Method for detecting organophosphate in saline alkali soil and application thereof
By mixing saline-alkali soil samples with diatomaceous earth and internal standard solution, heating and extraction, eluting through columns and using gas chromatography tandem mass spectrometry to detect the organic phosphate content in the extract, the problem of difficulty in detecting multiple organic phosphate esters in saline-alkali soil at the same time is solved, and a simultaneous detection and simplified detection process for 15 types of organophosphate esters is achieved, providing a scientific basis for the evaluation and control of ecosystem risks of saline-alkali soil.
Patent Information
- Application Number
- CN202510007155.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-02
- Publication Date
- 2025-03-21
AI Technical Summary
The prior art is difficult to detect multiple organic phosphate esters in saline-alkali soil at the same time, and the detection methods are complex, making it difficult to apply to risk assessment and control of saline-alkali soil ecosystems.
The saline-alkali soil sample was mixed with diatomaceous earth and internal standard solution, heated and extracted, eluted through column and detected the organophosphate content in the extract using gas chromatography tandem mass spectrometry.
The simultaneous detection of 15 kinds of organic phosphate esters in saline-alkali soil has been achieved, simplified the detection process, and provided a scientific basis for the ecosystem risk assessment of saline-alkali soil and the control of organophosphate esters.
Smart Images
Figure CN119667050A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of soil detection, and in particular to a method for detecting organic phosphate in saline-alkali soil and application thereof. Background Art
[0002] Soil is an important gathering place for organophosphate esters (OPEs). Natural degradation, rainwater erosion and other phenomena can accelerate the migration of OPEs in surface garbage or electronic products into the soil. The type and characteristics of soil are closely related to the concentration and distribution of OPEs. In areas with relatively low population density, relatively few industrial products are used and discarded, and products with OPEs are also relatively less volatilized into the natural environment through wear and tear. At this time, the average concentration of OPEs in the environment is low, usually less than 100ng / g. In waste recycling sites in densely populated cities or population gathering places, the average concentration of OPEs detected in the soil near the garbage accumulation point is often higher. The source analysis of OPEs in soil and actual research results both point to the average concentration of OPEs in soil being positively correlated with the population concentration and economic development level of the region. The concentration of OPEs in sediments in different regions varies greatly, and there are many types. The detection methods of different types of organophosphates are different. The existing technology has not found any method that can simultaneously detect multiple organophosphates in soil.
[0003] Based on this, the present invention is proposed. Summary of the invention
[0004] The purpose of the present invention is to provide a method for detecting organic phosphates in saline-alkali soil and application thereof.
[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:
[0006] The present invention provides a method for detecting organophosphate in saline-alkali soil, comprising the following steps:
[0007] (1) Mixing a saline-alkali soil sample with diatomaceous earth and an internal standard solution, heating and extracting for 4 to 6 minutes, and obtaining an extract;
[0008] (2) passing the extract through a column, eluting, collecting the eluate, blowing it to near dryness with nitrogen, and re-dissolving the organic reagent to obtain a sample to be tested containing an organic phosphate;
[0009] (3) Detecting the content of organophosphates in the sample by gas chromatography-tandem mass spectrometry;
[0010] The chromatographic conditions are as follows: the chromatographic column is SH-Rxi-5SilMS; the mobile phase A is ammonium acetate solution, and the initial concentration of the ammonium acetate solution is 0.08-0.12 wt%; the mobile phase B is methanol; the single injection volume is 5-7 μL; the injection rate is 0.9-1.0 mL / min; the injection temperature is 250-270°C; and the column temperature is 55-65°C.
[0011] The mass spectrometry conditions are as follows: electrospray ion source; ion source temperature of 220-240°C; interface temperature of 290-310°C; acquisition interval of 0.25-0.35s; acquisition mode of multiple reaction selected ion detection.
[0012] Preferably, the mass ratio of the saline-alkali soil sample and diatomaceous earth mixed in step (1) is 8-12 g: 1-3 g;
[0013] The mass volume ratio of the saline-alkali soil sample and the internal standard solution mixed in step (1) is 8-12 g:50 μL.
[0014] Preferably, the internal standard solution in step (1) is a mixture of tributyl phosphate solution, tri(1-chloro-2-propyl) phosphate solution and triphenyl phosphate solution;
[0015] The volume ratio of the tributyl phosphate solution, the tri(1-chloro-2-propyl) phosphate solution and the triphenyl phosphate solution in the internal standard solution is 1-2:1-2:1-2;
[0016] The initial concentrations of the tributyl phosphate solution, tri(1-chloro-2-propyl) phosphate solution and triphenyl phosphate solution in the internal standard solution are independently 90-110 μg / L.
[0017] Preferably, the temperature of the heated extraction in step (1) is 75-85° C.; the pressure of the heated extraction is 9-11 MPa; and the extractant of the heated extraction is a solvent obtained by mixing n-hexane and acetone in a volume ratio of 1-2:1-2.
[0018] Preferably, the column used in step (2) is an amino extraction column; and the elution solvent used in step (2) is a solvent obtained by mixing n-hexane and acetone in a volume ratio of 1-2:1-2.
[0019] Preferably, the organic reagent in step (2) is a solvent obtained by mixing n-hexane and acetone in a volume ratio of 1-2:1-2.
[0020] Preferably, the detected organic phosphate includes one or more of trimethyl phosphate, triethyl phosphate, tripropyl phosphate, triisopropyl phosphate, tributoxyethyl phosphate, tributyl phosphate, triisobutyl phosphate, tri(2-chloroethyl) phosphate, tri(2-chloroisopropyl) phosphate, tri(1,3-dichloroisopropyl) phosphate, triphenyl phosphate, diphenylcresyl phosphate, 3-ethylhexyl diphenyl phosphate, tricresyl phosphate and tri(isopropylbenzene) phosphate.
[0021] The present invention also provides application of the method in preparing a product for detecting organic phosphates in saline-alkali soil.
[0022] The present invention provides a method for detecting organic phosphates in saline-alkali soil and its application. The detection method of the present invention comprises the following steps: (1) mixing a saline-alkali soil sample with diatomaceous earth and an internal standard solution, heating and extracting for 4 to 6 minutes to obtain an extract; (2) passing the extract through a column, eluting, collecting the eluate, blowing it to near dryness with nitrogen, and re-dissolving an organic reagent to obtain a sample to be tested containing organic phosphates; (3) detecting the content of organic phosphates in the sample to be tested by gas chromatography tandem mass spectrometry. The chromatographic conditions are as follows: the chromatographic column is SH-Rxi-5Sil MS; the mobile phase A is ammonium acetate solution, and the initial concentration of the ammonium acetate solution is 0.08-0.12wt%; the mobile phase B is methanol; the single injection volume is 5-7μL; the injection rate is 0.9-1.0mL / min; the injection temperature is 250-270℃; the column temperature is 55-65℃; the mass spectrometry conditions are as follows: electrospray ion source; the temperature of the ion source is 220-240℃; the interface temperature is 290-310℃; the collection interval is 0.25-0.35s; the collection method is multiple reaction selected ion detection. The method according to the present invention can detect the content of 15 kinds of organophosphates in saline-alkali soil, and the detection method is simple, which provides a basis for the detection of organophosphates in saline-alkali soil, and provides a basis for the risk assessment of saline-alkali soil ecosystem and the control of organophosphates in saline-alkali soil. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 Flow chart for organophosphate detection;
[0024] Figure 2 is the standard curve of organophosphate (the left figure is trimethyl phosphate, and the right figure is triethyl phosphate);
[0025] Figure 3 is the standard curve of organophosphate (the left figure is triisobutyl phosphate, the right figure is tripropyl phosphate);
[0026] Figure 4 is the standard curve of organophosphate (the left figure is triisopropyl phosphate, the right figure is tributyl phosphate);
[0027] Figure 5is the standard curve of organophosphate (the left figure is tris(2-chloroethyl) phosphate, and the right figure is tris(2-chloroisopropyl) phosphate);
[0028] Figure 6 is the standard curve of organophosphate (the left figure is tris(1,3-dichloroisopropyl) phosphate, and the right figure is tributoxyethyl phosphate);
[0029] Figure 7 is the standard curve of organophosphate (the left figure is triphenyl phosphate, the right figure is 3-ethylhexyl diphenyl phosphate;
[0030] Figure 8 is the standard curve of organophosphate (the left figure is tricresyl phosphate, the right figure is diphenylcresyl phosphate;
[0031] Fig. 9 is the standard curve of tri(isopropylphenyl) phosphate;
[0032] Fig.10 are the types and concentrations of organophosphates in the soil sample at point A;
[0033] Fig.11 are the types and concentrations of organophosphates in the soil samples at site B;
[0034] Fig.12 are the types and concentrations of organophosphates in the soil sample at point C. DETAILED DESCRIPTION
[0035] The solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be understood as limiting the protection scope of the present invention.
[0036] In the embodiments of the present invention, the organic phosphates are shown in Table 1:
[0037] Table 115 organophosphates and corresponding internal standards
[0038]
[0039] The internal standards in the examples of the present invention are tributyl phosphate (TnBP-d27), tri(1-chloro-2-propyl) phosphate (TCPP-d18) and triphenyl phosphate (TPhP-15).
[0040] Example 1
[0041] Soil samples (15 in total, 5 in each area) were collected around the Liupai Gan (A), Qipai Gan (B) and Tongxin Village (C) of Longxingchang Town in the Yellow River Basin of Fuxing Town, Bayannur City, Inner Mongolia. The soil in this area belongs to saline-alkali land. The specific coordinates of the sampling points are shown in Table 2. Soil samples were collected using a shovel. The weeds on the surface of the sample point were removed with a shovel. Multiple sample points were collected from each point using the plum blossom distribution method. The samples in the same area were mixed to remove larger stones and animal and plant residues. The sample point collection method was as follows: 0 to 100 cm of the film-covered farmland, every 20 cm as a soil layer, 2 bags (bag size: 200 mm × 140 mm), 1.4 kg per bag, were collected from each soil layer as soon as possible and transported back to the laboratory and stored at -20 °C.
[0042] Table 2 Coordinate positions of sampling points
[0043] Point coordinate A 107.962°E, 41.105°N B 108.167°E, 41.052°N C 108.344°E, 41.115°N
[0044] The collected soil was freeze-dried, ground in a mortar, and passed through a 100-mesh sieve to obtain a saline-alkali soil sample.
[0045] 10 g of saline-alkali soil sample was mixed with 2 g of diatomaceous earth and 50 μL of internal standard solution, and loaded into the extraction cell of the accelerated solvent extractor. 40 mL of n-hexane / acetone (volume ratio of 1:1) was used as the extractant. The extraction system pressure was set to 10 MPa, the temperature was 80 ° C, the heating time was 5 min, the static time was 5 min, the flushing volume was 60%, and the cycle was repeated twice to obtain the extract. An amino extraction column (500 mg / 6 mL) was used for solid phase extraction, and the extraction was carried out in sequence. 5mL n-hexane / acetone (volume ratio of 1:1), 5mL n-hexane and 10mL ultrapure water were used to activate the extraction column, and the sample was loaded, and the sample was transferred to the small column with n-hexane (a total of 2mL) in small amounts and multiple times; the small column was eluted with 10mL n-hexane / acetone (volume ratio of 1:1) and the eluent was collected, blown to near dryness with nitrogen, and the volume was fixed to 1mL with n-hexane / acetone (volume ratio of 1:1) to obtain the sample to be tested; the content of organophosphate in the sample to be tested was detected by gas chromatography tandem mass spectrometry. The internal standard solutions included nBP-27 solution, TCPP-18 solution and TPhP-15 solution, the initial concentration of each solution was 100μg / L, and the volume ratio of each solution was 1:1:1.
[0046] Chromatographic parameters: chromatographic column: SH-Rxi-5Sil MS (30m×0.25mm×0.25μm); mobile phase A: ammonium acetate solution, initial concentration of ammonium acetate solution: 0.08-0.12wt%; mobile phase B: methanol; single injection volume: 6μL; injection rate: 0.97mL / min; injection temperature: 260℃; column temperature: 60℃;
[0047] The mass spectrometry conditions are: electrospray ion source; ion source temperature is 230°C; interface temperature is 300°C; acquisition interval is 0.3s; acquisition mode is multiple reaction selected ion detection. The specific detection flow chart is as follows Figure 1 shown.
[0048] The gradient elution program is shown in Table 3.
[0049] Table 3 Gas chromatograph temperature heating program
[0050] rate Temperature(℃) Holding time (min) 0 60.0 2.00 15.00 210.0 2.00 30.00 300.0 10.00
[0051] Example 2
[0052] Drawing of standard curve
[0053] Use methanol solution to dilute the 15 kinds of organophosphate esters (OPEs) described in Table 1 to prepare different gradient standard solutions. Accurately weigh 1.0 mg of 15 kinds of OPEs standards, dissolve them in 1.0 mL of methanol solution to obtain 15 kinds of OPEs initial standard solutions, and then divide them into five parts, and dilute them with methanol solution to obtain 20, 50, 100, 200, 300 μg / L standard solutions, which are stored at -20°C.
[0054] The standard solution was processed according to the method of Example 1, 0.5 mL of the standard solution was equivalent to 0.5 g of the saline-alkali soil sample of Example 1, and the detection method was carried out according to the parameters of the gas chromatography tandem mass spectrometry method of Example 1. The peak area was measured, and the standard curves of 15 kinds of organophosphates were obtained by the least squares method ( Figures 2 to 9 ), the horizontal axis is the concentration, and the vertical axis is the peak area. The standard curve is shown in Table 4.
[0055] Table 4 Standard curve
[0056]
[0057]
[0058] Example 3
[0059] The type of OPEs represented by the characteristic peak is determined according to the specific peak time, and the concentration of each organophosphate is obtained by comparing the obtained peak area with the standard curve. The types of OPEs and the concentrations of organophosphates detected at the three distribution points in Example 1 are shown in FIG. Figures 10-12 shown.
[0060] Figures 10-12As shown, a total of 8 organophosphates were detected at the three sites, while TMP, TEP, TPrP, TiBP, CDPP, EHDPP and TCrP were not detected. Among the detected OPEs, TBEP had the highest average concentration, with concentrations of 15.81 ng / g, 10.07 ng / g and 49.79 ng / g at sites A, B and C, respectively, followed by TCHP, with average concentrations of 9.66 ng / g, 2.30 ng / g and 2.01 ng / g, dry weight, respectively.
[0061] As can be seen from the above embodiments, the present invention provides a method for detecting organic phosphates in saline-alkali soil and its application. The detection method of the present invention comprises the following steps: (1) mixing a saline-alkali soil sample with diatomaceous earth and an internal standard solution, heating and extracting for 4 to 6 minutes to obtain an extract; (2) passing the extract through a column, eluting, collecting the eluate, blowing it to near dryness with nitrogen, and re-dissolving the organic reagent to obtain a sample to be tested containing organic phosphates; (3) detecting the content of organic phosphates in the sample to be tested by gas chromatography-tandem mass spectrometry. The chromatographic conditions are as follows: the chromatographic column is SH-Rxi-5Sil MS; the mobile phase A is ammonium acetate solution, and the initial concentration of the ammonium acetate solution is 0.08-0.12wt%; the mobile phase B is methanol; the single injection volume is 5-7μL; the injection rate is 0.9-1.0mL / min; the injection temperature is 250-270℃; the column temperature is 55-65℃; the mass spectrometry conditions are as follows: electrospray ion source; the temperature of the ion source is 220-240℃; the interface temperature is 290-310℃; the collection interval is 0.25-0.35s; the collection method is multiple reaction selected ion detection. The method according to the present invention can detect the content of 15 kinds of organophosphates in saline-alkali soil, and the detection method is simple, which provides a basis for the detection of organophosphates in saline-alkali soil, and provides a basis for the risk assessment of saline-alkali soil ecosystem and the control of organophosphates in saline-alkali soil.
[0062] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A method for detecting organophosphates in saline-alkali soil, characterized in that: The steps include: (1) Mixing a saline-alkali soil sample with diatomaceous earth and an internal standard solution, heating and extracting for 4 to 6 minutes, and obtaining an extract; (2) passing the extract through a column, eluting, collecting the eluate, blowing it to near dryness with nitrogen, and re-dissolving the organic reagent to obtain a sample to be tested containing an organic phosphate; (3) Detecting the content of organophosphates in the sample by gas chromatography-tandem mass spectrometry; The chromatographic conditions are as follows: the chromatographic column is SH-Rxi-5SilMS; the mobile phase A is ammonium acetate solution, and the initial concentration of the ammonium acetate solution is 0.08-0.12 wt%; the mobile phase B is methanol; the single injection volume is 5-7 μL; the injection rate is 0.9-1.0 mL / min; the injection temperature is 250-270°C; and the column temperature is 55-65°C. The mass spectrometry conditions are as follows: electrospray ion source; ion source temperature of 220-240°C; interface temperature of 290-310°C; acquisition interval of 0.25-0.35s; acquisition mode of multiple reaction selected ion detection.
2. The method according to claim 1, characterized in that The mass ratio of the saline-alkali soil sample and diatomaceous earth mixed in step (1) is 8-12 g:1-3 g; The mass volume ratio of the saline-alkali soil sample and the internal standard solution mixed in step (1) is 8-12 g:50 μL.
3. The method according to claim 2, characterized in that The internal standard solution in step (1) is a mixture of tributyl phosphate solution, tri(1-chloro-2-propyl) phosphate solution and triphenyl phosphate solution; The volume ratio of the tributyl phosphate solution, the tri(1-chloro-2-propyl) phosphate solution and the triphenyl phosphate solution in the internal standard solution is 1-2:1-2:1-2; The initial concentrations of the tributyl phosphate solution, tri(1-chloro-2-propyl) phosphate solution and triphenyl phosphate solution in the internal standard solution are independently 90-110 μg / L.
4. The method according to claim 3, characterized in that The temperature of the heating extraction in step (1) is 75-85° C.; the pressure of the heating extraction is 9-11 MPa; and the extractant of the heating extraction is a solvent obtained by mixing n-hexane and acetone in a volume ratio of 1-2:1-2.
5. The method according to claim 4, characterized in that The column used in step (2) is an amino extraction column; the elution solvent used in step (2) is a solvent obtained by mixing n-hexane and acetone in a volume ratio of 1-2:1-2.
6. The method according to claim 5, characterized in that The organic reagent in step (2) is a solvent obtained by mixing n-hexane and acetone in a volume ratio of 1-2:1-2.
7. The method according to any one of claims 1 to 6, characterized in that: The detected organic phosphates include one or more of trimethyl phosphate, triethyl phosphate, tripropyl phosphate, triisopropyl phosphate, tributoxyethyl phosphate, tributyl phosphate, triisobutyl phosphate, tri(2-chloroethyl) phosphate, tri(2-chloroisopropyl) phosphate, tri(1,3-dichloroisopropyl) phosphate, triphenyl phosphate, diphenylcresyl phosphate, 3-ethylhexyldiphenyl phosphate, tricresyl phosphate and tri(isopropylbenzene) phosphate.
8. Use of the method according to any one of claims 1 to 7 in the preparation of a product for detecting organophosphates in saline-alkali soils.