous isolation (precipitation) of the polymer from the extract and makes it easy to standardize the extract, to ensure its antibacterial stability during prolonged storage.

The proposed method ensures the safety of the obtained extract, because allows you to use easily sterilized industrial equipment: a reactor (equipped with a steam jacket, cooling system and a sealed cap), a filter and an evaporator, approved for use in the manufacture of medicines and biologically active food additives.

A method of obtaining an extract of alfalfa is as follows.

The dried and crushed alfalfa grass, previously passed the input control, is weighed, loaded into the reactor and the extractant containing the trace elements salts diluted in water is fed: Fe, Zn, Cu, Mn, Co, Mo, Cr, Se, I and F. The reactor is hermetically closed , the mixture is heated to a temperature of 30-40 ° C and extraction is carried out by fractional maceration for 55-65 minutes. After that, the mixture in the reactor is heated to a temperature of 122-142 ° C and maintained at a pressure of (2-3) × 10 ⁵ Pa for 2 hours. Then the mixture is cooled to a temperature of 30-40 ° C, and the pressure in the reactor is reduced to atmospheric. From the reactor, the extract is filtered into the evaporator, cleaning it of the associated ballast substances and mechanical impurities. In the evaporator, the purified extract is evaporated at a temperature of 85-95 ° C to the desired concentration. After evaporation, the extract is cooled, standardized, dispensed in sterile conditions into vials, sealed and transferred to a control.

The extract obtained in this way in bottles is sterile, harmless and is a dark brown liquid, stable for at least 3 years. The study of acute and chronic toxicity in laboratory animals with various routes of administration showed that the extract obtained is practically non-toxic and does not have a locally irritating, skin-sensitizing, allergenic, embryotoxic and teratogenic effect. It is used for the preparation of various dosage forms and dietary supplements.

The proposed method is implemented and represented by the following examples.

Example 1

In a 100-liter reactor equipped with a steam jacket, a cooling system and a hermetically sealed lid, 8 kg of dried and crushed alfalfa grass are loaded and 52 l of extractant containing trace elements dissolved in water for injection: Fe, Zn, Cu, Mn, Co, Mo, Cr, Se, I and F. The reactor is sealed with a lid, the mixture is heated to a temperature of 30 ° C and extracted at this temperature for 65 minutes. After that, the mixture in the reactor is heated to a temperature of 142 ° C and maintained at a pressure of 3 × 10 ⁵ PA for 2 hours. Then the mixture in the reactor is cooled to a temperature of 30 ° C, after relieving the pressure, the lid is opened and the extract from the reactor is filtered into an evaporator. In an evaporator with a capacity of 60 l, the purified extract is evaporated at a temperature of 85 ° C to a concentration of 50%. After evaporation, the extract is cooled, standardized, dispensed in sterile conditions into vials, sealed and transferred to a control. The extract obtained is used for the preparation of biologically active food additives.

Example 2

In a 100-liter reactor equipped with a steam jacket, a cooling system and a hermetically sealed lid, 8 kg of dried and crushed alfalfa grass are loaded and 52 l of extractant containing trace elements dissolved in water for injection: Fe, Zn, Cu, Mn, Co, Mo, Cr, Se, I and F. The reactor is sealed with a lid, the mixture is heated to a temperature of 35 ° C and extracted at this temperature for 60 minutes. After that, the mixture in the reactor is heated to a temperature of 132 ° C and maintained at a pressure of 2.5 × 10 ⁵ Pa for 2 hours. Then the mixture in the reactor is cooled to a temperature of 35 ° C, after relieving the pressure, the lid is opened and the extract from the reactor is filtered into an evaporator. In an evaporator with a capacity of 60 l, the purified extract is evaporated at a temperature of 90 ° C to a concentration of 50%. After evaporation, the extract is cooled, standardized, dispensed in sterile conditions into vials, sealed and transferred to a control. The resulting extract is used for the preparation of dosage forms.

Example 3

In a 100-liter reactor equipped with a steam jacket, a cooling system and a hermetically sealed lid, 8 kg of dried and crushed alfalfa grass are loaded and 52 l of extractant containing trace elements dissolved in purified water are added: Fe, Zn, Cu, Mn, Co, Mo , Cr, Se, I and F. The reactor is sealed with a lid, the mixture is heated to a temperature of 40 ° C and extracted at this temperature for 55 minutes. After that, the mixture in the reactor is heated to a temperature of 120 ° C and maintained at a pressure of 2 × 10 ⁵ Pa for 2 hours. Then the mixture in the reactor is cooled to a temperature of 40 ° C, after relieving the pressure, the lid is opened and the extract from the reactor is filtered into an evaporator. In an evaporator with a capacity of 60 l, the purified extract is evaporated at a temperature of 95 ° C to a concentration of 50%. After evaporation, the extract is cooled, standardized, dispensed in sterile conditions into vials, sealed and transferred to a control. The resulting extract is used for the preparation of dosage forms.

Physico-chemical characteristics of alfalfa planting alfalfa "Luceron" obtained by the proposed method are shown in table 1.

Comparative tests of extracts "Dolucar" (prototype) and "Luceron" obtained by the proposed method (table 2) showed that Luceron in terms of biologically active substances and biological activity exceeds Dolucar.

Table 1
Physico-chemical characteristics of alfalfa extract "Luceron" Name of indicator Method implementation examples Example 1 Example 2 Example 3 pH 5.3 5.1 5.1 Density, g / cm ³ 1.22 1.20 1.24 Viscosity (pz) 0.65 0.64 0.66 Total ash,% 17.8 20.7 22.7 The content of heavy metals,% no more 0.001 0.001 0.001 The content of vitamin B ₂ , mg / ml 24.2 39.25 29.41 The content of vitamin B ₁₂ , mg / ml 30.5 29.8 23.8 The content of vitamin C, mg / ml 5.22 4.46 3.57 The content of vitamin E, mg / ml 11.7 11.8 12 The content of uronic acids, mg / ml 10.8 13.5 15.6 Flavonoid content, mg / ml: - according to luteolin 9.9 9.5 10,4 - by routine 13.5 12.0 12.8 Flocculation reaction, cu eighteen thirty 30.5

table 2
Comparison of the content of biologically active substances and the biological evaluation of extracts of Dolutzar and Luceron Name of indicator Dolucar extract (prototype) Luceron extract The content of vitamin E, mg / ml 7.9 12 The content of vitamin C, mg / ml 2,4 3.57 The content of vitamin B ₂ , mg / ml 19,4 29.4 The content of vitamin B ₁₂ , mg / ml 14.2 23.8 The content of uronic acids, mg / ml 17,2 15.6 Flavonoid content, mg / ml: - according to luteolin 9.8 10,4 - by routine 8.6 12.8 Flocculation reaction, cu fifteen 30.5 The content of heavy metals,% no more 0.01 0.001 Toxicity low toxicity practically non-toxic Wound healing activity, day: - burns 35 thirty - strip skin wounds 23 21 Hepatoprotective activity (mouse survival) 17/20 20/20

Information sources

1. Auth. St. No. 1839723, 1989

2. Patent RU (11) 2145863 C1. 1998 year

3. Avtsin A.P., Zhavoronkov A.A., Rish M.A., Strochkova L.S. Trace elements of man. M., Medicine, 1991

4. Hygienic requirements for safety and nutritional value of food SanPiN 2.3.2.1078-01.

5. Rules for the organization of production and quality control of medicines (GMP) OST 42-510-98.

6. Hygienic requirements for the organization of production and turnover of biologically active food additives (BAA). Sanitary and epidemiological rules and regulations SanPiN 2.3.2.1290-03

7. Sokolov LB Fundamentals of the synthesis of polymers by polycondensation. M., ed. Chemistry, 1979

Claims (2)

1. A biologically active alfalfa extract with high biological activity and immunomodulating properties obtained by treating raw materials with an extractant containing an aqueous solution of salts of trace elements: Fe, Zn, Cu, Mn, Co, Cr, Se, characterized in that the extractant additionally contains salts: Mo, I and F, and salts of trace elements are taken in the following ratio of raw materials, mg / kg: Fe 320.0 Zn 180.0 Cu 45.0 Mn 25.0 From 6.0 Mo 5.0 Cr 4.0 Se 3.0 I 2.0 F 1.0 2. A method of obtaining a biologically active extract from alfalfa, comprising treating the raw material with an extractant, containing an aqueous solution of salts, extracting the mixture, further separation and purification, characterized in that the processing of alfalfa is carried out with an aqueous solution of salts of trace elements: Fe, Zn, Cu, Mn, Co, Mo, Cr, Se, I and F at a temperature of 30-40 ° C for 55-65 min, then the mixture is heated to a temperature of 122-142 ° C and maintained at a pressure of (2-3) × 10 ⁵ Pa for 2 hours, cooled, purified and evaporated at a temperature of 85-95 ° C.