Method for quantitatively detecting 2, 4, 6-trihydroxyacetophenone
By utilizing the 'K2S2O8-Na2S2O3-CuCl2-NH4OH' pH clock system and the linear relationship between induction time and 2,4,6-trihydroxyacetophenone concentration, the problems of complexity and high cost of existing detection methods are solved, and a simple and highly accurate quantitative detection is achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- ANHUI UNIV
- Filing Date
- 2026-01-31
- Publication Date
- 2026-05-08
AI Technical Summary
Existing methods for detecting 2,4,6-trihydroxyacetophenone are complex and costly, lacking efficient and easy-to-operate quantitative analysis techniques.
A pH clock system of 'K2S2O8-Na2S2O3-CuCl2-NH4OH' was used as the detection solution. By recording the pH change over time, a working curve was established for quantitative detection based on the linear relationship between induction time and the concentration of 2,4,6-trihydroxyacetophenone.
It achieves highly accurate quantitative detection in the concentration range of 2.5×10-4 mol/L to 7.5×10-4 mol/L, and is simple to operate and low in cost.
Smart Images
Figure CN121994884A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to an analytical detection method, specifically, the establishment of a "K2S2O8-Na2S2O3-CuCl2-NH4OH" clock system. Based on the different responses of this system to different concentrations of 2,4,6-trihydroxyacetophenone, i.e., the different induction times, a quantitative analysis method for 2,4,6-trihydroxyacetophenone is realized, which belongs to the field of analytical chemistry. Background Technology
[0002] 2,4,6-Trihydroxyacetophenone, with the structure shown in Formula (I), appears as a pale yellow crystalline powder and is soluble in ethanol, ether, chloroform, ethyl acetate, and hot water. 2,4,6-Trihydroxyacetophenone is the glycosidic aglycone of acetophenone glycosides from Curcuma longa, and it has cholesterol-lowering effects, enhancing the activity of cholesterol 7α-hydroxylase. 2,4,6-Trihydroxyacetophenone also stimulates bile secretion through multidrug resistance-associated protein 2. Therefore, it is commonly used as a pharmaceutical intermediate and in the synthesis of novel antioxidants such as flavanones.
[0003] Current methods for detecting 2,4,6-trihydroxyacetophenone include high-performance liquid chromatography (HPLC), ultraviolet-visible spectroscopy (UV-Vis), and proton nuclear magnetic resonance (NMR) spectroscopy. However, these methods are complex and costly. The quantitative analysis method for 2,4,6-trihydroxyacetophenone described in this invention is characterized by high accuracy, ease of operation, and convenience.
[0004]
[0005] The structure of structural formula (Ⅰ) 2,4,6-trihydroxyacetophenone Summary of the Invention
[0006] This invention aims to provide a novel quantitative detection method for 2,4,6-trihydroxyacetophenone, specifically a method for quantitative detection of 2,4,6-trihydroxyacetophenone using a pH clock system of "K2S2O8-Na2S2O3-CuCl2-NH4OH" as the detection solution. This method is based on a standard curve (working curve) method developed using the sensitive response of this pH clock system to 2,4,6-trihydroxyacetophenone. Specifically, the "K2S2O8-Na2S2O3-CuCl2-NH4OH" pH clock reaction system is used as the detection solution, and the pH change over time is recorded. When the pH clock reaction begins, equal volumes of a series of 2,4,6-trihydroxyacetophenone sample solutions of different concentrations are added to the pH clock system. The quantitative detection of the 2,4,6-trihydroxyacetophenone sample is achieved based on the different induction times generated by the system when the concentration of the sample solution in the pH clock system varies. The induction time is the time required for the pH to stabilize after the start of the pH clock system reaction. A working curve was established based on the relationship between the concentration of 2,4,6-trihydroxyacetophenone in a pH clock system and the induction time. The x-axis represents the concentration of 2,4,6-trihydroxyacetophenone in the pH clock system, and the y-axis represents the induction time t. When the concentration of 2,4,6-trihydroxyacetophenone in the system is 2.5 × 10⁻⁶, a working curve is established. -4 mol / L to 7.5×10 -4 When the concentration is between mol / L, the induction time t has a linear relationship with the concentration of 2,4,6-trihydroxyacetophenone, which allows for the quantitative detection of 2,4,6-trihydroxyacetophenone in the sample.
[0007] The difference between this quantitative detection method and the prior art is that the present invention uses the "K2S2O8-Na2S2O3-CuCl2-NH4OH" pH clock system as the detection solution, and the different responses of this system to different concentrations of 2,4,6-trihydroxyacetophenone, i.e., different induction times, to achieve quantitative analysis of 2,4,6-trihydroxyacetophenone.
[0008] The detectable concentration range of 2,4,6-trihydroxyacetophenone in the detection solution (pH clock system) is 2.5 × 10⁻⁶. -4 mol / L to 7.5×10 -4 mol / L.
[0009] When 2,4,6-trihydroxyacetophenone is detected in the detection solution (pH clock system), the temperature of the pH clock system is controlled at any specific temperature within the range of 20-30 °C.
[0010] Using the pH clock system described above, the detectable concentration range of 2,4,6-trihydroxyacetophenone is the optimal concentration range determined experimentally. Within this concentration range, the induction time responds well to changes in the concentration of 2,4,6-trihydroxyacetophenone, exhibiting a large linear correlation coefficient. Furthermore, the concentration ranges of each component in the detection solution (pH clock system) are shown in Table 1, and the optimal concentrations of the detection solution (pH clock system) obtained through multiple experiments are shown in Table 2. Table 1: Concentration range of each component in the pH clock system
[0011] Table 2: Optimal concentrations of each component in the pH clock system
[0012] The specific experimental steps are as follows: 1. Prepare the test solution (pH clock system) according to the concentration range specified in Table 1, and control its temperature at any specific temperature within the range of 20-30 ℃. Insert the prepared working electrode (pH composite electrode, Leici, E-331) into the solution. Connect the other end of the working electrode to the computer via a potential / temperature / pH integrated tester (Jiaxing Disheng Electronic Technology Co., Ltd., ZHFX-595). After opening the chemical signal acquisition and analysis program on the computer and setting the acquisition time and sampling speed, quickly click the start button to monitor the pH of the solution. The computer records the collected pH change curve over time, i.e., the pH clock spectrum. When a substance needs to be detected, add the analyte quickly at the same time as the pH clock system reaction begins, and record the pH change over time in the same manner.
[0013] The basic parameters of a pH clock spectrum include: Induction time: The time required for the pH clock system reaction to reach pH stability.
[0014] Establish a working curve to show the relationship between the concentration of 2,4,6-trihydroxyacetophenone in the detection solution and pH induction time. A series of low-concentration 2,4,6-trihydroxyacetophenone solutions were prepared using ethanol as the solvent as sample solutions. Simultaneously with the start of the pH clock system reaction, 100 μL of each of these sample solutions of different concentrations was added to 40 mL of the pH clock system using a pipette, resulting in a 2.5 × 10⁻⁶ ppm concentration of 2,4,6-trihydroxyacetophenone in the system. -4 mol / L - 7.5 × 10 -4 The pH clock system response changes within the range of mol / L; the induction time is denoted as t. The induction time t varies depending on the concentration of 2,4,6-trihydroxyacetophenone in the system. A graph is plotted with the concentration of 2,4,6-trihydroxyacetophenone in the system on the x-axis and t on the y-axis. When the concentration of 2,4,6-trihydroxyacetophenone in the system is between 2.5 × 10⁻⁶ mol / L... -4 mol / L - 7.5 × 10 -4 When the concentration is between mol / L, the induction time t of the pH clock system has a linear relationship with the concentration of 2,4,6-trihydroxyacetophenone, and the working curve is obtained.
[0015] Quantitative detection of 2,4,6-trihydroxyacetophenone When a test sample of unknown concentration is added to the pH clock system at the start of the pH clock system reaction, the induction time (t) of the corresponding pH clock system can be measured. Based on the relationship between t and concentration on the working curve, the concentration of 2,4,6-trihydroxyacetophenone in the detection system can be obtained, and then the concentration of 2,4,6-trihydroxyacetophenone in the test sample can be calculated. Attached Figure Description
[0016] Figure 1 This is a graph showing the change in pH value of the detection solution (pH clock system) over time when no sample to be tested was added, as described in Example 1.
[0017] Figure 2 In Example 1, 2.5 × 10 -4 After adding mol / L 2,4,6-trihydroxyacetophenone, the pH value of the solution (pH clock system) was measured as a function of time.
[0018] Figure 3 In Example 1, 3.75 × 10⁻⁶ was added. -4 After adding mol / L 2,4,6-trihydroxyacetophenone, the pH value of the solution (pH clock system) was measured as a function of time.
[0019] Figure 4 This is the working curve of pH induction time t versus 2,4,6-trihydroxyacetophenone concentration in Example 1.
[0020] Figure 5 This is a graph showing the change in pH value of the detection solution (pH clock system) over time when no sample to be tested was added, as shown in Example 2.
[0021] Figure 6 In Example 2, 5×10 -4 After adding mol / L 2,4,6-trihydroxyacetophenone, the pH value of the solution (pH clock system) was measured as a function of time.
[0022] Figure 7 In Example 2, 6.25 × 10 -4 After adding mol / L 2,4,6-trihydroxyacetophenone, the pH value of the solution (pH clock system) was measured as a function of time.
[0023] Figure 8 This is the working curve of pH induction time t versus 2,4,6-trihydroxyacetophenone concentration in Example 2.
[0024] Figure 9 This is a graph showing the change in pH value of the test solution (pH clock system) over time when no sample to be tested was added, as shown in Example 3.
[0025] Figure 10 In Example 3, 6.25 × 10 -4 After adding mol / L 2,4,6-trihydroxyacetophenone, the pH value of the solution (pH clock system) was measured as a function of time.
[0026] Figure 11In Example 3, 7.5 × 10⁻⁶ was added. -4 After adding mol / L 2,4,6-trihydroxyacetophenone, the pH value of the solution (pH clock system) was measured as a function of time.
[0027] Figure 12 This is the working curve of pH induction time t versus 2,4,6-trihydroxyacetophenone concentration in Example 3. Detailed Implementation Example 1
[0028] A pH clock system using K₂S₂O₈-Na₂S₂O₃-CuCl₂-NH₄OH as the substrate was applied as the detection solution for the quantitative analysis of 2,4,6-trihydroxyacetophenone. Equal volumes of 2,4,6-trihydroxyacetophenone sample solutions of different concentrations were added to the pH clock system to establish a working curve (e.g., a linear relationship) relating the concentration of 2,4,6-trihydroxyacetophenone in the detection system to the induction time. This allowed for the detection of 2,4,6-trihydroxyacetophenone in the pH clock system, and the subsequent calculation of the concentration of 2,4,6-trihydroxyacetophenone in the test sample.
[0029] (1) Preparation of detection solution First, prepare 0.1 mol / L K2S2O8 solution, 0.0048 mol / L Na2S2O3 solution, 0.01 mol / L CuCl2 solution and 0.01 mol / L NH3·H2O solution (NH4OH solution) using distilled water. Add 10 mL of 0.01 mol / L NH4OH solution, 24.6 mL of 0.0048 mol / L Na2S2O3 solution, 0.5 mL of 0.01 mol / L CuCl2 solution, and 4.9 mL of 0.1 mol / L K2S2O8 solution sequentially to a 50 mL beaker to ensure that the concentrations of each component in the "K2S2O8-Na2S2O3-CuCl2-NH4OH" pH clock system are 0.0025 mol / L NH4OH, 0.002952 mol / L Na2S2O3, 0.000125 mol / L CuCl2, and 0.01225 mol / L K2S2O8, for a total volume of 40 mL. The temperature is controlled at 25 ℃.
[0030] Meanwhile, using ethanol as a solvent, a series of 2,4,6-trihydroxyacetophenone sample solutions of different concentrations were prepared.
[0031] (2) Obtain pH clock spectrum The pH value of the prepared test solution over time was plotted by a computer equipped with a chemical signal acquisition and analysis program (without the test sample added). For example... Figure 1As shown, a pH induction time of 135 s was used as a blank control. Two additional test solutions with the same component concentrations as the above-mentioned test solutions were prepared. For one of these solutions, at the start of the reaction, 100 μL of 0.1 mol / L 2,4,6-trihydroxyacetophenone sample solution was added to a 40 mL pH clock system, resulting in a 2.5 × 10⁻⁶ concentration of 2,4,6-trihydroxyacetophenone in the test solution. -4 The addition of 2,4,6-trihydroxyacetophenone at a concentration of mol / L prolonged the induction time to 164 s. Figure 2 As shown; for the other group, at the start of the reaction, 100 μL of 0.15 mol / L 2,4,6-trihydroxyacetophenone sample solution was added to a 40 mL pH clock system, so that the concentration of 2,4,6-trihydroxyacetophenone in the detection solution was 3.75 × 10⁻⁶. -4 The addition of 2,4,6-trihydroxyacetophenone at a concentration of mol / L changed the induction time to 192 s. Figure 3 As shown. Figure 2 , Figure 3 This confirmed that different concentrations of 2,4,6-trihydroxyacetophenone in the detection solution led to different induction times in the pH clock system. When the concentration of 2,4,6-trihydroxyacetophenone in the detection system was 2.5 × 10⁻⁶, the induction time was significantly different. -4 mol / L - 7.5 × 10 -4 When the concentration is between mol / L, different concentrations can lead to different induction times in the pH clock system, which can be observed.
[0032] (3) Quantitative detection A working curve was established based on the relationship between the concentration of 2,4,6-trihydroxyacetophenone in the detection system and the induction time, such as... Figure 4 As shown, the horizontal axis represents the concentration of 2,4,6-trihydroxyacetophenone in the pH clock system, and the vertical axis represents the induction time t. When the concentration of 2,4,6-trihydroxyacetophenone in the detection system is 2.5 × 10⁻⁶,... -4 mol / L - 7.5 × 10 -4 When the concentration is between mol / L, the induction time has a linear relationship with the concentration of 2,4,6-trihydroxyacetophenone, and the linear equation is t = 212207c + 112.8, R 2 =0.9988. Based on this, quantitative detection of 2,4,6-trihydroxyacetophenone in the sample can be achieved. Example 2
[0033] (1) Preparation of detection solution First, prepare 0.1 mol / L K2S2O8 solution, 0.0048 mol / L Na2S2O3 solution, 0.01 mol / L CuCl2 solution and 0.01 mol / L NH3·H2O solution (NH4OH solution) using distilled water. Add 10.1 mL of 0.01 mol / L NH4OH solution, 24.5 mL of 0.0048 mol / L Na2S2O3 solution, 0.4 mL of 0.01 mol / L CuCl2 solution, and 5 mL of 0.1 mol / L K2S2O8 solution sequentially to a 50 mL beaker to ensure that the concentrations of each component in the "K2S2O8-Na2S2O3-CuCl2-NH4OH" pH clock system are 0.002525 mol / L NH4OH, 0.00294 mol / L Na2S2O3, 0.0001 mol / L CuCl2, and 0.0125 mol / L K2S2O8, for a total volume of 40 mL. The temperature is controlled at 25 °C.
[0034] Meanwhile, using ethanol as a solvent, a series of 2,4,6-trihydroxyacetophenone sample solutions of different concentrations were prepared.
[0035] (2) Obtain pH clock spectrum The pH value of the prepared test solution over time was plotted by a computer equipped with a chemical signal acquisition and analysis program (without the test sample added). For example... Figure 5 As shown, a pH induction time of 137 s was used as a blank control. Two additional test solutions with the same component concentrations as the above test solutions were prepared. For one of these solutions, at the start of the reaction, 100 μL of 0.2 mol / L 2,4,6-trihydroxyacetophenone sample solution was added to a 40 mL pH clock system, resulting in a 2,4,6-trihydroxyacetophenone concentration of 5 × 10⁻⁶ in the test solution. -4 The addition of 2,4,6-trihydroxyacetophenone at a concentration of mol / L prolonged the induction time to 218 s. Figure 6 As shown; for the other group, at the start of the reaction, 100 μL of 0.25 mol / L 2,4,6-trihydroxyacetophenone sample solution was added to a 40 mL pH clock system, so that the concentration of 2,4,6-trihydroxyacetophenone in the detection solution was 6.25 × 10⁻⁶. -4 The addition of 2,4,6-trihydroxyacetophenone at a concentration of mol / L changed the induction time to 243 s. Figure 7 As shown. Figure 6 , Figure 7This confirmed that different concentrations of 2,4,6-trihydroxyacetophenone in the detection solution led to different induction times in the pH clock system. When the concentration of 2,4,6-trihydroxyacetophenone in the detection system was 2.5 × 10⁻⁶, the induction time was significantly different. -4 mol / L - 7.5 × 10 -4 When the concentration is between mol / L, different concentrations can lead to different induction times in the pH clock system, which can be observed.
[0036] (3) Quantitative detection A working curve was established based on the relationship between the concentration of 2,4,6-trihydroxyacetophenone in the detection system and the induction time, such as... Figure 8 As shown, the horizontal axis represents the concentration of 2,4,6-trihydroxyacetophenone in the pH clock system, and the vertical axis represents the induction time t. When the concentration of 2,4,6-trihydroxyacetophenone in the detection system is 2.5 × 10⁻⁶, the value is determined by the induction time t. -4 mol / L - 7.5 × 10 -4 When the concentration is between mol / L, the induction time has a linear relationship with the concentration of 2,4,6-trihydroxyacetophenone, and the linear equation is t = 211200c + 110.2, R 2 = 0.9975. Based on this, quantitative detection of 2,4,6-trihydroxyacetophenone in the sample can be achieved. Example 3
[0037] (1) Preparation of detection solution First, prepare 0.1 mol / L K2S2O8 solution, 0.0048 mol / L Na2S2O3 solution, 0.01 mol / L CuCl2 solution and 0.01 mol / L NH3·H2O solution (NH4OH solution) using distilled water. Add 9.9 mL of 0.01 mol / L NH4OH solution, 24.3 mL of 0.0048 mol / L Na2S2O3 solution, 0.6 mL of 0.01 mol / L CuCl2 solution, and 5.2 mL of 0.1 mol / L K2S2O8 solution sequentially to a 50 mL beaker to ensure that the concentrations of each component in the "K2S2O8-Na2S2O3-CuCl2-NH4OH" pH clock system are 0.002475 mol / L NH4OH, 0.002916 mol / L Na2S2O3, 0.00015 mol / L CuCl2, and 0.013 mol / L K2S2O8, for a total volume of 40 mL. The temperature is controlled at 25 °C.
[0038] Meanwhile, using ethanol as a solvent, a series of 2,4,6-trihydroxyacetophenone sample solutions of different concentrations were prepared.
[0039] (2) Obtain pH clock spectrum The pH value of the prepared test solution over time was plotted by a computer equipped with a chemical signal acquisition and analysis program (without the test sample added). For example... Figure 9 As shown, a pH induction time of 138 s was used as a blank control. Two additional test solutions with the same component concentrations as the above-mentioned test solutions were prepared. For one of these solutions, at the start of the reaction, 100 μL of a 0.25 mol / L 2,4,6-trihydroxyacetophenone sample solution was added to a 40 mL pH clock system, resulting in a 2,4,6-trihydroxyacetophenone concentration of 6.25 × 10⁻⁶ in the test solution. -4 The addition of 2,4,6-trihydroxyacetophenone at a concentration of mol / L extended the induction time to 240 s. Figure 10 As shown; for the other group, at the start of the reaction, 100 μL of 0.3 mol / L 2,4,6-trihydroxyacetophenone sample solution was added to a 40 mL pH clock system, so that the concentration of 2,4,6-trihydroxyacetophenone in the detection solution was 7.5 × 10⁻⁶. -4 The addition of 2,4,6-trihydroxyacetophenone at a concentration of mol / L changed the induction time to 269 s. Figure 11 As shown. Figure 10 , Figure 11 This confirmed that different concentrations of 2,4,6-trihydroxyacetophenone in the detection solution led to different induction times in the pH clock system. When the concentration of 2,4,6-trihydroxyacetophenone in the detection system was 2.5 × 10⁻⁶, the induction time was significantly different. -4 mol / L - 7.5 × 10 -4 When the concentration is between mol / L, different concentrations can lead to different induction times in the pH clock system, which can be observed.
[0040] (3) Quantitative detection A working curve was established based on the relationship between the concentration of 2,4,6-trihydroxyacetophenone in the detection system and the induction time, such as... Figure 12 As shown, the horizontal axis represents the concentration of 2,4,6-trihydroxyacetophenone in the pH clock system, and the vertical axis represents the induction time t. When the concentration of 2,4,6-trihydroxyacetophenone in the detection system is 2.5 × 10⁻⁶, the value is determined by the induction time t. -4 mol / L - 7.5 × 10 -4 When the concentration is between mol / L, the induction time has a linear relationship with the concentration of 2,4,6-trihydroxyacetophenone, and the linear equation is t = 214401c + 109.4, R. 2 = 0.9985. Based on this, quantitative detection of 2,4,6-trihydroxyacetophenone in the sample can be achieved.
Claims
1. A method for the quantitative detection of 2,4,6-trihydroxyacetophenone, characterized in that: A solution of the sample 2,4,6-trihydroxyacetophenone was prepared using ethanol as a solvent. The pH clock reaction system "K2S2O8-Na2S2O3-CuCl2-NH4OH" was used as the detection solution, and the pH of the clock system was recorded over time. The pH clock system temperature is controlled at any specific temperature within the range of 20-30 °C. When the pH clock reaction begins, equal volumes of a series of test sample solutions of different concentrations of 2,4,6-trihydroxyacetophenone are added to the pH clock system. The quantitative detection of the test sample is achieved based on the different induction times generated by the system when the concentration of the test solution in the clock system varies. The induction time is the time required from the start of the pH clock system reaction to pH stabilization. A working curve was established based on the relationship between the concentration of 2,4,6-trihydroxyacetophenone in the test solution and the induction time in the pH clock system. The x-axis represents the concentration of 2,4,6-trihydroxyacetophenone in the test solution in the pH clock system, and the y-axis represents the induction time t. When the concentration of 2,4,6-trihydroxyacetophenone in the system is 2.5 × 10⁻⁶, a working curve is established. -4 mol / L to 7.5×10 -4 When the concentration is between mol / L, the induction time t has a linear relationship with the concentration of 2,4,6-trihydroxyacetophenone, thereby enabling the quantitative detection of 2,4,6-trihydroxyacetophenone in the sample. The molar concentration ranges of each component in the solution were as follows: K2S2O8 0.0115-0.0135 mol / L, Na2S2O3 0.0021-0.0041 mol / L, CuCl2 0.00005-0.00025 mol / L, and NH4OH 0.0015-0.0035 mol / L.
2. The quantitative detection method according to claim 1, characterized in that: The molar concentrations of each component in the solution were: K2S2O8 0.01225 mol / L, Na2S2O3 0.002952 mol / L, CuCl2 0.000125 mol / L, and NH4OH 0.0025 mol / L.
3. The quantitative detection method according to claim 1, characterized in that: The temperature of the pH clock system was controlled at 25 °C when detecting 2,4,6-trihydroxyacetophenone solution.