RU2280466C1 - Method for preparing cellular sap from microcarpous plume poppy (macleya) above-ground part - Google Patents

Abstract

FIELD: cosmetic industry, phytotherapy.

SUBSTANCE: invention relates to a method for preparing cellular sap from microcarpous plume poppy above-ground part that involves milling the raw, twice extraction with a liquid consisting of 0.1% aqueous solution of NaCl with pH = 1-4 heated to temperature 80-90°C in the ratio of raw to the extractant volume = 1:0.5. Before squeezing out the primary sap is infused for 60-120 min in the constant stirring followed by squeezing the primary sac, then before squeezing the secondary sac is infused for 30 min followed by squeezing the secondary sap followed by combining the primary and secondary sap, sap preserving, its settling and filtration. Method provides preparing cellular sap with the enhanced content of biologically active substances.

EFFECT: improved preparing method.

2 tbl, 3 ex

Description

The invention relates to the cosmetic industry, in particular to a method for producing cell juice from plant materials intended for use as a biologically active additive in the manufacture of cosmetic creams for face and body, hair shampoos, toothpastes and hygienic lipsticks.

A known method for producing sanguine retrin (a mixture of benzo [c] phenanthridine alkaloids sanguinarine and chelerythrin hydrosulfates), which consists in the fact that the aboveground part of the small-fruited Macleaya (Macleaya microcarpa) or heart-shaped Macleaya (Macleaya cordata) is crushed and extracted with a hydrophobic solvent in the presence of. Then, the hydrosulfates of the target alkaloids are precipitated, the mixture of alkaloids salts is purified by treating the filtered and alkalized aqueous solution with an organic solvent, followed by treating the organic phase with an aqueous solution of sulfuric acid, and the precipitate is separated by filtration and washing with acetone. Ethyl acetate or dichloroethane is used as the extractant in the solid-phase extraction step. The extraction is carried out by sequential processing of several portions of the raw material with one volume of extractant according to the principle of countercurrent flow with the formation of extracts enriched with alkaloids. During purification, an aqueous solution of ammonia or potash is used as the alkaline agent in the extraction stage, and ethyl acetate or toluene is used as the extractant. Extraction purification is carried out at a temperature of 50-60 ° C at a rate that prevents the formation of a precipitate of the target alkaloids. The resulting extract is filtered, washed with water.

A known method of producing sanguirythrin, which consists in the extraction of crushed Macleia grass treated with ammonium hydrate, dichloroethane, filtration, subsequent processing of the extract with diluted sulfuric acid, separation of the precipitated hydrosulfates of alkaloids and their recrystallization from acidified alcohol (RF patent 571268, A 61 K 35/78) .

A known method of producing sanguirytrin from makleia grass of small-fruited and heart-shaped by multi-stage extraction of raw materials with chloroform in an alkaline medium with the gradual removal of a third of the volume of the extract from the extractor for the subsequent stage of treatment with sulfuric acid. The spent organic phase is returned to the extractor, and the aqueous-acid phase is returned to the processing of a new portion of the extract (RF application No. 93057354/14, A 61 K 35/78).

The disadvantages of the methods (RF patent 2141837, A 61 K 35/78, 31/435, RF patent 571268, A 61 K 35/78, RF application No. 93057354/14, A 61 K 35/78) is that these processes They are characterized by high labor intensity, energy intensity and do not meet environmental requirements, since they imply the use of toxic extractants (dichloroethane, toluene, chloroform). In addition, these processes are aimed at obtaining a pharmaceutical drug sanguirytrin, which excludes the extraction of other biologically active substances contained in the grass of makleia.

A known method of obtaining juice from freshly collected nettle leaves, which consists in the fact that the raw material is crushed, it is mechanically pressed. Then the resulting juice is heated to inactivate the enzymes. Preservation of juice is carried out using ethyl alcohol in an amount of 15% and chloroethane. Then the juice is defended and filtered (Actual problems of creating new drugs of natural origin. Congress materials. - St. Petersburg: Design, 2000. S.95-97).

The disadvantage of this method is that such a process does not allow to achieve maximum extraction of biologically active substances, since there is no stage of preliminary processing of raw materials before mechanical extraction. Thus, cell membranes are not destroyed, and the sorbed and chemically bonded components of the solid phase remain unchanged. Therefore, it is not possible to achieve a deep extraction of biologically active substances.

Closest to the claimed is a method of obtaining cell juice from technical greens, which consists in the fact that the crushed technical greens are treated with hot steam, mechanical extraction of the raw materials is carried out to obtain cell juice. The resulting juice is separated from depleted raw materials, canned with ethyl alcohol in an amount of 20%, sedimented and filtered (Tomchuk R.I., Tomchuk G.N. Woody greens and its use in the national economy. - M.: Forest industry, 1973. 360 p. .).

The disadvantage of this method is that in this case, raw steaming is used, which leads to a significant destruction of biologically active substances such as vitamins, organic acids, pigments.

The objective is to create a method for producing cell juice from plant materials, namely, from the aerial part of the small-fruited mackley, containing a complex of biologically active substances of this plant, such as sanguinarine hydrosulfate, chelerythrin hydrosulfate, ascorbic acid (vitamin C), tannins.

The technical result of the invention is the deep extraction of a complex of biologically active components from plant materials, namely from the aerial part of the small-fruited mackley.

The technical result is achieved by the fact that in the method of producing cell juice from plant materials, including grinding the raw materials, processing with an extractant, mechanical extraction of the raw materials, separating the cell juice from the depleted raw materials, preserving the juice, settling and filtering it, the aerial part of the small-fruited macklea is extracted with twice electroactivated liquid, which is an electroactivated 0.1% aqueous NaCl solution with a pH of 1-4, heated to 80-90 ° C with a ratio of the mass of raw materials to the volume of extractant 1: 0.5, before pressing the primary juice is infused for 60-120 minutes with constant stirring, before pressing the secondary juice is insisted for 30 minutes, combine the primary and secondary juice.

The operation of preparing raw materials using electroactivated liquid is key to ensuring the deep and intensive extraction of biologically active substances contained in small-fruited mackley. When insisting crushed raw materials in an electroactivated liquid with a pH of 1-4, hydrolytic cleavage of the cellulose membranes surrounding plant cells occurs. Thus, the cell is partially destroyed, the state of sorbed and chemically bonded compounds, as well as their position in space, change, which contributes to the intensification of the process of mass transfer from intracellular to extracellular space. In addition, the alkaloids sanguinarine and cheleritrin (the main active ingredients of macklea small-fruited) are supported in a biologically active form - in the form of an acid salt, thereby increasing the biological activity of the juice.

Subsequent mechanical extraction of raw materials prepared using electroactivated liquid with a pH of 1-4, allows you to get the cellulose juice of small-fruited, containing a complex of biologically active substances of the plant.

The possibility of implementing the method is shown in examples of specific performance.

Example 1

To obtain cell juice, the aerial part of the small-fruited mackley, an electroactivated liquid with a pH of 2.5, heated to 85 ° C, was used.

The aerial part of the small-fruited mackley is a mixture of pieces of stems, leaves and buds. Pieces of stems up to 15 cm long and 1.5 cm in diameter, cylindrical in shape, externally yellowish-gray to brownish-gray in color, sometimes with a wax coating; a yellowish-brown crustal part and a white friable core are visible in a cross section. Pieces of leaves of various shapes up to 10 cm in size from brownish-green to brownish-yellow or grayish-green. Buds up to 0.7 cm long, cylindrical yellowish-brown in color with a faint odor.

Physico-chemical parameters of small-fruited mackley are shown in table 1.

Table 1
Physico-chemical indicators of small-fruited makleya Name of indicator Characteristic and norm Mass fraction of moisture,%, not less than 70 Mass fraction of organic impurity,%, no more one Mass fraction of mineral impurity,%, no more one Mass fraction of sanguirythrin (a mixture of sanguinarine and helerythrin hydrosulfates),% on absolutely dry mass, not less than 0.6

Small-fruited Macleia is rich in alkaloids, which have a pronounced antimicrobial activity. The main alkaloids of the aerial part of the plant are chelerythrin and sanguinarine (up to 0.8% on absolutely dry weight). In addition to alkaloids, makleia grass contains such biologically active substances as organic acids, phenolic compounds, vitamins, and pigments.

An electroactivated liquid with a pH of 2.5 was obtained by electrolysis of a 0.1% aqueous NaCl solution in a laboratory installation for electroactivation of liquids. The productivity of the installation is 1.2 l / h, the power consumption is not more than 5 W, the power is supplied from the 220 V network.

An aerial part of small-fruited mackley with a mass fraction of moisture of 75% in an amount of 100 g was loaded into a 500 ml container with a grinder. An electroactivated liquid with a pH of 2.5 in an amount of 50 ml was poured into the container with stirring. The resulting mass was kept for 90 minutes with constant stirring. After the specified time, the mass was fed to the laboratory hydraulic press IP 6010-100-1, where it was mechanically pressed. The obtained cell juice was sent to a cell juice collector, and the squeezed raw materials were again treated with an electroactivated liquid with a pH of 2.5, aged for 30 minutes with stirring, and fed to a press. The resulting juice also entered the cell juice collection. Then the juice was canned, followed by the stage of sedimentation and filtration.

The mass fraction of sanguirythrin (a mixture of sanguinarine and helerythrin hydrosulfates), the mass fraction of ascorbic acid, and the mass fraction of tannins are accepted as an indicator of the quality of the obtained product.

The quantification of sanguirytrin in the cell sap was carried out by chromatospectrophotometric method (FS 42-2666-89. Macleia grass). Analysis progress: about 5 g (accurately weighed) of the juice is placed in a 250 ml conical flask with a ground stopper, 5 ml of 25% ammonia are added, mix thoroughly, close the stopper and leave at room temperature for 30 minutes. Then, 100 ml of chloroform was added to the flask, stirred, closed with a stopper and left for 15 hours. After this period, the contents of the flask are stirred for 10-15 minutes, left standing for 10 minutes, 50 ml of the extract are taken with a pipette, transferred to a 100 ml round bottom flask and the solvent is completely distilled off. The dry residue is dissolved in 5 ml of chloroform; 0.14 ml of the resulting solution was micropipetted onto the start line of the plate. The plate is dried in air for 20 minutes, placed in a chromatographic chamber with a mixture of benzene-chloroform-methyl alcohol-25% ammonia solution (100: 80: 1: 19), which is pre-saturated with solvent vapors for 1 h at a temperature of 20-25 ° С . After the solvent front reaches the end of the plate, it is removed and dried for 60 minutes in air. Zones containing sanguirythrin (orange-yellow) are scraped off the plate, quantitatively transferred to 50 ml flasks, 10 ml of acidified methyl alcohol are added, stirred for 5 minutes, then poured into centrifuge tubes and centrifuged for 10 minutes at 6000 rpm. min

The optical density of the solution is measured on a spectrophotometer in a cuvette with a layer thickness of 10 mm at 452 nm. As a comparison solution, acidified methyl alcohol is used.

At the same time, the optical density of the solution of the working standard sample (SEM) of sanguirytrin is measured.

Mass fraction of sanguirythrin X,%, calculated by the formula:

where: D ₁ is the optical density of the test solution;

D ₀ - the optical density of the solution of the SAR of sanguine;

m ₀ is the mass of sanguirytrin taken for the preparation of a solution of RNO, g,

m ₁ - weight of juice, g.

Quantitative determination of ascorbic acid was carried out by titration with Tilmans paint (Methods of biochemical study of plants / Edited by A.I. Ermakov. - L., 1987, 430 s). The course of analysis. A portion of cell juice (10 g) is dissolved in a 100 ml ³ volumetric flask with a 1% oxalic acid solution. The contents of the flask were adjusted to the mark with a solution of oxalic acid, covered with a stopper, shaken vigorously and left to stand for about 5 minutes. Then take 5 cm ^{3 of the} colored extract in test tubes (diameter 20-25 mm, length 150 mm) and chemically pure chloroform is added thereto with a graduated cylinder of 5 cm ³ . Then the extract is titrated from the microburette 0.001 N. Tillmans paint solution (2,6-dichlorophenolindophenol) with gentle stirring, tilting the tube covered with a stopper. When the first pink color appears in the chloroform layer, titration is considered complete. At the same time, titration of control samples is carried out with the difference that a 1% solution of oxalic acid is added to the tubes. 1 cm ³ of a paint solution corresponds to 0.088 mg of ascorbic acid.

Quantitative determination of tannins was carried out according to a standard method (State Pharmacopoeia of the USSR, X edition). The course of analysis. 2 g (accurately weighed) of the cell juice is dissolved in a 250 ml ³ volumetric flask with water. The contents of the flask are adjusted to the mark with water. 25 cm ^{3 of the} obtained liquid is placed in a 1 L conical flask, 750 cm ^{3 of} water, 25 cm ³ of indigosulfonic acid solution are added and titrated with constant stirring of 0.1 N potassium permanganate solution until golden yellow. 1 cm ³ 0.1 N. a solution of potassium permanganate corresponds to 0.004157 g of tannins in terms of tannin. In parallel, a control experiment is carried out, titrating 25 cm ^{3 of} indigosulfonic acid in 750 cm ^{3 of} water.

Examples 2 and 3.

In examples 2 and 3, the process of obtaining cellulose mackleia small-fruited juice was carried out according to the scheme described in example 1. The conditions for the process of preparing cell juice and the results of quantitative analysis of the obtained product are presented in table 2.

table 2
The conditions for the process of obtaining cell juice from the aerial part of macklea small-fruited by the present method Name of indicator Example 1 Example 2 Example 3 pH of electroactive liquid 2,5 one four Temperature of electroactivated liquid, ° С 85 90 80 The ratio of the mass of raw materials to the volume of extractant 1: 0.5 1: 0.5 1: 0.5 The duration of the process of infusion of the feedstock, min 90 120 60 The duration of the process of insisting depleted raw materials, min thirty thirty thirty Mass fraction of sanguirythrin,% 0.31 0.29 0.26 Mass fraction of tannins (in terms of tannin), mg / 100 g 92.34 75.32 69.28 Mass fraction of ascorbic acid, mg / 100 g 0.350 0.408 0.268

Claims (1)

A method of obtaining cell juice from plant materials, including grinding the raw materials, processing with an extractant, mechanical extraction of the raw materials, separating the cell juice from the depleted raw materials, preserving the juice, settling and filtering, characterized in that the aerial part of the small-fruited macklea is extracted twice with an electroactivated liquid, which is an electroactivated A 0.1% aqueous solution of NaCl with pH = 1-4, heated to 80-90 ° C with a ratio of the mass of raw materials to the volume of extractant 1: 0.5, insist before pressing the primary juice for 60-120 minutes with constant stirring before the secondary juice squeezer infused for 30 minutes, combined primary and secondary juice.