The method of obtaining water-soluble polysaccharides having hepatoprotective activity of leaves of ginseng

 

(57) Abstract:

The invention relates to medicine, namely to the chemical-pharmaceutical industry and relates to a method of obtaining water-soluble polysaccharides from the leaves of ginseng. The leaves of ginseng crushed to particle size of 7 mm is Extracted with water in electro-extraction chamber when the water ratio 1: 20 and the shock waves from pulsed electrical discharge with a duration of 1.2 μs, with the front 5 NS. The number of pulses 1400 series, the amplitude of 20 to 36 kV, the interelectrode gap 1,75 - 2,25 mm shaken out three times with water at 20oC for 8 minutes Extracts unite, concentrate under vacuum, treated three times with a volume of 96% ethanol in the ratio of extractant:ethanol 1:3. Precipitated water-soluble polysaccharides. The precipitate is filtered, washed with ethanol and dried on the glass. The invention improves the product yield, reduce the time of execution of the process to simplify the method and to increase its efficiency. 4 Il., table 4.

The invention relates to medical and pharmaceutical industry and relates to a method of obtaining water-soluble polysaccharides (VRPS) from the leaves of ginseng.

A known method of producing motorstv the systematic study of the aerial parts of Phytolacca American and cultivated ginseng as sources immunotropic agents. Proceedings of the fifth Russian national Congress "Man and medicine". - M., 1998. - S. 423). Crushed raw ginseng leaves, sifted through a sieve with the hole diameter of 7 mm, in an amount of 10 g was filled with 200 ml of distilled water and was extracted by stirring with a magnetic stirrer for 1.5 h at 20oC; the extraction was repeated and raw materials with the extractant was left for 12 h, filtered. The extract dialyzed for 36 h, evaporated and water-soluble polysaccharides was besieged twice by volume of 96% ethanol. The solution decantation, the precipitate was centrifuged, dried in the vacuum advance-drying apparatus. Sediment VRPS weighed. The yield of water-soluble polysaccharides 8,17% by weight of dry raw material.

The disadvantage of this method is its duration and intensity, as well as a small yield of the target product.

Closest to the present invention is the following method of obtaining VRPS from ginseng leaves (Qi-Gao pm, Hiroaki Klyohara, Jong-chol Cyong. and Haruki Yamada. Chemical Properties and Anti - Complementary Activities of Polysaccharide Fractions from Roots and Leaves of Panax ginseng// Planta Medica 55, 1989. - R. 36-38). The leaves of ginseng was extracted three times 3-fold volume of water at t=100oC in a water bath. The resulting meal ginseng leaves three of extras is denalis, concentrated under vacuum to an acceptable amount and deposited 4-fold volume of 95% ethanol. The yield of water-soluble polysaccharides from the leaves of ginseng 6,25% by weight of dry material. The disadvantages of this method are the low yield of the target product and the duration of the method.

The objective of the invention: improved output VCS, have hepatoprotective activity, simplification of the method of their derivation, reduction of its duration, the recycling efficiency of ginseng by using ginseng leaves.

This objective is achieved in that in the method of obtaining WRPS, including three-time extraction of the leaves of ginseng, combining the extracts, their concentration under vacuum, the deposition of water-soluble polysaccharides 3-fold volume of 96% ethanol followed by filtration and drying, extraction is carried out in electro-extraction chamber with water at a water ratio of 1: 20 and the shock waves from pulsed electrical discharges in the breakdown voltage of 27 kV, duration of 1.2 μs with the front 5 of the national Assembly, is installed in the chamber interelectrode gap of 2 mm, the processing of raw materials is carried out three times, 1400 pulses in series, each for 8 minutes.

operating the camera with a volume of 500 ml, made of low pressure polyethylene, with adjustable interelectrode gap.

In Fig. 1. depicts the extraction chamber 3 with the electrodes 1, 2 in which was placed a portion of the raw material 4 and poured water in a certain ratio with dry raw material. The electrodes 1, 2 of the extraction chamber was fed a series of pulses with a voltage of from 20 to 36 kV with a pulse energy of from 20 to 64 j respectively.

The energy of the pulse voltage is calculated according to the formula and is selected from a ratio of 20 W64 J.,

W=CUn2/2,

where Un- breakdown voltage;

With the capacity of the generator in shock.

The rationale for the choice of parameters given in table. 1-3.

Increase the time of processing of raw materials leads to clouding of the solution due to the overgrinding of raw materials, difficulty filtering, and also degrades the quality of the extract. The maximum yield of extractives is observed when the value of the breakdown voltage of 27 kV and processing of raw materials within 24 minutes (3 series 1400 pulses). On table. 1, figs. 2 shows the output VCS when processing without change of extractant (series a) and three-time processing with the change of the extractant (series In the second series after the change of the extractant, series C - tree, what is more effective is a three-time processing ginseng leaves. The total yield of the target product increased almost twice compared to the usual series of 4200 pulses in a row, when the curve of the percentage of WRPS in the extract reaches saturation at 1400 pulses.

There is also the optimal value of the interelectrode gap (IEP), which is typical for this type of feedstock (see table. 2, figs. 3). Weight allocated VCS to a large extent depends on the size of the map in the investigated range of values increases in proportion to its size due to the increase of the volume of the mixture exposed to shock waves. The restriction map was mainly due to the capabilities of the high-voltage pulse generator. Once its value reaches the value when it is impossible to establish a stable regime of discharge, the process ceases to be effective. With the growth of the IEP increases the length of the discharge channel in the environment and the volume of processed raw materials, exposed to pulsed loads. This process is characterized by a higher energy, because it takes place at the increased value of the breakdown voltage.

The most productive, with maximal field E kV/see When increasing the breakdown voltage is higher than the optimal time lag of development of the discharge is reduced, which is equivalent to the reduction in the duration of the current impulse voltage, and thus the current pulse. This leads to changes in the parameters of the discharge channel and the change of the characteristics affecting raw material factors (see tab. 3, Fig. 4). In table. 3 and in Fig. 4 shows a single processing 1400 pulses (series a) and three-time processing on 1400 pulses in the series (series b).

While processing the raw materials with application of pulsed electrical discharge grinding of raw materials to a particle size of 1 mm, which contributes to the rupture of the cells, increasing the surface of the raw material, in contact with the extractant and contributes to the intensification of the process of extraction of biologically active substances from plant pores of the capillaries. High-power pulsed electrical discharge in the liquid initiates around the discharge channel number of events causing the electrohydraulic effect. In the processes of extraction and homogenization of the most fully used energy diverging fluid and the energy of the shock wave. What if this destruction of plant cells favors ek is rashisa in intracellular juice. This may explain the increase in the yield of the target product. Thus, if in a known way, a certain part of VRPS remains uncovered, the proposed method of selection of the target product is almost entirely due to the hard mode of extraction.

When using a pulsed electric discharge is heated extraction mixture to a temperature of about 30-40oC, what is the optimum temperature for extraction WRPS.

The highest yield of polysaccharides occurs when the water ratio 1:20, because at this rate observed maximum extraction VRPS from ginseng leaves.

The method is illustrated by the following examples.

Example 1. The method of obtaining water-soluble polysaccharides that have hepatoprotective activity, is as follows: is three-fold extraction with the change of the extractant for 8 minutes, after which water extraction unite, concentrate under vacuum, the precipitated 3-fold volume of 96% ethanol, the precipitate was filtered and subjected to drying in a vacuum drying Cabinet. The extraction is carried out in electro-extraction chamber at t= 20oC, the water ratio 1:20 and the impact of daryaganj 27 kV, the electrode gap 2.00 mm, the number of pulses 1400 in series with a repetition rate of 3 pulses/s, three times for 8 minutes, and the precipitation of the polysaccharides are 96% ethanol.

Output VBS was 16.8% compared to the absolutely dry weight of raw materials.

Example 2. 5 g of leaves of ginseng is loaded into the extraction chamber, poured 100 ml of water at room temperature. Raw materials processed by pulsed discharges with a duration of 1.2 μs with front 5 NS, the breakdown voltage of 27 kV, the number of pulses 1400 series, the interelectrode gap of 1.75 mm After three times of extraction with the change of extractant water extraction unite, concentrate under vacuum, the precipitated 3-fold volume of 96% ethanol, dialist, filtered, the precipitate washed with 96%ethanol and subjected to drying in a vacuum drying Cabinet.

Output VBS amounted to 14.3% compared to the absolutely dry weight of raw materials.

Example 3. 5 g of leaves of ginseng, passing through a sieve with a diameter of 3 mm, is loaded into the extraction chamber, poured 100 ml of water. Raw materials processed by pulsed discharges with a duration of 1.2 μs with front 5 NS, the breakdown voltage of 27 kV, the number of pulses 1400 in series with a frequency of 3 CPS, the interelectrode gap t and subjected to drying.

Output VBS was 7.9% compared to the absolutely dry weight of raw materials.

Water-soluble polysaccharide complex obtained from the leaves of ginseng, were tested on a possible hepatoprotective activity. As a pathological model of liver disease were studied three times a day oral administration of white rats hepatotoxic poison - carbon tetrachloride (CCl4) at a dose of 0.3 mg per 100 g weight of the animal. In this study drugs were administered orally for 10 days daily white rats at a dose of 75 mg/kg pre -, then together with hepatoprotector.

We investigated the following series of animals (males, weight 200-250 g).

The first series

White rats, which were injected aqueous solution ITRS ginseng leaves in a volume of 1 ml from the calculation of the dose of 25 mg/kg weight of the animal. Introduction produced daily for 13 days, last 6 days introduction combined with the simultaneous application through day hepatotoxin - 50%-aqueous solution of carbon tetrachloride in vaseline oil at the rate of 0.3 ml per 100 g of body weight of the animal.

The second series of control

Experimental white rats were administered 50% solution of carbon tetrachloride at the rate of 0.3 ml of carbon tetrachloride per 100 g of body weight. Varma), not exposed to any effect.

Fourth series

Experimental white rats were injected in the mode similar to the first series, the officinal solution of the drug "Zelibor" (Mashkovsky M. D. Medicines. T. 1 - M, Medicine, 1992. - S. 512) at a dose of 25 mg/kg weight of the animal. The number of animals in each series was 10.

The introduction of the drug, as well as solution hepatotoxin was carried out using a syringe with a needle on the end with the olive, which was administered per os in the esophagus of the animal.

After the last injection the animals were killed by means of decapetala. In obtained after slaughter, the blood serum was determined by the following tests are considered to be informative in toxic liver injury:

1) was determined by the activity of aminotransferases in serum (Reitman, Frankel, 1957 (Kolb Century BC, Kamyshnikov B. C. Handbook of clinical chemistry. - Minsk, Belarus, 1982. - S. 90-92));

2) was determined by alkaline phosphatase activity in serum by hydrolysis;

3) determined the content of total cholesterol in serum, based on the reaction of the Lieberman-Burchard method ilca (Kolb Century BC, Kamyshnikov B. C. Handbook of clinical chemistry. - Minsk, Belarus, 1982. - S. 90-92)), data sacrificial activity in the model of acute toxic hepatitis, compared with the official activity of the drug "Zelibor". Hepatoprotective activity was combined with an equally high gipotermiceski action (growth inhibition activity of alanine aminotransferase observed in the model). These properties suggest that water-soluble polysaccharides derived from the leaves of ginseng, can be used to create effective hepatoprotective drug.

Thus, the proposed method of producing water-soluble polysaccharides from the aerial parts of ginseng differs from the prototype a high yield of biologically active substances in the final product due to the use of electro-processing of raw materials and a significant reduction in processing time ginseng leaves.

Literature

1. Yakovlev, S., D. Muravyev A., Vasilyev, O. N., Muravyova T. I. Chemical study of the aerial parts of Phytolacca American and cultivated ginseng as sources ionotropic agents. Proceedings of the fifth Russian national Congress "Man and medicine" - M., 1998. - S. 423.

2. Qi-Gao pm, Hiroaki Klyohara, Jong-chol Cyong. and Haruki Yamada. Chemical Properties and Anti-Complementary Activities of Polysaccharide Fractions from Roots and Leaves of Panax ginseng// Planta Medica. Journal of analytical chemistry, 1994. - So 49. - Vol. 11. - S. 1205-1206.

4. Barsky, A. C. a Study of the extraction of plant materials and extraction of water-soluble substances using pulsed electric discharges. Abstract of thesis Cand. technology. Sciences. - Tomsk, 1998. - 28 S.

5. Molchanov, A. A. Intensive processing of crude drugs. - M.: Nauka, 1981. - 75 S.

6. Mashkovsky M. D. Medicines. So 1. - M.: Medicine, 1992. - S. 512.

7. Kolb Century BC, Kamyshnikov B. C. Handbook of clinical chemistry. - Minsk, Belarus, 1982. - S. 90 - 92.

The method of obtaining water-soluble polysaccharides having hepatoprotective activity of leaves of ginseng, including three-time extraction of plant materials with water, combining the extracts, their concentration under vacuum, the deposition of polysaccharides threefold volume of 96% ethanol followed by filtration and drying of the target product, characterized in that the extraction is carried out in electro-extraction chamber at 20oC, the water ratio 1 : 20 and the shock waves from pulsed electrical discharge with a duration of 1.2 μs, with the front 5 NS, when the value of the breakdown voltage of 27 kV, electrode gap the Idov are 96% ethanol.

 

Same patents:

The invention relates to medicine, namely to surgery, and can be used for treatment of suppurative cholangitis

The invention relates to the field of medicine and is suitable for the treatment of cholecystitis, hepatitis, as well as urinary tract infections and gastroenteritis

The invention relates to medicine, namely to infectious diseases, and can be used for the treatment of acute and chronic viral hepatitis In children

The invention relates to pharmaceutical compositions containing as active ingredient a derivative of triazine following General formula I or its MES, or its salt and a derivative of triazine
The invention relates to medicine, namely to the creation of collections of medicinal raw material that is used as a cholagogue
The invention relates to medicine, namely to the creation of collections of plant materials that can be used in the treatment of acute and chronic inflammatory lung diseases, diseases of the upper respiratory tract, in the treatment of bronchial asthma
The invention relates to medicine, namely to create charges of crude drugs for the treatment of diseases of the gastrointestinal tract

The invention relates to medicine, namely to methods of using dietary supplements on the basis of the Jerusalem artichoke as a therapeutic and prophylactic means of a wide spectrum of pharmacological action

The invention relates to medicine, namely to the medicines of cough and methods for their preparation

The invention relates to the field of medicine and relates to a drug sedative action
The invention relates to chemical-pharmaceutical industry, in particular to methods for alkaloid platifillina from vegetable raw materials

The invention relates to medicine, in particular to pharmacology, and can be applied in proctology for the treatment of hemorrhoids and fissures of the rectum

The invention relates to medicine, namely to the pharmaceutical industry, and for candles, which can find use for the treatment of external and internal hemorrhoids and rectal fissures

The invention relates to medicine, pharmacology, veterinary medicine and biology

Antiseptic drug // 2151599
The invention relates to medicine, in particular to pharmacology

The invention relates to medicine, namely the pharmaceutical industry and relates to a medicinal product for the treatment of proctologic diseases

The invention relates to medicine, namely to the chemical-pharmaceutical industry and relates to a method of obtaining pectic polysaccharides from vegetable raw materials
Up!