RU2423991C2 - Antioxidant hepatoprotector and its production method - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to chemical-pharmaceutical industry, namely to application of an extract of an elevated part of scabiose centaury (Centaurea scabiosa L.) as an antioxidant hepatoprotector. A method of preparing the antioxidant hepatoprotector consists in grinding the elevated part of scabiose centaury (Centaurea scabiosa L.) to particle size 2-4 mm followed by three-fold extraction in 70% ethanol at temperature 75-80°C for 1-1.5 hours in the raw material to extractant relation 1:15-1:20, evaporation to dryness of the prepared extract.

EFFECT: application of the extract of the elevated part of scabiose centaury (Centaurea scabiosa L) allows extending the range of herbal antioxidant hepatoprotectors, promotes normalising the antioxidant activity in hepatic tissue, regression of the main biochemical values in hepatitis.

2 cl, 20 ex, 9 tbl

Description

The invention relates to the pharmaceutical industry and relates to methods for producing hepatoprotective, having antioxidant effect from plant materials.

Among the variety of drugs used in the complex treatment of diseases of the liver and biliary tract, a relatively small group of drugs that have a selective effect on the liver - hepatoprotectors - is distinguished. In our country, 3-4 drugs are used as hepatoprotectors, most of which are imported, which to some extent limits their availability for clinical use. All this makes it necessary to search for new domestic effective hepatoprotective agents, in particular, herbal origin.

Closest to the proposed is a method for producing drugs (carlsil, silibor) with a hepatoprotective effect, which consists in extracting the fruits of milk thistle with spotted 80% ethanol by repercolation followed by evaporation, treatment with 50% ethanol, degreasing with carbon tetrachloride and treatment with chloroform-ethanol mixture, followed by drying [one].

The known method has a number of significant drawbacks: firstly, extraction with 80% ethanol is carried out at elevated temperatures, secondly, toxic solvents (chloroform and carbon tetrachloride) are used, thirdly, a shortage of raw materials in the Russian Federation, a small biomass of fruits and the use of introduced raw materials . Thus, obtaining extract from the fruits of milk thistle is a complex and lengthy process from a technological point of view. In addition, the therapeutic efficacy of the prototype is not high enough and somewhat inferior to the claimed tool in terms of severity of hepatoprotective and antioxidant properties.

A new technical problem is the expansion of the arsenal of hepatoprotective agents with antioxidant action obtained from plant materials with higher activity and lower toxicity, as well as methods for their preparation.

The problem is solved by using an extract of 70% ethanol from the aerial part of the rough cornflower (Centaurea scabiosa (L.) of the family Asteraceae) as a hepatoprotective agent with antioxidant effect. The method of obtaining it consists in the extraction of the aerial part with 70% ethanol at a degree of grinding of raw materials of 2-4 mm, at a temperature of 75-80 ° C for 1-1.5 hours, with a ratio of raw material-extractant 1: 15-1: 20 and multiplicity extraction 3, followed by removal of the extractant to obtain a dry extract.

Distinctive features showed new properties in the claimed combination - it was first established that an extract on 70% ethanol was used as a hepatoprotective agent with antioxidant effect, and its preparation consists in extracting plant materials, and the aerial part of a rough cornflower, crushed to particle size, was subjected to extraction. 2-4 mm, 70% ethanol at a temperature of 75-80 ° C for 1-1.5 hours, with a ratio of raw materials-extractant 1: 15-1: 20 and extraction ratio 3, followed by evaporation to dryness Acquiring extract that enhances hepatoprotective and antioxidative activity of the desired product.

The method is carried out as follows.

Shredded air-dried leaves, flowers and stems of a cornflower rough collected in the flowering phase, pour 70% ethanol and extracted at a temperature of 75-80 ° C for 1-1.5 hours at a ratio of raw material extractant 1: 15-1: 20 , then cooled to room temperature, the resulting extract is separated from the feed by filtering. The separated extract is purified from mechanical impurities by filtration. After the first extraction, the coarse cornflower meal is poured with 70% ethanol and extracted at a temperature of 75-80 ° C for 1-1.5 hours at a ratio of raw material-extractant 1: 15-1: 20, then cooled to room temperature, the obtained extract is separated from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration. Next, the meal after the second extraction is poured with 70% ethanol and extracted at a temperature of 75-80 ° C for 1-1.5 hours at a ratio of raw material-extractant 1: 15-1: 20, then cooled to room temperature, the resulting extract is separated from the raw material by filtering. The separated extract is purified from mechanical impurities by filtration. The extracts were combined, the solvent was removed in vacuo and the residue was dried.

The dry extract is a brown powder with a peculiar smell. The main biologically active substances exhibiting hepatoprotective and antioxidant properties in extracts of rough cornflower are flavonoids (PL) and phenolcarboxylic acids (PCA). Quantitative determination of the PL content was carried out spectrophotometrically using GSO rutin based on a complexing reaction with aluminum chloride [7]. The quantitative content of PCA was determined by extraction spectrophotometry based on the selective extraction of PCA with ethyl acetate from aqueous extraction at a certain pH value [6].

The quantitative content of PL in the dry extract is 2.85% and FCC is 1.60%.

Pharmacological tests

The hepatoprotective and antioxidant activity of a dry extract of cornflower, obtained by extraction with 70% ethanol, was studied in vivo in a model of acute toxic liver damage [2, 3]. The experiments were performed on white outbred male rats weighing 180-220 g. Acute hepatitis was caused by intragastric administration of a 50% oil solution of carbon tetrachloride in a dose of 1 ml / kg of animal weight for four days. The dry extract of a cornflower rough in the form of a starch suspension was administered to animals intragastrically at doses of 100, 200 and 400 mg / kg of weight 1 time per day for six days. As a comparison drug, the herbal medicinal product Karsil was used, which was administered at a dose of 100 mg / kg [2].

The hepatoprotective effect of the studied extract was evaluated by its effect on the biochemical parameters of the liver and blood serum: the activity of alanine aminotransferase (ALT), aspartate aminotransferase (ACT), the content of bilirubin (total and conjugated), thymol test and alkaline phosphatase (ALP) [4].

In case of damage to biological structures, an important role is given to lipids, phospholipids, as well as lipid peroxidation (LPO) processes, the initiation of which leads to significant transformations of the cell membrane structures, which underlies the pathogenesis of many diseases (in particular, damage to liver cells) [3, 5 ].

Based on this, the study of the effect of rough cornflower extract on LPO processes was evaluated by the content of diene conjugates (DC) and malondialdehyde (MDA) in the liver and blood serum homogenate, as well as the activity of red blood cell catalase and antiradical activity (ARA) of lipid-soluble antioxidants of cell membranes hepatocytes.

Lipid metabolism was judged by the total serum and liver serum total lipids (OL) and phospholipids (OFL).

Statistical processing of the results was carried out using the StatSoft Statistica v6.0 software package with determination of the arithmetic mean (M) and its error (m), as well as using the nonparametric Wilcoxon criterion.

As shown by biochemical studies, in animals treated with carbon tetrachloride for four days, the phenomenon of cytolysis is observed, while the activity of ALT and ACT enzymes increased 2 and 1.7 times, respectively, 1.9 times the concentration of total bilirubin and 4.5 times - conjugated, alkaline phosphatase - 1.43 times. The intensification of LPO processes was noted due to an increase in the content of diene conjugates by 1.5 and 2.3 times and the level of malondialdehyde by 1.2 and 1.3 times in liver tissue and blood serum, respectively, due to a decrease in antioxidant activity of lipid-soluble antioxidants of cell membranes by 66% and catalase activity by 45 % compared with the intact group of animals.

During therapy with the extract from the aerial part of the cornflower rough at doses of 100 and 200 mg / kg of animal weight, similar to the administration of the drug Karsil for six days, regression of the main biochemical parameters in the blood serum of rats with hepatitis is observed, and normalization of antioxidant activity in the liver tissue is revealed, which, in turn, helps to reduce the level of diene conjugates and malondialdehyde (Tables 2, 3).

Examples of specific performance of the method of obtaining the extract from the aerial part of the cornflower rough

Raw materials were collected during the flowering phase in the vicinity of Kemerovo and used in all experiments.

Example 1

100.0 g of raw material, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 2 l of 70% ethanol and heat at 80 ° C for 1 hour, then cool to room temperature, separate obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration. Next, the extraction of the processed raw materials is repeated twice similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.85% and FCC is 1.60%.

The obtained extract on 70% ethanol in the form of a suspension of 1% starch mucus was administered for six days at doses of 100, 200, 400 mg / kg of rat mass with toxic liver damage with tetrachloromethane intragastrically, while the extract in doses of 100 and 200 mg / kg showed the highest activity (Tables 1, 2). According to the results of biochemical studies, the ALAT content of the blood serum of the studied animals when using the extract at 70% ethanol at doses of 100 and 200 mg / kg against the background of toxic liver damage was 10.39 and 11.37 U / L, ASAT - 12.68 and 11 , 38 U / L, total bilirubin content - 40.07 and 35.5 μmol / L, alkaline phosphatase - 134.17 and 116.76 μmol / (min · l), DC in the liver homogenate 85.0 and 85.75 IE / g, malondialdehyde - 8.91 and 19.52 μmol / g, catalase - 153.76 and 153.76 μmol / l · s, ARA - 22.68 and 22.9%, respectively (table 1, 2). The data of lipid metabolism in rats during therapy with extract of cornflower rough at doses of 100 and 200 mg / kg amounted to the following values: the content of OL in serum and liver was 43.48 and 32.88 mg / g and 3.38 and 2.93 g / l, respectively, and the content of OFL - 12.61 and 11.15 mg / g and 0.29 and 0.44 g / l, respectively (table 3). When compared with carlsilum, the extract of cornflower rough with CCI ₄ hepatitis in rats:

1. more effectively reduces the content of ALT, ACT and alkaline phosphatase in the blood serum, thymol test, increases the activity of catalase and the overall anti-radical activity, in the liver homogenate and blood serum inhibits the production of MDA;

2. equally reduces the content of DC, MDA, APL in the blood serum, and DC, AFL, OL in the liver homogenate.

Example 2

10.0 g of raw material, crushed to a particle size of 1-2 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The content of PL is 2.10% and FCC is 1.30%.

Example 3

10.0 g of raw materials, crushed to a particle size of 4-6 mm, consisting of the leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heat at 80 ° C for 1 hour, then cool to room temperature, separate obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.50% and FCC is 1.40%.

Example 4

10.0 g of raw materials, crushed to a particle size of 6-8 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.30% and FCC is 1.20%.

Example 5

10.0 g of raw material, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, is filled with 200 ml of 70% ethanol and heated at 20 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 0.50% and FCC is 0.50%.

Example 6

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at 40 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 1.45% and the FCC is 0.91%.

Example 7

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 60 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The content of PL is 2.10% and FCC is 1.20%.

Example 8

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 70 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.63% and FCC is 1.51%.

Example 9

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 75 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.80% and the FCC is 1.62%.

Example 10

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 85 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.90% and FCC is 1.58%.

Example 11

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 30 minutes, then cooled to room temperature, separated the obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 1.20% and FCC is 0.70%.

Example 12

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 45 minutes, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.15% and FCC is 1.30%.

Example 13

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 1.5 hours, then cooled to room temperature , the resulting extract is separated from the feed by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.90% and FCC is 1.65%.

Example 14

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 100 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.55% and FCC is 1.45%.

Example 15

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 150 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.70% and FCC is 1.50%.

Example 16

10.0 g of crushed raw material to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 250 ml of 70% ethanol and heat at 80 ° C for 1 hour, then cool to room temperature, separate the resulting extract from raw materials by straining The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.95% and FCC is 1.65%.

Example 17

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 300 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated twice more similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.95% and FCC is 1.66%.

Example 18

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration and the solvent is removed in vacuo.

The PL content is 1.95% and FCC is 0.94%.

Example 19

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. Separate extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated once again similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.52% and FCC is 1.44%.

Example 20

10.0 g of raw materials, crushed to a particle size of 2-4 mm, consisting of leaves, stems and flowers of a rough cornflower, pour 200 ml of 70% ethanol and heated at a temperature of 80 ° C for 1 hour, then cooled to room temperature, separated obtained extract from raw materials by filtering. The separated extract is purified from mechanical impurities by filtration.

Next, the extraction of the processed raw materials is repeated three more times similarly to the extraction described above. The resulting extracts were combined and the solvent was removed in vacuo.

The PL content is 2.90% and FCC is 1.65%.

SUBSTANTIATION OF THE MODE

In the process of searching for optimal conditions for obtaining an extract from plant material, consisting of crushed leaves, flowers and stems of rough cornflower, the influence of the following factors on the hepatoprotective and antioxidant activity and content of flavonoids and phenolcarboxylic acids was studied: the degree of grinding of the raw material, the ratio of raw material to extractant, and the temperature of extraction , extraction time and extraction ratio.

Studies of the antioxidant activity of aqueous, 40%, 70% and 96% ethanol extracts from the aerial part of the cornflower rough with a voltammetric method in vitro showed that all the extracts of the cornflower rough are more or less possess antioxidant properties, which are most pronounced in the extract obtained by 70% ethanol (table 2). Based on these data, 70% ethanol was used as an extractant.

It was found that extraction should be carried out with 70% ethanol with a ratio of raw materials-extractant 1: 15-1: 20, the degree of grinding of raw materials 2-4 mm, temperature 75-80 ° C for 1.0-1.5 hours and multiplicity extraction 3, followed by removal of the extractant in vacuo to obtain a dry extract.

When the ratio of raw materials-extractant 1:10 there is a low content of PL and PCA in the extraction, and extraction under these conditions is impractical. At ratios of 1:15, 1:20 and 1:25, the yield of the target product increases markedly. A further increase in the ratio (1:30) does not lead to a significant increase in the yield of PL and PCA (table 8), therefore, we recommend that extraction be carried out at a ratio of raw material-extractant 1: 15-1: 20.

With an increase in the extraction temperature to 75-80 ° С, the yields of PL and PCA significantly increase (by 1-1.5%) in comparison with the yields at an extraction temperature of 20-70 ° С. When heated to 85 ° C, the PL and FCC yields remain virtually unchanged, therefore, we recommend that extraction be carried out at a temperature not exceeding 75-80 ° C (table 6).

When extracting the aerial part of the rough cornflower for 30 minutes, the yield of PL and FCC significantly decreases (by 40%) compared with the duration of extraction for 1 hour, therefore, we recommend that the extraction be carried out for 1.0-1.5 hours. Extraction for 2 hours does not lead to an increase in the yield of PL and FCC (table 7).

When conducting a single extraction, the yield of PL and FCC significantly decreases. When carrying out 3- and 4-fold extraction, the yield of PL and FCC increases (table 9), but the use of four-fold extraction is not rational in terms of time spent to obtain the target product and a larger volume of the obtained liquid extract, requiring significant energy consumption for its evaporation. Therefore, we recommend a 3-time extraction.

The extracts obtained from the aerial part of the cornflower, rough at 70% ethanol in doses of 100 and 200 mg / kg, have hepatoprotective and antioxidant properties and helps normalize free radical oxidation of lipids. Extracts of rough cornflower in the indicated doses statistically significantly reduce the activity in the blood serum of ALT, ACT, as well as the content of total and conjugated bilirubin (table 2). These effects were obtained by increasing the total antioxidant activity, increasing the activity of catalase, and reducing the products of lipid peroxidation (diene conjugates and malondialdehyde) in rats with toxic liver damage with carbon tetrachloride (table 3). Therapy of rough cornflower extracts was accompanied by a regression of biochemical parameters.

Rough cornflower extract also has a positive effect on lipid metabolism. Extracts in all doses restore the level of OL and AFL in blood serum and liver tissue (table 4).

application

Table 1 - Antiradical activity of extracts of cornflower rough

Table 2 - The effect of the extract of cornflower rough (VS) on 70% ethanol on the biochemical parameters in the blood serum of toxic damage to the liver of rats with carbon tetrachloride (M ± m)

Note:

1 - p≤0.05 significant differences in relation to counter 1;

2 - p≤0,05 significant differences in relation to control 2,

3 - p≤0.05 significant differences in relation to hepatitis 1;

4 - p≤0.05 significant differences in relation to hepatitis 2.

Table 3 - The effect of the extract of rough cornflower (VS) on 70% ethanol on the processes of lipid peroxidation in blood serum and liver with toxic damage to the liver of rats with carbon tetrachloride (M ± m)

Note:

1 - p≤0.05 significant differences with respect to control 1;

2 - p≤0.05 significant differences with respect to control 2;

3 - p≤0.05 significant differences in relation to hepatitis 1;

4 - p≤0.05 significant differences in relation to hepatitis 2.

Table 4 - The effect of the extract of rough cornflower (VS) on 70% ethanol on the processes of lipid peroxidation in blood serum and liver with toxic damage to the liver of rats with carbon tetrachloride (M ± m)

Note:

1 - p≤0.05 significant differences with respect to control 1;

2 - p≤0.05 significant differences with respect to control 2;

3 - p≤0.05 significant differences in relation to hepatitis 1 and hepatitis 2;

4 - p≤0.05 significant differences in relation to hepatitis 2.

Table 5 - Effect of the degree of grinding of raw materials on the content of PL and FCC in the extract (in terms of air-dry raw materials)

Note: extraction temperature - 80 ° C; extraction time - 1 hour; extraction ratio - 3; the ratio of raw material-extagent is 1:20.

Table 6 - Effect of temperature on the content of PL and FCC in the extract (in terms of air-dry raw materials)

Note: the degree of grinding of raw materials - 2-4 mm; extraction time - 1 hour; extraction ratio - 3; the ratio of raw material-extagent -1: 20.

Table 7 - Effect of extraction time on the content of PL and FCC in the extract (in terms of air-dry raw materials)

Note: the degree of grinding of raw materials - 2-4 mm; extraction temperature - 80 ° C; extraction ratio - 3; the ratio of raw material-extagent is 1:20.

Table 8 - the Effect of the ratio of raw materials-extractant on the content of PL and FKK in the extract (in terms of air-dry raw materials)

Note: the degree of grinding of raw materials - 2-4 mm; extraction temperature - 80 ° C; extraction time - 1 hour; the extraction ratio is 3.

Table 9 - The effect of the extraction ratio on the content of PL and FCC in the extract (in terms of air-dry raw materials)

Note: extraction temperature - 80 ° C; extraction time - 1 hour; extraction ratio - 3; the ratio of raw material extractant is 1:20.

Table 1 Reaction time, min Water extract Extract on 40% ethanol 70% ethanol extract 95% ethanol extract Dihydroquerce-
Tina 5 69.53 ± 3.27 67.8 ± 1.86 58.55 ± 0.50 76.01 ± 2.20 9.15 ± 7.25 fifteen 67.68 ± 1.91 63.41 ± 1.65 23.40 ± 1.12 72.07 ± 2.70 6.86 ± 5.05 25 65.47 ± 0.89 60.84 ± 1.21 18.47 ± 0.57 69.70 ± 3.53 4.41 ± 1.70 35 63.87 ± 0.74 59.15 ± 1.25 14.53 ± 0.32 67.80 ± 3.45 - 45 62.96 ± 0.87 57.54 ± 1.30 10.82 ± 0.88 66.45 ± 3.80 - 55 61.59 ± 1.36 56.19 ± 1.23 7.89 ± 0.54 63.99 ± 4.99 - 60 60.78 ± 2.42 55.62 ± 1.43 6.53 ± 0.88 62.98 ± 4.37 -

table 2 Experimental
ny groups (n = 6-10) The studied indicators ALT ACT Bilirubin, μmol / L Alkaline phosphatase, μmol / (min · l) Thymol test, SH / unit E / L common conjug. Control 1 (water purification.) 10.27 ± 1.35 9.25 ± 1.43 39.88 ± 9.36 4.05 ± 3.47 136.13 ± 25.01 3.27 ± 1.07 Control 2 (vegetable oil + starch. Mucus) 10.01 ± 1.42 11.96 ± 0.36 32.65 ± 8.37 7.54 ± 1.28 116.58 ± 41.84 3.25 ± 2.02 Hepatitis 1 (CCI ₄ was administered 4 days) 22.08 ± 1.40 ¹ 15.49 ± 0.78 ¹ 77.33 ± 16.74 ¹ 18.31 ± 5.16 ¹ 194.93 ± 58.08 ¹ 13.27 ± 6.13 ¹ Hepatitis 2 (CCI ₄ was administered 4 days + starch. Mucus 6 days of administration) 13.75 ± 3.56 ² 13.49 ± 1.95 ² 50.75 ± 0.01 ² 14.02 ± 7.38 ² 117.50 ± 38.57 ² 3.25 ± 2.02 ² Karsil, 100 mg / kg 12.09 ± 1.02 ³ 13.05 ± 0.63 ³ 26.10 ± 3.97 ^3.4 2.93 ± 0.33 ^3.4 155.5 ± 13.28 ^3.4 3,50 ± 0,58 ³ 100 mg / kg VSH extract 10.39 ± 0.82 ³ 12.68 ± 0.51 ^3.4 40.07 ± 5.40 ^3.4 3.31 ± 1.10 ^3.4 134.17 ± 18.76 ³ 2.24 ± 0.53 ^3.4 200 mg / kg VS extract 11.37 ± 0.98 ³ 11.38 ± 1.99 ^3.4 35.54 ± 10.35 ^3.4 2.90 ± 0.48 ^3.4 116.76 ± 36.50 ³ 0.96 ± 0.39 ^3.4 400 mg / kg VS extract 10.98 ± 1.53 ³ 10.64 ± 2.65 ^3.4 38.67 ± 9.65 ^3.4 7.70 ± 4.35 ^3.4 92.5 ± 24.19 ^3.4 4.07 ± 1.58 ³

Table 3 Experimental groups (n = 6-10) The studied indicators MDA DK Catalase, μmol / sec-ml AOA, Radicalsvyaz. act% Liver, mmol / g Serum, mcmol / ml Liver, cu / g Serum, cu / ml Control 1 (water purification.) 16.6 ± 9.80 0.89 ± 0.40 89.54 ± 4.63 7.82 ± 0.47 154.84 ± 31.16 12.03 ± 3.09 Control 2 (vegetable oil + starch. Mucus) 15.92 ± 3.19 1.47 ± 0.55 84.08 ± 1.85 8.32 ± 0.39 161.9 ± 11.90 14.72 ± 1.27 Hepatitis 1 (CCI ₄ ) 24.76 ± 5.87 ¹ 2.05 ± 0.79 ¹ 108.75 ± 3.23 ¹ 9.80 ± 0.41 ¹ 84.59 ± 6.27 ¹ 4.1 ± 0.99 ¹ Hepatitis 2 (CCI ₄ + collapse. Mucus) 24.69 ± 10.13 ² 2.80 ± 0.88 ² 94.42 ± 3.55 ² 8.38 ± 0.57 ² 139.3 ± 5.67 ² 10.83 ± 2.65 ² Karsil, 100 mg / kg 28.08 ± 7.35 1.27 ± 0.13 ^3.4 85.43 ± 3.75 ^3.4 8.02 ± 0.87 ³ 145.93 ± 30.01 ³ 15.43 ± 0.90 ^3.4 100 mg / kg VSH extract 8.1 ± 2.66 ^3.4 0.86 ± 0.31 ^3.4 85.75 ± 1.36 ^3.4 8.63 ± 0.38 ³ 153.76 ± 12.00 ^3.4 22.68 ± 8.77 ^3.4 200 mg / kg VS extract 19.59 ± 8.06 ^3.4 1.59 ± 0.58 ^3.4 85.00 ± 1.36 ^3.4 8.44 ± 0.31 ³ 153.76 ± 12.00 ^3.4 22.90 ± 6.00 ^3.4 400 mg / kg VS extract 22.91 ± 2.70 2.41 ± 0.39 107.25 ± 6.59 9.65 ± 0.50 145.95 ± 22.02 ³ 22.47 ± 5.18 ^3.4

Table 4 Experimental groups (n = 6-10) The studied indicators GENERAL LIPIDS PHOSPHOLIPIDS Liver, mg / g Serum g / l Liver, mg / g Serum g / l Control 1 (water pur.) 34.96 ± 6.14 2.93 ± 0.77 11.89 ± 1.35 0.39 ± 0.16 Control 2 (vegetable oil + starch. Mucus) 30.87 ± 12.97 3.26 ± 0.77 11.74 ± 0.72 0.41 ± 0.08 Hepatitis 1 (CCl ₄ ) 67.16 ± 10.69 ¹ 4.00 ± 0.10 ¹ 23.97 ± 3.54 ¹ 1.01 ± 0.03 ¹ Hepatitis 2 (CCl ₄ + collapse. Mucus) 48.06 ± 2.33 ² 3.60 ± 0.22 ² 18.67 ± 5.23 ² 0.80 ± 0.26 ² Karsil, 100 mg / kg 36.58 ± 1.56 ^3.4 2.38 ± 0.41 ^3.4 12.22 ± 1.17 ^3.4 0.59 ± 0.25 ^3.4 100 mg / kg VSH extract 43.48 ± 3.10 ³ 3.38 ± 0.55 ³ 12.61 ± 0.88 ^3.4 0.29 ± 0.05 ^3.4 200 mg / kg VS extract 32.88 ± 2.60 ^3.4 2.93 ± 0.21 ^3.4 11.15 ± 1.78 ^3.4 0.44 ± 0.08 ^3.4 400 mg / kg VS extract 43.63 ± 3.07 3.48 ± 0.22 ³ 11.86 ± 0.84 ^3.4 0.31 ± 0.03 ^3.4

Table 5 No. p / p The degree of grinding of raw materials, mm Content% FL, in terms of routine FCC, calculated on caffeic acid one 1-2 2.10 1.30 2. 2-4 2.85 1,60 3. 4-6 2.50 1.40 four. 6-8 2,30 1.20

Table 6 No. p / p Temperature ° C Content% FL, in terms of routine FCC, calculated on caffeic acid one. twenty 0.50 0.50 2. 40 1.65 0.91 3. 60 2.10 1.20 four. 70 2.63 1.51 5. 75 2.80 1,62 6. 80 2.85 1,60 7. 85 2.90 1,58

Table 7 No. p / p Extraction time, min Content% FL, in terms of routine FCC, calculated on caffeic acid one. thirty 1.20 0.70 2. 45 2.15 1.30 3. 60 2.85 1,60 four. 90 2.90 1.65

Table 8 No. p / p The ratio of raw material extractant Content% FL, in terms of routine FCC, calculated on caffeic acid one. 1:10 2,55 1.45 2. 1:15 2.70 1,50 3. 1:20 2.85 1,60 four. 1:25 2.95 1.65 5. 1:30 2.95 1.66

Table 9 No. p / p Extraction rate Content% FL, in terms of pa FCC, calculated on caffeic acid one. one 1.95 0.94 2. 2 2,52 1.44 3. 3 2.85 1,60 four. four 2.90 1.65

Sources of information taken into account

1. Industrial technology of drugs: a Textbook in 2 volumes. Volume 2 / V.I. Chueshov, M.Yu. Chernov, L.M. Khokhlova and others; Ed. Professor V.I. Chueshov. - Kharkov: publishing house MGK-Book; NFAAU, 2002 .-- S.158-159.

2. Guide to experimental (preclinical studies) of new pharmacological substances / Ed. member - correspondent. RAMS prof. R.U. Khabrieva. - 2 - ed., Revised. and add. - M .: Medicine, 2005 .-- 836 p.

3. Vladimirov Yu.A. Lipid peroxidation in biological membranes / Yu.A. Vladimir, A.I. Archov. - M .: Nauka, 1972.- 239 p.

4. Kamyshnikov B.C. Clinical diagnostic methods in biochemistry / V.S. Kamyshnikov. - M .: Nauka, 2002. -568 p.

5. Katikova O.Yu. The effect of herbal remedies on LPO indicators in acute toxic hepatitis / O.Yu. Katikova // Questions of medical chemistry. - 2001. - T. 47, No. 6. - S.593-598.

6. Cosman V.M. Quantitative extraction spectrophotometric determination of the total content of hydroxycinnamic acids in the presence of flavonoids in the extractives of some medicinal plants / V.M. Kosman, I.G. Zenkevich // Plant Resources. - 2001. - T.37, No. 4. - S. 123-129.

7. State Pharmacopoeia of the USSR: Issue 2. General methods of analysis. Medicinal plant material / Ministry of Health of the USSR. - 11th ed. - M .: Medicine, 1990. - 337 p.

Claims (2)

1. The use of the extract from the aerial part of the rough cornflower (Centaurea scabiosa L.) as a hepatoprotective agent with antioxidant action. 2. A method of producing a hepatoprotective agent with an antioxidant effect, characterized in that the aerial part of the rough cornflower (Centaurea scabiosa L.), crushed to a particle size of 2-4 mm, is extracted three times with 70% ethanol at a temperature of 75-80 ° C for 1 -1.5 hours, with a ratio of raw materials-extractant 1: 15-1: 20, followed by evaporation of the obtained extract to dryness.