Neutral solvent for dissolving xanthine in soil experiment
By using neutral solvents composed of ammonia water, ethanol, hydrochloric acid and deionized water, these solvents are added in sequence to dissolve xanthine, which solves the problems of unstable and pH changes in the prior art, and achieves a clear and stable xanthine solution, which is suitable for soil experiments.
Patent Information
- Application Number
- CN202411830455.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-12
- Publication Date
- 2025-03-28
AI Technical Summary
The xanthine solution used in soil experiments in the prior art is difficult to completely dissolve xanthine, resulting in particle settlement and affecting the test effect. Commonly used solvents such as ammonia and hydrogen peroxide will change the soil pH and affect the experimental results.
A neutral solvent composed of ammonia water, ethanol, hydrochloric acid and deionized water in a specific volume ratio is used to dissolve xanthine by adding these solvents in sequence to ensure the stability and neutral properties of the solution.
The complete dissolution of xanthine is achieved, the resulting solution is clear and stable, without particulate matter, and will not change the soil pH during use, meeting the needs of soil experiments.
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Figure CN119715995A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of drug preparation, in particular to a neutral solvent used for dissolving xanthine in soil experiments. Background Art
[0002] Xanthine, a white to light yellow crystalline powder with a molecular formula of C5H4N4O2, exists in organisms mainly as a component of energy storage and bioactive molecules, and mainly exists in the process of nucleic acid catabolism. Xanthine itself has some biological activity, can participate in the signal transduction process in cells as a signal transduction molecule, and can also be decomposed into ATP to provide energy for cells. Since xanthine solutions can promote the growth of beneficial microorganisms in the soil and thus promote plant growth, xanthine solutions can also increase plant salt tolerance. Therefore, research on xanthine has moved from the laboratory to soil experiments.
[0003] When xanthine is used to carry out soil experiment, it is often necessary to add it into soil in the form of xanthine solution. Xanthine is soluble in sodium hydroxide solution, ammonia and acidic solution, slightly soluble in water and ethanol, and insoluble in organic solvent. In the prior art, xanthine is dissolved in water, ethanol or Tween-80 to be prepared into suspension for soil experiment, but when the suspension form is used for soil experiment, undissolved xanthine particles will gradually settle on the soil surface or shallow layer, and the contact with soil particles is relatively confined around the particles, affecting the test effect, while the solution form can make xanthine uniformly distributed in soil pores and particle surface, which is conducive to exerting its influence on various biochemical processes in soil. In addition, there is also a solvent such as ammonia, hydrogen peroxide and part of the acidic solution to dissolve xanthine, but when ammonia and hydrogen peroxide are used as solvents, the obtained xanthine solution is alkaline, and when it is used for soil experiment, it will destroy soil pH, thereby changing soil physicochemical properties. At the same time, the xanthine solution obtained by dissolving xanthine in an acidic solution is acidic. When it is used for soil tests, it will not only destroy the soil pH, but also be accompanied by insufficient dissolution, causing xanthine to precipitate in the form of particles, thereby affecting the soil experimental results.
[0004] Therefore, seeking a solvent that can be used to dissolve xanthine, which can not only ensure that it can completely dissolve xanthine to form a clear, stable solution without obvious particulate matter, but also ensure that the xanthine solution prepared by the above solvent does not affect the pH of the soil when used for soil testing, is a major problem that needs to be solved at present. Summary of the invention
[0005] In view of the above-mentioned prior art, the purpose of the present invention is to provide a neutral solvent for dissolving xanthine in soil experiments. The present invention adopts ammonia water, ethanol, hydrochloric acid and deionized water to form a neutral solvent for dissolving xanthine. The above-mentioned neutral solvent can achieve effective dissolution of xanthine, ensure that the obtained xanthine solution is clear and stable, free of particulate matter, and will not clog the needle when drawn with a syringe. At the same time, the neutral solvent of the present invention is safe and can meet the needs of soil experiments. Specifically, when the obtained xanthine solution is used in soil experiments, it will not cause pH changes in the soil, and the xanthine solution obtained by continuously applying it to the soil for 7 days will not cause changes in the physical and chemical properties of the soil.
[0006] To achieve the above object, the present invention adopts the following technical solution:
[0007] The first aspect of the present invention provides a neutral solvent for dissolving xanthine, wherein the neutral solvent is composed of ammonia water, ethanol, hydrochloric acid and deionized water in a volume ratio of (12-13): (2-3): (4-6): (78-82).
[0008] Preferably, the mass fraction of the hydrochloric acid is 15-25%, the mass fraction of the ammonia water is 15-25%, and the mass fraction of the ethanol is 15-25%.
[0009] The second aspect of the present invention provides the use of the above neutral solvent in preparing a xanthine solution.
[0010] The third aspect of the present invention provides a xanthine solution prepared by the following method:
[0011] Add ammonia water, ethanol, hydrochloric acid and deionized water to xanthine powder in sequence and mix well to obtain xanthine solution.
[0012] Preferably, the solid-liquid ratio of xanthine powder to ammonia water is (1-2) g: (12-13) L.
[0013] Preferably, the added amount ratio of xanthine powder, ammonia water, ethanol, hydrochloric acid and deionized water is (1-2) g: (12-13) L: (2-3) L: (4-6) L: (78-82) L.
[0014] The fourth aspect of the present invention provides the use of the above xanthine solution in maintaining soil stability.
[0015] Beneficial effects of the present invention:
[0016] The present invention adopts ammonia water, ethanol, hydrochloric acid and deionized water to form a neutral solvent for dissolving xanthine. The above neutral solvent can achieve effective dissolution of xanthine, ensure that the obtained xanthine solution is clear and stable, has no particulate matter, and does not clog the needle when the syringe is used for extraction. At the same time, the neutral solvent of the present invention is safe and can meet the needs of soil experiments. Specifically, when the obtained xanthine solution is used for soil experiments, it will not cause pH changes in the soil, and the xanthine solution obtained by continuously applying it to the soil for 7 days will not cause changes in the physical and chemical properties of the soil.
[0017] The present invention adopts the method of sequentially adding ammonia water, ethanol, hydrochloric acid and deionized water to xanthine powder, and the order of adding each component in the neutral solvent is crucial for obtaining a stable and effective xanthine solution. Specifically, since xanthine is soluble in ammonia water and slightly soluble in ethanol, ammonia water is first added to the xanthine powder to preliminarily dissolve the xanthine powder, and then ethanol is added as a co-solvent, and hydrochloric acid is used as a pH regulator. When a small amount of hydrochloric acid is added, ammonium chloride is generated, resulting in a reduction in the amount of ammonia water in the reagent, but does not cause xanthine to precipitate, and finally deionized water is added to adjust to the final concentration and ensure the stability of the solution. This configuration order ensures that the xanthine powder is fully dissolved and maintains the clarity of the solution, thereby ensuring the safety and effectiveness of the test. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] Figure 1 : Comparison of the dissolution effects of xanthine at different stages in Example 1, wherein a is xanthine powder, b is the first solution, c is the second solution, d is the third solution, e is the xanthine solution, and f is the xanthine solution placed at room temperature for 1 hour;
[0019] Figure 2 : The dissolution effect diagram of xanthine in neutral solution in Example 2;
[0020] Figure 3 : The dissolution effect diagram of xanthine in deionized water in Comparative Example 1;
[0021] Figure 4 : Comparative Example 2, the dissolution effect diagram of xanthine in ethanol;
[0022] Figure 5 : Comparative Example 3, the dissolution effect diagram of xanthine in hydrochloric acid;
[0023] Figure 6 : Comparative Example 4, the dissolution effect diagram of xanthine in ammonia water;
[0024] Figure 7 : The effect of the concentration of xanthine solution on the germination rate of soybean seeds in Experimental Example 1;
[0025] Figure 8: The growth-promoting effect of xanthine solutions of different concentrations on soybean in Experimental Example 1;
[0026] Fig. 9 : Graph showing the effect of xanthine solution on the pH of different soils in Experimental Example 2. DETAILED DESCRIPTION
[0027] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.
[0028] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.
[0029] The experimental materials used in the embodiments of the present invention are all conventional experimental materials in the art and can be purchased through commercial channels.
[0030] Example 1: Preparation of xanthine solution
[0031] The neutral solution is composed of 20% ammonia water, 20% ethanol, 20% hydrochloric acid and deionized water in a volume ratio of 12.5:2.5:5:80.
[0032] Place 1 μg of xanthine powder in an EP tube, add 12.5 μL of 20% ammonia water to the xanthine solution, mix well to obtain a first solution; add 2.5 μL of 20% ethanol to the first solution, mix well to obtain a second solution; add 5 μL of 20% hydrochloric acid to the second solution, mix well to obtain a third solution; add 800 μL of deionized water to the third solution, mix well to obtain a xanthine solution with a concentration of 1 μg / mL. In the preparation of the xanthine solution, flick the bottom of the tube after adding the solvent each time to fully dissolve the xanthine powder.
[0033] The dissolution effects of the xanthine powder, the first solution, the second solution, the third solution, the fourth solution, the xanthine solution and the xanthine solution after being left at room temperature for 1 hour in the preparation process of the embodiment were observed. The results are as follows Figure 1 shown.
[0034] Depend on Figure 1 b and Figure 1 c It can be seen that when ammonia and ethanol are added to xanthine powder, xanthine is dissolved. Figure 1 d It can be seen that the addition of a small amount of hydrochloric acid does not cause the xanthine solution to precipitate. Figure 1It can be seen that the solvent provided by the present invention can effectively dissolve xanthine, making the solution clear and stable without particulate matter, and the needle will not be blocked when drawn with a syringe. Figure 1 f It can be seen that after being placed at room temperature for 1 hour, the xanthine solution is still a clear liquid.
[0035] Example 2: Preparation of xanthine solution
[0036] The neutral solution is composed of 20% ammonia water, 20% ethanol, 20% hydrochloric acid and deionized water in a volume ratio of 12.5:2.5:5:80.
[0037] Place 2 μg of xanthine powder in an EP tube, and sequentially add 12.5 μL of 20% ammonia water, 2.5 μL of 20% ethanol, 5 μL of 20% hydrochloric acid, and 800 μL of deionized water to the xanthine solution, mix well, and obtain a xanthine solution with a concentration of 2 μg / mL. When preparing the xanthine solution, tap the bottom of the tube after each addition of solvent to allow the xanthine powder to fully dissolve.
[0038] Depend on Figure 2 It can be seen that the neutral solvent composed of ammonia water, ethanol, hydrochloric acid and deionized water is used to dissolve xanthine powder, and the obtained xanthine solution is clear and stable without particulate matter.
[0039] Example 3: Preparation of xanthine solution
[0040] The neutral solution is composed of 20% ammonia water, 20% ethanol, 20% hydrochloric acid and deionized water in a volume ratio of 12:2:4:82.
[0041] Place 1 μg of xanthine powder in an EP tube, and sequentially add 12 μL of 20% ammonia water, 2 μL of 20% ethanol, 4 μL of 20% hydrochloric acid, and 820 μL of deionized water to the xanthine solution. Mix well to obtain a xanthine solution with a concentration of 1 μg / mL. When preparing the xanthine solution, tap the bottom of the tube after each addition of solvent to allow the xanthine powder to fully dissolve.
[0042] Example 4: Preparation of xanthine solution
[0043] The neutral solution is composed of 20% ammonia water, 20% ethanol, 20% hydrochloric acid and deionized water in a volume ratio of 13:3:6:78.
[0044] Place 1 μg of xanthine powder in an EP tube, and sequentially add 13 μL of 20% ammonia water, 3 μL of 20% ethanol, 6 μL of 20% hydrochloric acid, and 780 μL of deionized water to the xanthine solution. Mix well to obtain a xanthine solution with a concentration of 1 μg / mL. When preparing the xanthine solution, tap the bottom of the tube after each addition of solvent to allow the xanthine powder to fully dissolve.
[0045] Comparative Example 1: Preparation of xanthine solution
[0046] 2 μg of xanthine powder was placed in an EP tube, and 1000 μL of deionized water was added, and the mixture was mixed by ultrasonic treatment to obtain a xanthine solution. Figure 3 As shown by Figure 3 It can be seen that xanthine precipitates in granular form. Even with ultrasonic treatment, the precipitated xanthine particles cannot be completely dissolved, which makes the stability of the solution poor and easily causes the syringe to be blocked.
[0047] Comparative Example 2:
[0048] 2 μg of xanthine powder was placed in an EP tube, and 1000 μL of 20% ethanol was added and mixed to obtain a xanthine suspension. The dissolution of the xanthine suspension prepared in this comparative example is shown in FIG. Figure 4 As shown by Figure 4 It can be seen that when ethanol is used as a solvent, undissolved xanthine exists in the xanthine suspension formed, and the obtained xanthine suspension is acidic.
[0049] Comparative Example 3:
[0050] 2 μg of xanthine powder was placed in an EP tube, 1000 μL of 20% hydrochloric acid was added, and mixed to obtain a xanthine suspension. The dissolution of the xanthine suspension prepared in this comparative example is shown in FIG. Figure 5 As shown by Figure 5 It can be seen that when hydrochloric acid is used as a solvent, undissolved xanthine exists in the xanthine suspension formed, and the obtained xanthine suspension is acidic.
[0051] Comparative Example 4:
[0052] 2 μg of xanthine powder was placed in an EP tube, and 1000 μL of 20% ammonia water was added and mixed to obtain a xanthine suspension. The dissolution of the xanthine suspension prepared in this comparative example is shown in FIG. Figure 6 As shown by Figure 6 It can be seen that when ammonia water is used as a solvent, undissolved xanthine exists in the xanthine suspension formed, and the obtained xanthine suspension is alkaline.
[0053] Test Example 1: Safety Testing
[0054] The xanthine solution prepared in Example 1 was mixed with deionized water to prepare xanthine solutions with concentrations of 0.1 mg / L, 1.0 mg / L, 10 mg / L, 20 mg / L and 100 mg / L; xanthine solutions of different concentrations were placed in culture dishes as test groups, and a blank control group was set up. The blank control group was a solution mixed with ammonia water, ethanol, hydrochloric acid and deionized water in a volume ratio of 12.5:2.5:5:80, ensuring that the volume of the solution in the culture dishes in the test group and the blank control group was the same; soybean seeds were placed in culture dishes, 20 soybean seeds were placed in each culture dish, and cultured at 25°C and dim light for 7 days. The germination rates of the soybean seeds in the test group and the blank control group were measured. The results are as follows: Figure 7 shown.
[0055] Seeds with the same growth potential in each treatment group were selected and placed in 50g of vermiculite for tissue culture bottle culture experiment. The culture conditions of the tissue culture bottle culture experiment were: room temperature 25℃. After 30 days of culture, soybeans from each treatment group were collected to observe the growth-promoting effect of different concentrations of xanthine solution on soybeans. The results are as follows: Figure 8 shown.
[0056] Depend on Figure 7 It can be seen that under the treatment of 0.1 mg / L xanthine solution, the germination rate of soybean seeds was 65%; under the treatment of 1.0 mg / L xanthine solution, the germination rate of soybean seeds was 55%; under the treatment of 10 mg / L xanthine solution, the germination rate of soybean seeds was 55%; under the treatment of 20 mg / L xanthine solution, the germination rate of soybean seeds was 95%; under the treatment of 100 mg / L xanthine solution, the germination rate of soybean seeds was 5%; in the blank treatment group, only neutral solvent was used to treat soybean seeds, and the germination rate of soybean seeds was 66%. It can be seen that the use of neutral solution treatment alone did not limit seed germination and soybean growth and development, showing significant safety.
[0057] Depend on Figure 8 It can be seen that only using neutral solvent to treat soybeans has a certain promoting effect on the growth, development and morphological structure of soybeans. 0.1-100 mg / L xanthine solution has a certain degree of influence on the growth, development and morphological structure of soybeans, especially 20 mg / L xanthine solution has a significant promoting effect on the growth, development and morphological structure of soybeans.
[0058] Test Example 2: Soil Test
[0059] In this test case, farmland soil and coastal saline-alkali soil were selected as test soils, and soil tests were carried out. The farmland soil was taken from the agronomy experimental field of Shandong Agricultural University, and the coastal saline-alkali soil was taken from Huangdian Village, Kenli District, Dongying City, Shandong Province.
[0060] After mixing deionized water and the xanthine solution prepared in Example 2, a xanthine solution with a concentration of 20 mg / L was obtained. 20 mg / L of xanthine was added to farmland soil and coastal saline-alkali soil as the test group, and the amount of xanthine solution added was 250 mL. A blank control group was also set up, and the blank control group was not added with xanthine solution to farmland soil and coastal saline-alkali soil. The soil pH value of different soils in the test group and the blank treatment group was detected. The soil pH value was detected using a soil pH meter for a solution with a soil-water ratio of 1:2.5. The results are as follows Fig. 9 shown.
[0061] Depend on Fig. 9 As shown, in farmland soil, the soil pH is 7.87 after adding xanthine solution, and the soil pH is 7.89 after not adding xanthine solution; in coastal saline-alkali soil, the soil pH is 8.16 after adding xanthine solution, and the soil pH is 8.13 after not adding xanthine solution. It can be seen that the xanthine soil prepared by the present invention does not change the soil pH, which can greatly ensure the effectiveness and stability of soil experiments.
[0062] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims
1. A neutral solvent for dissolving xanthine, characterized in that, The neutral solvent is composed of ammonia water, ethanol, hydrochloric acid and deionized water in a volume ratio of (12-13): (2-3): (4-6): (78-82).
2. The neutral solvent for dissolving xanthine as claimed in claim 1, characterized in that The mass fraction of the hydrochloric acid is 15-25%, the mass fraction of the ammonia water is 15-25%, and the mass fraction of the ethanol is 15-25%.
3. Use of the neutral solvent described in claim 1 or 2 in the preparation of xanthine solution.
4. A xanthine solution, characterized in that Prepared by the following method: Add ammonia water, ethanol, hydrochloric acid and deionized water to xanthine powder in sequence and mix well to obtain xanthine solution.
5. The xanthine solution as claimed in claim 4, characterized in that The added amount ratio of xanthine powder, ammonia water, ethanol, hydrochloric acid and deionized water is (1-2) g: (12-13) L: (2-3) L: (4-6) L: (78-82) L.
6. Use of the xanthine solution according to claim 4 or 5 in maintaining soil pH stability.