Method for production of powdered preparations containing serotonin from non-fruit sea-buckthorn parts

FIELD: food industry.

SUBSTANCE: method involves raw material milling, extraction of useful substances, filtering and drying. The method characterisation is as follows: extraction is performed in an ultrasonic bath with a food acid with concentration not in excess of 5% in conversion to adipic acid at a temperature of 20-40C during 15-30 min while drying is performed by high-temperature spraying method at a temperature of 70-100C in a flow of extract ensuring production of a powered product. The food acid may be represented by adipic, orthophosphoric, malic, ascorbic, amber or acetic acid.

EFFECT: method enables production of powered preparations of non-fruit sea-buckthorn parts with high serotonin content.

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sabreena relates to pharmaceutical and food industry, in particular, to the production of compositions of biologically active substances, which can be used as a dietary Supplement. More specifically, a new method for producing powder of drugs on the basis of serotonin from vegetable raw materials - narodowych parts of sea buckthorn.

The neurotransmitter serotonin is involved in various physiological aspects of human life and animals. Serotonin is a stimulator of smooth muscle motility and secretory activity of the gastrointestinal tract; increases capillary stability, participates in the regulation of the cycle of sleep - Wake and other important processes [Mohammad-Zadeh L. F., Moses L., Gwaltney-Brant S. M. Serotonin: a review. // Journal of veterinary pharmacology and therapeutics. 2008. Vol.31, No. 3. P. 187-199., Berger M., Gray J. a, Roth B. L. The expanded biology of serotonin. // Annual review of medicine. 2009. Vol.60. P. 355-366]. It is known that the levels of neurotransmitters in General and serotonin in particular largely determines the behavioral features of the animal or human, his mood, tone and social behavior (McGuire, M. T., Troishi A., 1998. Darwinian psychiatry, New York: Oxford University Press, J. H. Turner, Handbook of Social theory//Springer, 2000, 745p), which was first shown in primates (R. D. Masters, M. T. McGuire, The Neurotransmitter Revolution: Serotonin, Social Behavior, and the Law// Southern Illinois University, 1994, 285 p.).

Well known m the methods of qualitative and quantitative analysis of serotonin in various biological objects, both animal and vegetable. The most popular are the chromatography in various modifications (HPLC, capillary chromatography, gas, often combined with methods of mass spectroscopy) and methods based on UV spectra or spectra of luminescence (Saracino, M. A. et al. Chromatographic analysis of serotonin, 5-hydroxyindolacetic acid and homovanillic acid in dried blood spots and platelet poor and rich plasma samples. // Journal of chromatography. A. Elsevier B. V, 2010. Vol.1217, No. 29. P. 4808-4814., Israel M. A chemiluminescent serotonin assay // Neurochemistry International. 2003. Vol.42, No. 3. P. 215-220). The HPLC method has been established increased serotonin levels in narodowych parts of sea buckthorn - young twigs and shoots, and it is in their young crust (I. Brad., G. A. Vlasceanu, I. L. Brad, S. T. Manea, Characterization of sea buckthorn fruits and copses in terms of serotonin and microelements// Innovative Romanian Food Biotechnology, Vol.1, p. 24-30, 2007).

Typically, obtaining herbal products includes grinding and extraction of raw materials, for example, RF application No. 98122555/14.10.12,1998, in which the inventive tool is obtained by extraction of plant raw water-carbohydrate alcohol extractant.

The disadvantage of this method is to obtain the liquid-phase product, which causes the degradation of many biologically active compounds, including serotonin.

Not found in literature mentions about getting dietary supplements of plant materials containing significant amounts of serotonin is a (more than 2% by weight) in principle, and using high-temperature spray drying in particular.

Commercial preparations, "increasing serotonin levels, based mainly on blocking pathways of degradation and conclusion of serotonin (lock MAO and selective inhibitors of serotonin reuptake) and not directly contain serotonin (MacGillivray S. et al. Efficacy and tolerability of selective serotonin reuptake inhibitors compared with tricyclic antidepressants in depression treated in primary care: systematic review and meta-analysis. // BMJ (Clinical research ed.). 2003. Vol.326, No. 7397. P. 1014). Experimentally it was found that production of powders containing serotonin, from an extract of plant materials cannot classical method - evaporation of the solvent, because serotonin is degraded in solution leaving the liquid phase, especially at elevated temperatures.

The objective of the invention is to develop a new method for producing powder of drugs from narodowych parts of sea buckthorn with serotonin more than 2%.

The problem is solved by the creation of a method that includes the grinding of raw materials, the extraction of useful substances, filtration and drying, in which according to the invention the extraction is carried out in an ultrasonic bath with an aqueous solution of edible acid in concentration of not more than 5% in terms of adipic acid at a temperature of 20-40C for 15-30 minutes, and the drying is produced by the method of h is atemperature sputtering at a temperature of 70-100C and the extract stream, providing a powder product.

As a food acid can be used adipic, phosphoric, malic, ascorbic, succinic, and acetic acid.

Example 1

A method for producing powder of drugs from narodowych parts of sea buckthorn with serotonin more than 1% on the basis of high-temperature spray drying consists of the following stages: preparation of plant material, extraction of serotonin and other water-soluble substances from the wood of the variance of sea-buckthorn, filtering the extract, the use of high temperature spray drying

Preparation of plant materials

The original plant material - nalogovye parts of sea buckthorn (young twigs, shoots and leaves) was ground in a knife mill to a particle size that provides the specific surface area of not less than 0.1 cm2/g and dried in the air. The resulting powder was used in further experiments. The humidity of the powder was 8-10%, the average content of serotonin 1 0,1%.

Extraction of serotonin and other water-soluble substances from the wood of the variance of sea buckthorn

Obtained at the stage of preparation of herbal raw material powder was subjected to exhaustive extraction. Exhaustive extraction of serotonin and other water-soluble substances was carried out in an aqueous solution of adipic acid with what ispolzovaniem different water ratios in the range of 1:5-1:40 (solvent - 5% aqueous solution of adipic acid) in an ultrasonic bath WiseClean WUC-A03H (frequency ULTRASONIC 40 kHz) for 15-30 min at a temperature of 20-40C. experiments have shown that the principal effect of shape, size and material (glass/plastic) reactor for the process of exhaustive extraction no.

Exhaustive extraction of serotonin is ensured by the application of ultrasound for at least 15 minutes, while using ultrasound over 30 min degradation of serotonin in the solution. Temperature range 20-40C is selected so that the extraction was carried out in the minimum time interval (the higher the temperature, the faster the extraction), but was not observed degradation of serotonin and reducing its content in the liquid phase in the extraction process.

Obtaining extract

To extract the mixture was filtered sequentially through a gauze filter, "red ribbon", "blue ribbon", "membrane filter" (50 microns).

Serotonin identified in the extract by gas chromatography-mass spectrometry (HPLC-MS). In the HPLC-MS analysis consisted of a liquid chromatograph Agilent 1200 (with diode-array detector) and a hybrid quadrupole-time-of-flight mass spectrometer (micrOTOF-Q (firm Bruker). Column: Bond SB-Aq, 2.1150 mm, 3.5 micron. Eluent: 2%HCOOH-ACN(0-5%, 0-10'; 5-100%, 10-20'). The flow rate of 0.2 ml/min UV det is sterowanie was conducted at three wavelengths: 260/8, 340/32 and 276/8 nm. In addition, continued every second of available UV spectra (150 spectra per minute) in the range 230-500 nm for comparison with the UV-spectra of identified compounds. The solvent 1-0.01 M solution of chloric acid in water, solvent 2 - methanol. The operating parameters of the mass detection. The method of ionization, electrostatic spraying at atmospheric pressure (API-ES). Scan positive/negative ions in the range m/z=100-1000. The gas flow desiccant (nitrogen) 8 l/min, 250C, pressure spray 1.6 bar.

The use of high temperature spray drying

Obtaining a powder of water-soluble drugs was carried out on the installation of spray drying "BUCHI Mini Spray Dryer B-290". The temperature of 70-100C. the Flow of the extract to 1 ml/min, limited technical characteristics of the device.

Temperature range is limited to the instrument characteristics and the characteristics of the solvent. The range of water ratio is justified by the fact that when using the mass ratio of plant raw water is less than 1:5 having difficulty with separation of the liquid and solid phases by filtration due to the swelling of plant material, when using a ratio greater than 1:40 are having difficulty in accurate determination of serotonin in the extract due to the strong dilution. Experimentally until the ANO, for extraction can be used other used in the food industry acids (phosphoric acid, malic acid, ascorbic acid, succinic acid, acetic acid) at a concentration of 5 wt.% in terms of adipic acid, given pKa, solubility, and other individual characteristics of acids.

As in example 1 were carried out the experiments with the above acids, the results are shown in table 1

Table 1
The results of extraction of serotonin various food acids in the temperature range 20-50C for 20 min in an ultrasonic bath
Food acidThe temperature of extraction, CThe serotonin content in the extract in terms of dry weight of plant material, %% extracted serotonin from theoretically possible (0,95% in terms of dry weight)The result of extraction
Adipic200,930,0598%5% +
300,920,0597%5%+
400,940,0599%5%+
500,800,0584%5%-
Orthophosphoric200,950,05100%5%+
300,950,05100%5%+
400,940,0599%5%+
500,760,0580%5%-
Apple 200,920,0597%5%+
300,920,0597%5%+
400,940,0599%5%+
00,830,0587%5%-
Ascorbic200,930,0598%5%+
300,910,0596%5%+
400,930,0598%5%+
500,810,0585%5% -
Amber200,920,0597%5%+
300,940,0599%5%+
400,910,0596%5%+
500,830,0587%5%-
Acetic200,940,0599%5%+
300,940,0599%5%+
400,950,05100%5%+
500,750,0579%5%-

From table 1 we can conclude that the degradation of serotonin when using temperatures above 40C to reduce its content in the extract. It is experimentally shown that the use of temperatures below 20C does not improve the results, but leads to an increase in time of extraction.

Analysis of the serotonin content in the obtained powder samples were carried out as follows: a weighed sample of the obtained powder of the drug in an amount of 0.1 g was dissolved in 1 ml of distilled water. We determined the content of serotonin in the solution and were converted to mass percent in the powdered drug.

Determination of serotonin in solution was carried out by HPLC on device "Milligram a-02". For calculations of serotonin was used calibration graph constructed on the basis of solutions of a pharmaceutical product "adipate serotonin" (CJSC "LORRE M"). The graph is linear in the concentration range of 0.01-5 g/l For the qualitative assessment was used parameters such as retention time and spectral relations at five wavelengths (232 nm, 250 nm, 260 nm, 290 nm, 330 nm).

In the powdered drug serotonin content of not less than 2 mass%. Thus, C is due to removal at the stage of extraction of the major water-insoluble components, such as cellulose, hemicellulose, lignin, there is an increase in serotonin in the final powder product is more than 2 times compared to the initial content in the plant material.

1. A method for producing powder preparations containing serotonin, narodowych parts of sea buckthorn, which includes the grinding of raw materials, the extraction of useful substances, filtration and drying, characterized in that the extraction is carried out in an ultrasonic bath with an aqueous solution of edible acid in concentration of not more than 5% in terms of adipic acid at a temperature of 20-40C for 15-30 min, and the drying is produced by the method of high-temperature sputtering at a temperature of 70-100C and the extract stream, providing a powder product.

2. The method according to p. 1, characterized in that as a food acid can be used adipic, phosphoric, malic, ascorbic, succinic, acetic acid.



 

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