SU1751669A1 - Method for determining toxicity of water and aqueous solutions containing active substances - Google Patents

Abstract

The use of environmentalists, the control of the toxicity of water and aqueous solutions. The essence of the invention: the determination of toxicity is carried out by exposing the test water and aqueous solutions to cells of high pH. leaflets and plant contents of light for 17–18 h with illumination of 50 ± 4 lx in the test liquid. Then the number of epistrophic chloroplasts is counted before and after 10 minutes of exposure to light. close to the maximum for this type of microscope The ratio of chloroplasts remaining in the epistrophic position to the initial level is an indication of the toxicity of water or aqueous solutions

Description

The invention relates to environmental issues, namely the control of the toxicity of water or aqueous solutions.

Methods are known for determining the toxicity of water and aqueous solutions by their effect on living organisms: animals (mammals, fish, invertebrates, protozoa), or plant cells. (one,

2).

The known method consists in the action of harmful environmental substances on plant cells (protoplasts) of Vlciafaba, Avenasativa immobilized in the matrix of Ca-alginate or Za-alyinate. The intensity of exposure to harmful substances is judged by changes in the biochemical and physiological properties of cells, namely, ethane is determined in 1 m.

gas sample by ethane-gas chromatography or by content of C1402 in mmol per mg chlorophyll per hour using a scintillation counter.

The disadvantages of this method are the complexity of the technology (the production or cultivation of protoplasts, the preparation of matrices, the immobilization of cells into matrices); the use of expensive and scarce chemicals (enzymes, labeled precursors, etc.); low sensitivity of the method.

The purpose of the invention is to increase the sensitivity of the method

The goal in the method of determining the toxicity of water and aqueous solutions containing biologically active substances, by the impact of the test

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a fight on plant cells with a subsequent assessment is achieved by effecting a part of the test sample being studied into the intercellular spaces of the leaves, then the plants are kept in the test liquid for 17-18 h at illumination 50 ± 4 lux, then the number of epistrophic chloroplasts before and after, 10-minute exposure to light and assess toxicity by increasing the number of epistrophic chloroplasts,

Example. For the study, leaflets of color (Zemna minor Z) are infiltrated with various water samples (examples 1 through 5) using a glass medical syringe and placed on a weak light (50 ± 4) lux for 17-18 hours.

One leaf is detached from the plants, which are placed in a drop of the test liquid on a microscope slide and at 400 times magnification in 10 cells of the upper side of the leaf, in one and the same field of view of the microscope, the number of chloroplasts that are positive phototaxis at the upper cell wall in an epistrophic position perpendicular to the rays of light, flat. The preparation doesn’t shift; in a microscope for 10 minutes they create a strong illumination of the object, which is close to the maximum (almost the intensity of the lamp of the illuminator increases to the limit).

After 10 minutes, the illumination of the object is reduced to a convenient limit and the number of epistrophic chloroplasts is again calculated in the cells.

In the control (tap or distilled water), the number of such chloroplasts usually approached 0, since most of the chloroplasts turned into a parastrophic position as a result of negative phototaxis, i.e. pressed against the side walls of the cells, turning edge with respect to the rays of light. The more water is polluted, the more is preserved in the epistrophic chloroplast cells. The percentage of epistrophic chloroplasts after a 10-minute exposure to strong light in relation to the initial level is an indicator of damage to the cells by contaminated with water or aquatic plants and, therefore, an indication of the toxicity of water or aqueous solutions. The criterion of the direct toxic effect on the plant kpetk should be considered as a significant difference of the average indicators (in 10 cells of the leaves of the river) 5 experiments (parallels) from the control.

Under the conditions of the norm (without the action of a toxicant), the cell chloroplasts have an individual chow phototaxis, which is reflected in the insignificant scatter of the amount of chloroplasts when they are counted after a short light

However, this indicator is always significantly less than when exposed to toxicants. When determining the toxicity of substances in aqueous solutions, the criterion of toxic effect is also the minimum

concentration from the radiated series, which still causes a significant inhibition of phototaxis compared with the control

The choice of 10 min exposure is related to the fact that during this time with a strong

When illuminated, chloroplasts of control samples will move to an epistrophic position. This is the optimal time for negative chloroplast phototaxis to be normal. Exceeding this time is not advisable.

Example 1: Determination of the toxicity of wastewater entering the treatment plant. The counting of epistrophic (located plasm) chloroplasts in 10 cells of lista rk immediately after a weak light during

17h gave the following numbers: 20, 15, 24, 20.18. 21, 17, 24, 25 19. On average per cell 20.3 chloroplast. After 10 minutes of strong light, counting epistrophic chloroplasts in 10 cells in the same field.

From the view of the microscope, he gave the following figures: 14, 15, 11, 13, 9, O, O, 4, 0. 22. On average, 8.8 chloroplasts per cell, which amounted to 33% as a percentage of the initial value (counting after a dim light) . Last digit and

served as an indicator of phototaxis intensity or degree of cell damage.

In the control (tap water) the following data was obtained after a weak light: 24, 13, 17, 28, 16, 16, 17, 18, 26, 23.

The average per cell is 19.8 chloroplast. After 10 minutes of strong light: 4, 6, O, O, O, O, O, O, 0. 0. On average, there is 1.0 chloroplast per cell, which is 5% of the initial value,

Data from 5 experiments to determine the toxicity of the inputs at the entrance to the treatment plant gave the following average figures (as a percentage of the initial number of epistrophic chloroplasts): 43, 36, 49, 48, 27.

Similar data for tap water: 5, 12, 2, 0, 3. M ± m (4 4 ± 2,3)%. Reliability of the difference of water before cleaning from tap water 001

Example 2. Data from 5 experiments with water after cleaning before dropping into the Neva River (also as a percentage of the initial amount of chloroplasts) - 18 16. 26, 12, 18. M ± ± 27 Reliability difference

from tap water, 001, reliability difference from water before cleaning, 01

PRI me R 3. Similar data from 4 experiments with a water fraction of activated sludge, after its two-fold dilution: 66%, 78%, 90%, 100%, M ± m-83.5 ± 10.7. Reliability of greater toxicity of the aqueous fraction of activated sludge (after dilution by 2 times) compared with wastewater before purification:, 01.

PRI me R 4. Data from 4 experiments with Amphotericin B solution (1 mg / L) as a percentage of the initial amount of chloroplasts: 42, 29, 81, 100. M ± 17.2. Reliability differs from tap water:, 02.

Example 5: Data on the effect of heavy metal salts on chloroplasts on phototaxis (data on minimum concentrations), CuSCM solution (10 M). Obtained in% of the initial amount of chloroplasts: control M ±, 1 ± 0.3; experience M ± m 7.9 ± 2,, 05.

Solution (), obtained: control M ±, 4 ± 0.5; experience M ±, 5 ± 6; , 001. The toxicity of pentachlorophenol to immobilized Vicla faba protoplasts was radiated by a known method (no determination of ethane release relative to the control). Under the action of 14 mM pentachlorophenol, ethane production was increased about 2 times, which was an indicator of the toxicity of substances. According to the invention, phenol (less toxic than its chlorine derivatives) using the concentration of 7 mM gave the following results: control: 2, 1, 1, 0, 2 (% inhibition of chloroplast phototaxis) M, 34 ± 0.2; experience 8, 47, 15. 53. 76

0 M, 8 ± 12; .01.

Thus, the proposed method is much more sensitive than the prototype method.

Claims (1)

Invention Formula 5 A method for determining the toxicity of water and aqueous solutions containing biologically active substances by affecting the test sample on plant cells, followed by an assessment that differs so that, in order to increase the sensitivity of the method, the effect is carried out by introducing part of the test sample into the intercellular spaces of the leaves. then the plants are kept in the test liquid for 17–18 h at an illumination of 50+ 4 lx, then the number of epistrophic chloroplasts is counted before and after 10 minutes of exposure to light and tsenivayut toxicity to increase the number of chloroplasts epi0 strofnyh