RU2792928C2 - Method for production of co2 rose root extract - Google Patents

Abstract

FIELD: extraction.

SUBSTANCE: invention relates to a technology of extraction of natural raw materials, namely to methods for the production of CO₂ extract, which can be used in food, perfumery and cosmetics, and pharmaceutical industry. A method for the production of CO₂ rose root extract is proposed, in which plant raw materials in the form of roots of rose root, ground to a particle size of 2-8 mm, are subjected to extraction with a solvent, with a flow rate of 5-20 kg/kg of plant raw materials, as which supercritical fluid CO₂ is used with the mass fraction of ethanol of 3-4%, at a temperature of 31-60°C and pressure of 100-300 bar, for 30-60 min.

EFFECT: invention allows for provision of high efficiency of extraction of rose root due to an increase in the mass fraction of a cosolvent, as which ethanol is used; thermolabile biologically active substances (BAS) in effective amounts (with the general yield of up to 9% of the weight of plant raw materials), in particular salidroside, rodalin, rosavin, rosiridin; reduction in time, temperature, and pressure of extraction; preservation of a composition of biologically active substances in the resulting product.

1 cl, 4 dwg

Description

The invention relates to a technology for the extraction of natural raw materials, and in particular to methods for obtaining a CO2 extract, which can be used in the food, perfume, cosmetic and pharmaceutical industries.

A known method of obtaining CO2-extract, including the preparation of plant materials, followed by CO2-extraction at a pressure of 20-40 MPa for 120 min [see. RF patent No. 2070053, IPC A61K 35/78, publication date 10.12.1996].

The disadvantage of the analogue is the fact that pure CO2 is an inefficient solvent for chemical compounds with a higher polarity.

As the closest analogue, a method for obtaining CO2-extract of ginseng was adopted, including its preparation, followed by extraction with supercritical fluid CO2 with a mass fraction of ethanol of 1.7-3.4% at a liquid flow rate of 250 g/min at a temperature of 31-70°C and pressure of 20-40 MPa for 120 min [see. patent of the Russian Federation No. 2679634, IPC A61K 36/258, A61P 37/00, publication date 02/12/2019].

The disadvantages of the closest analogue are:

- extracting in effective amounts only six heat-labile ginsenosides from Far Eastern ginseng;

- long time, as well as high temperatures and pressures of extraction;

- a small yield of biologically active substances associated with the consumption of a small amount of co-solvent;

- destruction of some thermolabile biologically active components.

The task to be solved by the claimed invention is the development of an effective method for obtaining CO2-extract of Rhodiola rosea.

The technical result, which manifests itself in solving the problem, is expressed as follows:

- ensuring high efficiency of extraction of Rhodiola rosea by increasing the mass fraction of the co-solvent, which is used as ethanol;

- the possibility of extracting thermolabile biologically active substances (BAS) in effective quantities (with a total yield of up to 9% by weight of plant materials), in particular salidroside, rhodalin, rosavin, rosiridin;

- use of available, safe and easily separated ethanol co-solvent;

- reduction of time, reduction of temperature and pressure of extraction;

- preservation of the composition of biologically active substances in the resulting product;

- no hard-to-recycle waste and by-products.

The problem is solved by the fact that in the method for obtaining CO2-extract of Rhodiola rosea, including the preparation of plant materials, followed by extraction with supercritical fluid CO2 with ethanol at a temperature of 31°C and elevated pressure, Rhodiola rosea roots, crushed to a size particles 2-8 mm, extraction is carried out with a solvent with a consumption of 5-20 kg/kg of plant material, which is used as supercritical fluid CO2 with a mass fraction of ethanol 3-4%, at a temperature of 31-60°C and a pressure of 100-300 bar in within 30-60 min.

A comparative analysis of the set of essential features of the proposed technical solution and the set of essential features of the prototype and analogues indicates its compliance with the criterion of "novelty".

In this case, the distinctive features of the claims solve the following functional tasks.

The feature "Rhodiola rosea roots are used as plant material" describes the type of plant material used.

The plant Rhodiola rosea , family Crassulaceae L., Crassulaceae, is widely used in folk medicine and traditional medical systems (Tibetan, Chinese, Korean), the rhizomes and roots of the plant are mainly used [1].

The main medicinal raw material of Rhodiola rosea are rhizomes with roots, which are harvested from the end of flowering until the end of the growing season of the plant.

Rhodiola has been one of the main adaptogenic plants since about the 80s and competes with such well-known adaptogens as the Far Eastern ginseng Panax ginseng and eleutherococcus Eleutherococcus . Adaptogens are a pharmacological group of drugs of natural or synthetic origin that can increase the body's resistance to various adverse environmental factors [2, 3].

The active biologically active substances of R. rosea are tyrosol, salidroside, caffeic acid, gallic acid, methyl gallate, flavonoids (astragallin, campferol, rhodionin, rhodiosin, rhodiolhydin, rhodiolgin, acetyl-rodalgin), tannins of the pyrogallol group. Monoterpenes are represented by rosiridol and its glycoside rosiridin, sterols are represented by β-sitosterol and daucosterol. Cinnamic alcohol glycosides, rosin, rosarin, and rosavin, were isolated from the root of R. rosea .

Data on the content of salidroside and rosavin in R. rosea are numerous and contradictory [4, 5].

Researchers have not yet come to a consensus on the localization and activity of specialized biosynthesis, the nature of seasonal changes in the content of glycosides, and the variability of the accumulation of these substances in wild and cultivated plants [6–8].

The sign "Rhodiola rosea roots, crushed to a particle size of 2-8 mm, are used as plant raw materials" describes the optimal particle sizes of plant materials, which ensure more efficient extraction of biologically active substances.

Signs “extraction is carried out with a solvent with a consumption of 5-20 kg/kg of plant material, which is used as supercritical fluid CO2 with a mass fraction of ethanol 3-4% at a temperature of 31-60°C and a pressure of 100-300 bar for 30-60 min » describe the optimal operating characteristics.

Changing the regime characteristics, both in the direction of decrease and increase, leads to a decrease in the efficiency of extraction of biologically active substances.

The inventive method is carried out as follows.

Pre-prepared vegetable raw materials.

The roots of Rhodiola rosea ( Rhodiola rosea ) are crushed to a particle size of 2-8 mm and loaded into the vessel of the extraction apparatus of supercritical pressure Thar SFC, SN 3526551, USA, already heated with a hot jacket, the temperature is controlled by a thermostat (±1°C). Carbon dioxide (CO2) was compressed using a compressor, the pressure is controlled by a metering valve.

Next, extraction is carried out with a solvent at a flow rate of 5-20 kg/kg of plant material, which is supercritical fluid CO2 with a mass fraction of ethanol of 3-4% at a temperature of 31-60°C and a pressure of 100-300 bar for 30-60 minutes.

The solution of biologically active substances in CO ₂ is subjected to decompression in the separator.

Figure 1 shows the yield of Rosavin depending on pressure and temperature during solvent extraction with a consumption of 15 kg/kg of plant material, which is supercritical fluid CO2 with a mass fraction of ethanol of 3%, for 50 minutes.

Figure 2 shows the yield of Rodalin depending on pressure and temperature during extraction with a solvent at a flow rate of 10 kg/kg of plant material, which is supercritical fluid CO2 with a mass fraction of ethanol of 4%, for 30 minutes.

Figure 3 shows the yield of Salidroside depending on pressure and temperature during solvent extraction with a consumption of 5 kg/kg of plant material, which is supercritical fluid CO2 with a mass fraction of ethanol of 3.5%, for 60 minutes.

Figure 4 shows the yield of Rosiridin depending on pressure and temperature during solvent extraction with a consumption of 20 kg/kg of plant material, which is supercritical fluid CO2 with a mass fraction of ethanol of 3.7%, for 40 minutes.

Multicomponent mixtures were separated by HPLC using a Shimadzu LC-20 Prominence HPLC high-pressure liquid chromatograph (Shimadzu, Japan) equipped with an SPD-20A spectrophotometric detector and a Shodex ODP-40 4E reverse phase column.

The elution gradient program (water - acetonitrile) is as follows: 0-4 min, 100% H ₂ O, 0% acetonitrile (CH ₃ CN); 4-60 min, 100-25% H ₂ O, 0-75% CH ₃ CN; 60-75 min, 25-0% H2O, 75-100% CH ₃ CN; control washing 75-120 min 0% H ₂ O, 100% CH ₃ CN. All HPLC analysis was done with an ESI detector at 230 nm and 330 nm; temperature 17°C. The injection volume was 1 ml.

BAS were identified by tandem mass spectrometry using an amaZon SL mass spectrometer (manufactured by BRUKER DALTONIKS, Germany) equipped with an ESI electrospray ionization source in negative and positive ion modes.

Optimized parameters are obtained as follows: ionization source temperature 70°C; gas flow 4 l/min; gas nebilizer (nebulizer) 7.3 psi; capillary voltage 4500 V; end plate bending voltage 1500 V; fragmentator voltage 280 V; collision energy 60 eV. The mass spectrometer was used in the scan range m/z 100-1.700 for MS and MS/MS. The capture rate was 1 spectrum/s for MS and 2 spectrum/s for MS/MS. Data collection was controlled by the Windows software for BRUKER DALTONIKS.

As can be seen in figure 1, the minimum output of Rosavin at a pressure of 100 bar and a temperature of 31°C is 0.5 mg/g of plant material, and the maximum at a pressure of 300 bar and a temperature of 60°C is 2.34 mg/g of plant material.

As can be seen in figure 2, the minimum output of Rodalin at a pressure of 100 bar and a temperature of 31°C is 0.3 mg/g of plant material, and the maximum at a pressure of 300 bar and a temperature of 60°C is 1.2 mg/g of plant material.

As can be seen in figure 3, the minimum yield of Salidroside at a pressure of 100 bar and a temperature of 31°C is 0.35 mg/g of plant material, and the maximum at a pressure of 300 bar and a temperature of 60°C is 2.27 mg/g of plant material.

As can be seen in figure 4, the minimum output of Rosiridin at a pressure of 100 bar and a temperature of 31°C is 0.8 mg/g of plant material, and the maximum at a pressure of 300 bar and a temperature of 60°C is 3.17 mg/g of plant material.

Based on the data obtained, it can be concluded that an increase in the temperature and time of extraction contributes to a more efficient extraction of biologically active substances from plant materials.

Information sources

1. Saratikov, A.S. Rhodiola rosea - a valuable medicinal plant - golden root. /A.S. Saratikov, E.A. Krasnov. - Tomsk: Ed. Tomsk University. - 1987. - 273 p.

2. Kurkin, V.A. Rhodiola rosea (Golden root): standardization and creation of drugs. Monograph. / Samara, 2015.

3. Saratikov, A.S.; Krasnov, E.A. Rhodiola rosea (golden root). /A.S. Saratikov, E.A. Krasnov. -Tomsk: Ed. Tomsk University. - 2004. - 216 p.

4. Rodin, I.A.; Stavrianidi, A.N.; Brown, A. W.; Shpigun, O.A.; Popik, M.V. Simultaneous determination of salidroside, rosarin, and rosavin in extracts from Rhodiola rosea by high performance liquid chromatography with tandem mass spectrometric detection. / Mass spectrometry. - 2012. - V.9. - No. 1. - S. 61-65.

5. Zhang M. Pharmacokinetic of rosavin in rat plasma with ultra performance LC-MS/MS after intravenous and gavage administration. / M. Zhang, Z. Hu, B. Fang, X. Bao, Z. Xiang, H. Wang // Bioanalysis. - 2019. - Vol. 11(9). - Pp. 837-845.

6. Ioset, K. N., Nyberg, N. T., van, Diermen D., Malnoe, P., Hostettmann, K., Shikov, A. N., and Jaroszewski, J. W. Metabolic profiling of Rhodiola rosea rhizomes by (1) H NMR spectroscopy. // Phytochem. Anal. - 2011. - Vol.22(2). - Pp.158-165.

7. Li, T. and Zhang, H. Identification and comparative determination of rhodionin in traditional tibetan medicinal plants of fourteen Rhodiola species by high-performance liquid chromatography-photodiode array detection and electrospray ionization-mass spectrometry. // Chem. Pharm Bull (Tokyo). - 2008. - Vol. 56(6). - Pp. 807-814.

8. Evstatieva, L., Todorova, M., Antonova, D., and Staneva, J. Chemical composition of the essential oils of Rhodiola rosea L. of three different origins. // Pharmacogn. Mag. - 2010. -Vol. 6(24). - Pp. 256-258.

Claims (1)

A method for producing CO2-extract of Rhodiola rosea, in which plant material in the form of roots of Rhodiola rosea, crushed to a particle size of 2-8 mm, is subjected to extraction with a solvent at a flow rate of 5-20 kg/kg of plant material, which is supercritical fluid CO2 with a mass share of ethanol 3-4%, at a temperature of 31-60°C and a pressure of 100-300 bar for 30-60 minutes.

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