RU2506953C1 - Method for preparing biologically active complex of powdered young sprouts of sea buckthorn - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely a method for preparing a biologically active complex containing serotonin. The method for preparing the biologically active complex containing serotonin involves milling a herbal raw material that is young sprouts of sea buckthorn collected in the early spring and containing 0.9%-1.1% of serotonin in a cutting mill; mixing the milled herbal raw material and ascorbic acid in a blade mixer; carrying out solid-phase mechanochemical treatment in a roll mill in the intensive mechanical environment in the presence of solid edible agents specified in: citric acid - sodium chloride at a dry substance (1:0.5)-(1:1), or acid, alkaline and phosphate-containing agents to prepare a fine powder at particle size 125 mcm.

EFFECT: ensured simplifying the technology, improving the production rate, as well as higher yield of the end product per a unit of raw material, and providing the chemical production safety.

5 cl, 1 dwg, 5 tbl

Description

The invention relates to the field of pharmaceutical and food industries and extends to the production of a biologically active complex - a powder from young shoots of sea buckthorn, containing serotonin as the target component (main active component). and intended for the production of biologically active food additives, products of therapeutic and prophylactic nutrition and as a biologically active component in the production of cosmetic products.

Serotonin (5-hydroxytryptamine, 5-HT) is one of the main neurotransmitters. This substance is also called the hormone of pleasure. Serotonin affects many body functions. In the anterior lobe of the brain with the participation of this hormone, the regions that are responsible for the cognitive process are activated. When this hormone enters the spinal cord, motor function improves and muscle tone increases. But the main function of this hormone in our body is a mood boost that is created in the cerebral cortex. If this substance is not enough in the body, this leads to depression. It also affects emotional stability and susceptibility to stress.

A known method for the production of a biologically active complex containing serotonin from sea buckthorn shoots, in which the bark of young branches is harvested in late spring and dried, and then crushed and prepared a decoction of buckthorn buckthorn bark, for which 1 tbsp. powder pour 2 cups boiling water and boil for 10 minutes. Insist 30 minutes, filter and drink a quarter cup 3-4 times a day (http://q98760q.narod.ru/article_149.htm).

This method is not suitable for industrial production, it does not allow to control, stabilize the properties of the resulting product and to ensure its storage.

Known methods for producing serotonin RU No. 2227140, 2299880, carried out at elevated pressure using various toxic substances.

A known method of producing a biologically active complex containing serotonin, based on a powder from sea buckthorn bark. The extraction of serotonin is carried out by extraction with acidified sulfuric acid with water, followed by passing the extract through a column filled with a cation exchange resin. Then the cation exchange resin is eluted with a hydrochloric acid solution, and the eluate is neutralized with sodium bicarbonate and evaporated to dryness in vacuo under a stream of nitrogen. From the dry residue, serotonin is repeatedly extracted with absolute alcohol and recrystallized (SU No. 137234, prototype).

The disadvantage of this method is the technological complexity and high cost of production, the possibility of contamination of products with toxic substances, as well as the chemical hazard of production.

An object of the invention is to provide an effective method for the production of a biologically active complex containing serotonin. based on the powder from young shoots of sea buckthorn, as well as expanding the arsenal of methods for the production of a biologically active complex containing serotonin, based on the powder from young shoots of sea buckthorn.

The technical result that provides a solution to the problem is to simplify production technology, increase productivity (yield of the target product per unit mass of raw materials) and ensure chemical safety of production.

The essence of the invention lies in the fact that the method of preparing a biologically active complex includes grinding raw materials from sea buckthorn shoots in a knife mill to an average piece size of 0.5 mm, mixing in a stainless steel paddle mixer of crushed plant material and ascorbic acid, and to obtain 100 kg of biologically active powder the complex takes 95.5 kg of ground shoots of sea buckthorn and 5.0 kg of ascorbic acid, as well as the subsequent solid-phase mechanochemical treatment of the mixture by grinding to grow of raw materials in a roller mill under conditions of intense mechanical action in the presence of solid food reagents to obtain a fine powder with a composite particle size of 70-300 microns, and destroyed cell particles as a result of the introduction of particles of a solid reagent, and the composite particles include smaller particles 200- 800 nm of plant material.

Preferably, the raw material is prepared from young shoots collected in early spring up to 400 mm long and with a bottom cut diameter of not more than 5 mm, covered with silver scales and star-shaped hairs with spherical buds, with 2-3 outer scales coated with copper-bronze scales containing extractive substances extracted with 70% alcohol, not less than 12%, humidity not more than 12%, total ash not more than 5%, extraneous matter not more than 1%, and crushed raw materials, in which particles not passing through a sieve with openings of size 0, 5 mm, not more than 10%.

In this case, the quality of the raw material is determined, for which samples are taken - shoots with a diameter of 2-3 mm, which are boiled in a 5% solution of caustic soda for 3-5 minutes. and they prepare a squeezed preparation, which is considered suitable for the preparation of a biologically active complex if xylem elements are visible during microscopic examination: vessels, tracheids, cells of the primary cortex, stellate multicellular hairs, as well as porous and reticular vessels, consisting of short segments and cells epidermis with straight side walls.

In addition, the content of serotonin in the feed is checked by high performance liquid chromatography, the feed is considered suitable for the preparation of a biologically active complex if the content of serotonin is 0.9-1.1%.

As a rule, a powder is obtained with a mass fraction of serotonin of at least 0.8 wt.%, And a mass fraction of ascorbic acid of 5.0-10 wt.%, And with a mass fraction of moisture of 8.0 ± 2.0 wt.%.

An amorphous (crystalline-free) powder is obtained in the form of a mixture of heterogeneous particles of raw materials from yellow-green to brown in color with intercrystalline inclusions, the product obtained is checked for lead, arsenic, cadmium, mercury, hexachlorocyclohexane, DDT and its metabolites, heptachlor and aldrin, the content of which should not exceed (in mg / kg) the values:

Lead 6.0 Arsenic 0.5 Cadmium 1,0 Mercury 0.1 hexachlorocyclohexane (α, β, γ) 0.1 DDT and its metabolites 0.1 heptachlor, aldrin not allowed (less than 0.002)

As a rule, they check the microbiological parameters of the resulting product for microbiological indicators, which should not exceed the values:

KMAFANM, CFU / g, no more 1 × 104 BGKP (coliforms), 0.1 g not allowed E. coli, 1.0 g not allowed S. aureus 1.0 g not allowed Pathogens, including salmonella, in 10 g not allowed Yeast, CFU / g, not more than one hundred Mold, CFU / g, not more than one hundred B. cereus, CFU / g, not more than 200

Figure 1 shows a flowchart of a procedure for the production of a biologically active complex containing serotonin, based on powder from young shoots of sea buckthorn.

A method of manufacturing a biologically active complex containing serotonin is implemented as follows.

All incoming raw materials, auxiliary, packaging materials used for powder production must meet the requirements of pharmacopoeial articles, regulatory or technical documentation, comply with the Unified Sanitary and Epidemiological and Hygienic Requirements for Products Subject to Sanitary and Epidemiological Surveillance (control), and are allowed for use in the production of biologically active additives in the prescribed manner; be accompanied by documents confirming their quality and safety.

Description of plant materials used to implement the proposed method are given in table 1.

Table 1 Description Young shoots of cultivated buckthorn buckthorn bush, Hippophae rhamnoides L., fam. Sucker - Elaeagnaceae External signs Young shoots up to 400 mm long and a lower cut diameter of not more than 5 mm, covered with silver scales and star-shaped hairs with spherical kidneys, with 2-3 outer scales covered with copper-bronze scales. The presence of leaves is allowed (alternate, oblong or linear-lanceolate, short-leaved (almost sessile), single-strand, with a wedge-shaped base, top-silver-gray or silver-green, bottom-brownish-silver or yellowish-white, pubescent). Leaf length 2-7 cm, width 2-8 mm. Microscopy Shoots with a diameter of 2-3 mm are boiled in a 5% solution of caustic soda for 3-5 minutes. A squeezed preparation is prepared in which xylem elements are visible: vessels, tracheids, cells of the primary cortex, stellate multicellular hairs. The vessels are porous and mesh, consisting of short segments. Epidermal cells with straight side walls. Numeric indicators Whole raw materials. Extractives extracted with 70% alcohol, not less than 12%, humidity not more than 12%, total ash no more than 5%, impurities no more than 1%. Ground raw materials. Particles not passing through a sieve with openings of 0.5 mm in size, not more than 10% Quantification of Serotonin in Raw Materials Ground raw materials. Serotonin content 0.9% ± 1.1%. Serotonin content is measured by high performance liquid chromatography (HPLC) on a Millichrom A-02 instrument (EkoNova, Novosibirsk), ProntoSIL 120-5C18AQ sorbent. HClO4 pre-sample (5 μl), LiClO ₄ * 3H ₂ O eluents in HClO ₄ -acetonitrile in gradient mode. The srtnfree technique and a calibration graph of serotonin adipate, linear in concentrations of 0.1-1 g / l, are used.

For the manufacture of powder, ascorbic acid according to FS 42-2668 is used.

Powder production consists of successive stages:

- Training of premises, equipment, personnel

- Preparation of raw materials and containers.

- Grinding plant materials (sea buckthorn shoots)

- Mixing crushed plant materials and ascorbic acid

- Mechanochemical processing of the mixture

- Acceptance, sampling and testing

- Filling and powder packing

To prevent microbial contamination of products, the process is conducted under conditions that limit the possibility of contamination of products with microflora. Processing of industrial premises is carried out with disinfectant solutions using detergents permitted in the established manner for these purposes. Processing of industrial premises is carried out daily and includes cleaning floors, cleaning walls and doors. Control of microbial contamination of air in industrial premises is carried out at least 1 time per week, in accordance with GOST R 52249-2009 "Rules for the production and quality control of medicines" and MU-42-51-4-93.

Inspection of personnel is carried out before starting work daily for lack of disease, clean hands and the availability of clean work clothing for all employees associated with the production, packaging or transportation of finished products.

Before starting work, the apparatchik inspects all technological equipment. Having ascertained the cleanliness and serviceability of the equipment, the apparatchik proceeds to load the components according to the norms of the technological regime on the prepared product.

The operation of the preparation of raw materials includes taking samples of ingredients in accordance with the recipe per one technological operation. Raw materials are weighed on scales in polymer, metal or enameled containers of appropriate capacity.

Packaging preparation consists in visual inspection and rejection of substandard products.

Grinding of sea buckthorn shoots is carried out at a knife mill RM-120. intended for fine crushing of plant materials. The mill is equipped with a plunger for pushing the material into the crushing chamber and the possibility of fixing plastic bags to collect the product. In the process, the crushed material fed through the feed funnel enters the crushing chamber. Grinding (cutting) of the material occurs when particles enter between the blades of the rotor and the housing. Particles of the material experience shear deformations, similar to deformations that occur when shears or a guillotine knife are exposed. For one revolution of the rotor, 12 grinding cycles occur. Basic are 2 modes of operation: with a closed and open crushing chamber. In the first mode, the crushed material (usually long fiber or bulk) is pushed through the vertical channel of the supply funnel with a plunger, the end of which in the lower position overlaps the channel section. In the second mode, the plunger is fixed with a locking screw in the upper part of the vertical channel of the supply funnel and loading is carried out using a pusher through a horizontal trough. The material enters the crushing chamber periodically as the rotor cavities coincide with the loading window of the mill body. Material that does not enter the crushing chamber is thrown into the feed hopper (with the crushing chamber not open). Thus, in the vertical channel of the funnel, bounded from above by the plunger, a "boiling" layer of material is formed. The size of the pieces of sea buckthorn shoots after grinding is not more than 0.5 mm.

Mixing crushed plant materials and ascorbic acid is carried out in a paddle mixer SP-100. In batch-type paddle mixers, the product is usually mixed with radial blades slightly rotated about the axis of rotation. Such a device of the working bodies provides mixing with simultaneous circulation of the product in the trough of the mixer, made of carbon and stainless steel. Pneumatic seals eliminate product spillage. The loading of components (ground sea buckthorn and ascorbic acid) is carried out in accordance with the recipe. The product is unloaded through the discharge bottom without residue.

The mechanochemical processing of the mixture is carried out in a roller mill activator RM-1 developed by the Institute of Chemical Technology and Chemical Technology SB RAS, Novosibirsk. Mill productivity - up to 60 kg of powder product per hour.

The roller mill consists of a sealed housing and a horizontal grinding ring rotating in it, to which two rollers with a diameter of up to 1200 mm are pressed by springs. The source material is fed to the grinding ring and, when it is rotated, is crushed by rollers.

The process of mechanochemical processing consists in grinding plant materials under conditions of intense mechanical action in the presence of solid food reagents (solid-phase mechanochemical processing) of a reagent consisting, for example: of citric acid and sodium chloride in the ratio citric acid - sodium chloride in terms of dry substance (1: 0 , 5) - (1: 1), or acid and alkaline reagents, or phosphate-containing reagent.

To obtain 100 kg of a powder preparation, 95.5 kg of raw materials (sea buckthorn shoots) and 5.0 kg of ascorbic acid are consumed. 0.5 kg of water from sea buckthorn raw materials evaporates during the process of mechanochemical treatment in an activator mill.

The finished product is a fine powder with a particle size of less than 125 microns. Particle cells are destroyed by the incorporation of solid reagent particles. Composite particles include smaller particles of 200-800 nm of plant material, the structure of which is changed as a result of mechanochemical processing.

There is no production waste, with the exception of evaporating water and a small amount of dust, when packaging the product.

As personal protective equipment at work, respirators SHB-1 "Petal", goggles, gloves and protective clothing are used.

Powder production is not harmful to the environment. Waste water after washing technological equipment in industrial wastewater must be neutralized and sent to the city sewer.

According to external (organoleptic) signs, the obtained powder complies with the standards indicated in table 2.

table 2 Name of indicator Indicator value Appearance Amorphous powder Macroscopy Powder - a mixture of heterogeneous particles of raw materials from yellow-green to brown in color with fine crystalline inclusions Color Yellow green to brown Taste and smell Specific, inherent to this product, without musty and foreign smell, with sour taste

According to physico-chemical parameters, the powder meets the requirements specified in table 3.

Table 3 Name of indicator Indicator value Moisture content, % 8.0 ± 2.0 Mass fraction of serotonin,% of mass, not less 0.8 Mass fraction of ascorbic acid, wt.% 5.0 ± 1.0 Mass fraction of total ash,% of mass, no more 5,0 Pass through a sieve with a hole with a diameter of 0.5 mm Full The presence of impurities Not allowed

The content of toxic elements and pesticides does not exceed the permissible norms and levels indicated in table 4.

Table 4 Indicators Permissible levels, mg / kg, not more than Test method Toxic elements lead 6.0 GOST 26932 arsenic 0.5 GOST 26930 cadmium 1,0 GOST 26933 mercury 0.1 GOST 26927 Pesticides hexachlorocyclohexane (α, β, γ) 0.1 MU 2142 DDT and its metabolites 0.1 MU 2142 heptachlor, aldrin not allowed (less than 0.002) MU 2142

According to microbiological indicators, the powder meets the sanitary-epidemiological and hygienic requirements specified in table 5.

Table 5 Indicators Allowed Levels Test method KMAFANM, CFU / g, no more 1 × 10 ⁴ GOST 10444.15 BGKP (coliforms), 0.1 g not allowed GOST R 50474 E. coli, 1.0 g not allowed GOST 30726-001 S. aureus 1.0 g not allowed GOST R 52815 Pathogens, including salmonella, in 10 g not allowed GOST R 50480 Yeast, CFU / g, not more than one hundred GOST 10444.12 Mold, CFU / g, not more than one hundred GOST 10444.12 B. cereus, CFU / g, not more than 200 GOST 10444.8

Acceptance and sampling is carried out in batches. From the combined sample by quartration, an average sample is extracted with a mass of at least 100 g. Analytical samples are isolated from the average sample by the quartet method to determine the authenticity, moisture, fineness and content of impurities, ash and serotonin, ascorbic acid.

Each batch of powder is subjected to acceptance tests for organoleptic, physico-chemical indicators in the laboratory of the manufacturer. The monitoring of the content of toxic elements, pesticides and microbiological indicators is carried out in an accredited laboratory in the manner approved by the manufacturer at least 1 time per quarter and when changing raw materials.

If deviations of the quality indicators of at least one of the indicators from the normalized values of these technical conditions are detected, a repeated analysis is carried out in double the size of the sample taken from the same batch, the results are final and apply to the entire batch.

If negative results are obtained with at least one of the quality indicators during the reanalysis, the entire batch is rejected and sent for recycling.

Serotonin content is measured by high performance liquid chromatography (HPLC) on a Millichrom A-02 instrument (EkoNova. Novosibirsk), ProntoSIL 120-5C18AQ sorbent, HClO4 pre-sample (5 μl). eluents LiClO ₄ * 3H ₂ O in HClO _4- acetonitrile in a gradient mode. The srtnfree technique and a calibration graph of serotonin adipate, linear in concentrations of 0.1-1 g / l, are used. The chromatograph signal, the value of which is proportional to the concentration of the analyte in the eluent stream, is automatically continuously recorded and recorded (for example, on a chart tape).

Note:

The powder is stored in clean, dry, well-ventilated rooms, avoiding direct sunlight and moisture, at a temperature not exceeding 25 ° C. The shelf life of the product in the manufacturer’s packaging is 12 months from the date of manufacture.

The result is a simplification of production technology, increased productivity (yield of the target product per unit mass of raw materials) and a longer storage period, prevention of product pollution with toxic substances, and ensuring chemical safety of production.

Claims (5)

1. A method of preparing a biologically active complex containing serotonin, including grinding plant materials - young shoots of sea buckthorn, collected in early spring, up to 400 mm long and a bottom cut diameter of not more than 5 mm, covered with silver scales and star hairs with spherical kidneys, with 2 -3 outer scales coated with copper-bronze scales; containing extractive substances extracted with 70% alcohol, at least 12%, containing serotonin 0.9-1.1%, moisture content not more than 12%, total ash not more than 5%, impurities not more than 1% - in a knife mill to a piece size of not more than 0.5 mm, mixing the crushed plant material and ascorbic acid in a stainless steel paddle mixer, moreover, to obtain 100 kg of powder of the biologically active complex, 95.5 kg of crushed shoots of sea buckthorn and 5.0 kg of ascorbic acid are taken; then, solid-phase mechanochemical processing of the resulting mixture in a roller mill is carried out under conditions of intense mechanical action in the presence of solid food reagents selected from the group: citric acid - sodium chloride in terms of dry matter (1: 0.5) - (1: 1), or acidic, alkaline and phosphate-containing reagents, to obtain a fine powder with a particle size of less than 125 microns. 2. The method according to claim 1, characterized in that the quality of the raw material is determined, for which samples are taken - shoots with a diameter of 2-3 mm, covered with silver scales and star hairs, with spherical kidneys with 2-3 outer scales coated with copper-bronze flakes that are boiled in a 5% solution of caustic soda for 3-5 minutes and prepare a squeezed preparation that is considered suitable for the preparation of a biologically active complex if xylem elements are visible under microscopic examination: vessels, tracheids, primary cells cortex, stellate multicellular hairs, as well as porous and reticular vessels, consisting of short segments and epidermal cells with straight side walls. 3. The method according to any one of claims 1 and 2, characterized in that a powder is obtained with a mass fraction of serotonin of at least 0.8 wt.% And a mass fraction of ascorbic acid of 5.0-10 wt.%. 4. The method according to any one of claims 1 and 2, characterized in that a powder is obtained with a mass fraction of moisture of 8.0 ± 2.0 wt.%. 5. The method according to any one of claims 1 and 2, characterized in that an amorphous (not having a crystalline structure) powder is obtained in the form of a mixture of heterogeneous raw material particles from yellow-green to brown in color with fine crystalline inclusions.