CN113916815A - Method for measuring content of ammonia nitrogen in seawater by using Nashin's reagent spectrophotometry - Google Patents
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Abstract
The invention provides a method for measuring the content of ammonia nitrogen in seawater by a nano-grade reagent spectrophotometry. The method finds the optimal addition amount of the potassium sodium tartrate within a wider salinity range by adjusting the concentration and the volume of the potassium sodium tartrate according to the salinity of a sample to be detected, and can eliminate Ca in the seawater sample by only using a potassium sodium tartrate solution as a masking agent2+、Mg2+、Fe2+And Mn2+And the interference of impurity ions enables the NahLo reagent in the seawater to be more stable in color development, and the method is wider in application range. The method does not need reagents such as sodium hydroxide and the like as masking assistance, the consumption of the potassium sodium tartrate and the Nashi reagent is less, the environmental pollution is reduced, and the cost is saved; has the advantages of less interference, simple operation, high speed, high recovery rate, good precision and the like.
Description
Technical Field
The invention relates to the technical field of environmental detection and analysis, in particular to a method for measuring the content of ammonia nitrogen in seawater by a nano-reagent spectrophotometry.
Technical Field
The method for determining ammonia nitrogen in seawater is a common sodium hypobromite oxidation method, and the method principle is that ammonia nitrogen is oxidized into nitrite by hypobromite in an alkaline medium, then the total amount of nitrite nitrogen is measured by a diazo-azo spectrophotometry, and the content of nitrite in a sample is deducted to obtain the concentration of ammonia nitrogen. The method has high requirements on reaction conditions, long testing time, great influence of reagent quality on testing, difficult mastering, large testing error and poor reproducibility, and is not suitable for measuring the ammonia nitrogen content in the mariculture sample with heavy pollution and high organic matter content. The indophenol blue spectrophotometry is also a classic method for measuring ammonia nitrogen in seawater, and the principle of the method is that ammonia oxidation is carried out under the action of hypochlorous acid to generate chloramine, the chloramine reacts with phenol to generate an indophenol blue complex, and a spectrophotometer is used for detecting the absorbance of the complex at 640 nm. However, the method has low sensitivity, long time consumption, complex required reagent and small linear range. In addition, the distillation-titration method, the gas phase molecular absorption spectrometry method and the like can also be used for measuring the ammonia nitrogen in the seawater. However, the above methods are either complicated to operate, require high conditions, are time-consuming, have poor reproducibility, or use expensive instruments, thereby limiting their practical applications.
The principle of the standard analysis method adopted by many countries is that an alkaline solution of mercuric iodide and potassium iodide reacts with ammonia to generate a light red colloidal compound, the compound is strongly absorbed within a range of 410-425 nm, and the light absorption value is in direct proportion to the ammonia nitrogen concentration within a certain range, so that the method is usually used for measuring fresh water and sewage samples. But when the sample to be detected is seawater with higher salt content or contains seawater, Ca in the sample to be detected2+、Mg2+、Fe2+、Mn2+The plasma will react withThe Nash reagent reacts, producing turbidity or precipitation that interferes with the assay. Although a plurality of documents in China improve the method so as to apply the Nashin reagent method to determine the ammonia nitrogen in the seawater sample, the improvement method is found to have poor applicability in the actual operation process, most methods are assisted by adding a sodium hydroxide solution, the dosage is not easy to control, the operation difficulty is great, and the effect is not stable.
Disclosure of Invention
The invention aims to provide a method for measuring the content of ammonia nitrogen in seawater by a nano-grade reagent spectrophotometry, aiming at the defects in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a method for measuring the content of ammonia nitrogen in seawater by a nano-grade reagent spectrophotometry, which is suitable for a seawater sample with salinity not higher than 35ppt, and comprises the following steps:
s1 preparation of solution
Blank solution: ammonia-free seawater;
standard curve solution: preparing standard ammonium use solutions with different concentrations by using the blank solution as a dilution solution to obtain standard curve solutions with different ammonia nitrogen concentrations;
sample solution: taking a sample to be detected, and filtering the sample through a filter membrane to obtain filtrate as the sample solution; if the ammonia nitrogen content of the sample solution is too high, a blank solution can be used as a dilution solution for dilution;
s2, drawing a standard curve
Drawing a standard curve according to the following steps:
(a) respectively taking a certain volume of the standard curve solutions, and placing the standard curve solutions in different test tubes;
(b) adding potassium sodium tartrate solution with certain concentration in a volume ratio of 0.8:1 respectively by taking the volume of the solution of the standard curve as a reference, and uniformly mixing;
(c) adding Nashin's reagent with volume ratio of 0.1:1 respectively with the volume of the standard curve solution as reference, mixing, standing for 15min to stabilize color development;
(d) at the wavelength of 420nm, using a 1cm cuvette, taking a blank standard as a reference solution for zero adjustment, and determining a light absorption value;
(e) drawing a standard curve by taking the light absorption value as a vertical coordinate and the ammonia nitrogen concentration (mg/L) as a horizontal coordinate, and fitting to obtain a standard curve equation;
s3, sample measurement
Transferring sample solution with the same volume after being filtered by a filter membrane according to the standard curve solution, and placing the sample solution in a colorimetric tube; measuring the absorbance of the sample solution according to the steps (b) to (d) in step S2;
s4, calculating the content
And calculating the measured absorbance value of the sample solution by using a standard curve equation to obtain the ammonia nitrogen concentration in the sample solution.
Further, the mass concentration of the potassium sodium tartrate solution is 2%.
Further, in step S1, at least 5 standard curve solutions with different ammonia nitrogen concentrations are prepared from the blank solution.
Further, the ammonia nitrogen concentration of the standard curve solution is 0 ppm-8.0 ppm.
Further, the method has a minimum detection limit of 0.05 mg/L.
Further, the method is suitable for seawater samples with the salinity of 9-35 ppt.
The technical scheme provided by the invention has the beneficial effects that:
(1) the method provided by the invention is that according to the salinity of the sample to be detected, the optimal addition amount of the potassium sodium tartrate in a wider salinity range is found by adjusting the concentration and the volume of the potassium sodium tartrate, and Ca in the seawater sample can be eliminated by only using the potassium sodium tartrate solution as a masking agent2+、Mg2+、Fe2+And Mn2+And due to the interference of impurity ions, the color development of the nano-reagent is more stable, and the method has a wider application range.
(2) Compared with the method for measuring the content of ammonia nitrogen in seawater in the prior literature, the method does not need reagents such as sodium hydroxide and the like as masking assistance, reduces the dosage of potassium sodium tartrate and Navier reagent, reduces the environmental pollution and saves the cost; the use of ammonia-free seawater is matched, so that the influence of background factors such as salt on the absorbance determination is eliminated, the result is stable and reliable, the sensitivity is high, and the reproducibility is good; the operation is quick and simple, and the requirement on test conditions is low; the salinity range of the applicable seawater is 9-35 ppt. Through a standard adding recovery test, the standard adding recovery rate of the method is 97-105%, and the measurement requirement of high-salinity seawater on ammonia nitrogen is completely met.
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FIG. 1 is a graph showing the change of color development stability in ammonia nitrogen determination in a seawater sample;
FIG. 2 is a standard curve diagram for measuring ammonia nitrogen in seawater samples.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be further described with reference to the accompanying drawings and examples.
The invention provides a method for measuring the content of ammonia nitrogen in seawater by a nano-grade reagent spectrophotometry, which aims to eliminate Ca in a seawater sample2+、Mg2+、Fe2+And Mn2+The invention researches and determines the usage amount of potassium sodium tartrate, the usage amount of a nano reagent, the color development stability and the applicable seawater salinity as follows:
(1) determination of the amount of potassium sodium tartrate
0,0.40mL of standard ammonium working solution (50mg/L NH) was accurately aspirated4 +N) in test tubes, diluted to 5.0mL with ammonia-free seawater of various salinity, shaken to form test samples of 0,4.0 ppm. Respectively adding 4mL of potassium sodium tartrate solutions with different concentrations, uniformly mixing, adding 0.5mL of the Nashi reagent, and uniformly mixing. After 15min, whether turbidity or precipitation exists or not is observed, if the turbidity exists, the absorbance value is measured by taking 0ppm as a reference at the wavelength of 420nm when the clarification is finished, and the result is shown in table 1, which shows that 4mL of 2% potassium sodium tartrate is optimal in use amount and is suitable for measuring ammonia nitrogen in seawater samples with the salinity of 9-35 ppt.
Table 1: the absorbance value of the ammonia nitrogen 4ppm sample under different salinity seawater and different concentrations of potassium sodium tartrate (-representing turbidity)
(2) Determination of the amount of Nashi reagent
Accurately aspirate 0,0.40ml of standard ammonium working solution (50mg/L NH)4 +N) in test tubes, diluted to 5.0mL with ammonia-free seawater of various salinity, shaken to form test samples of 0,4.0 ppm. Adding 4mL of 2% potassium sodium tartrate solution, mixing uniformly, adding the sodium natrii reagents with different volumes respectively, and mixing uniformly. After 15min, measuring the absorbance value by taking 0ppm as a reference at a wavelength of 420nm, wherein the result is shown in Table 2, which shows that 0.4-0.6mL of the Nashin reagent is suitable for measuring ammonia nitrogen in the seawater sample with the salinity of 9-35.0ppt, and the method adopts the dosage of 0.5mL of the Nashin reagent in order to facilitate the implementation and operation of workers.
Table 2: nagowski reagent dosage test (representing turbidity) for measuring ammonia nitrogen in seawater with different salinity
(3) Determination of the color stability of the Nassler reagent
A test sample with a certain concentration is prepared, according to the method of the invention, 0ppm is taken as a reference at the wavelength of 420nm, the absorbance value is measured, the reading is started from 0min, the reading is carried out every 5min for the first 1h, and the reading is carried out every 10min after 1h, and the result is shown in figure 1, and the absorbance of the sample is relatively stable (the change is less than 5%) within 10min to 90 min.
In order to verify that the method has high accuracy, good precision, wide linear range and wide applicable salinity range, a plurality of samples with different ammonia nitrogen contents are adopted to verify the method:
the method comprises the following specific analysis steps:
s1 preparation of solution
Blank solution: ammonia-free seawater;
the preparation method of the ammonia-free seawater comprises the following steps: collecting clean seawater, filtering with 0.45 μm filter membrane, adding anhydrous sodium carbonate to alkalize to pH 10, adding ammonia-free water of the same volume, boiling to original volume, adding 1+9 hydrochloric acid to adjust pH 7, adding 1ml chloroform into each liter of ammonia-free seawater, mixing, sealing, and storing in light-resistant reagent bottle. Conductivity and salinity were measured and recorded.
Standard curve solution: accurately aspirate 0, 0.10, 0.20, 0.40, 0.60, 0.80mL of ammonium standard working solution (50mg/L NH)4 +-N) diluting to 5.0mL with ammonia-free seawater in a test tube to prepare a standard curve solution with ammonia nitrogen concentration of 0,1.0,2.0,4.0,6.0,8.0 ppm;
the preparation method of the ammonium standard stock solution comprises the following steps: 3.8190g of high-grade pure ammonium chloride dried at 105 ℃ for more than 2h are accurately weighed and dissolved in ammonia-free water, and the solution is transferred into a 1000ml volumetric flask and diluted to a scale mark. The ammonia nitrogen content of the solution is 1000 mg/L.
Preparation method of ammonium standard use solution: 5.0mL of ammonium standard stock solution was accurately transferred to a 100mL volumetric flask and diluted to the mark with ammonia-free seawater. This solution contained 0.05mg ammonia nitrogen per ml, i.e. 50 ppm.
The preparation method of the Nassner reagent comprises the following steps: weighing 16g of sodium hydroxide, dissolving in 50ml of pure water, and fully cooling to room temperature; further, 7g of potassium iodide and 10g of mercuric iodide were weighed and dissolved in pure water, and the solution was gradually poured into a sodium hydroxide solution under stirring and diluted with pure water to 100 ml.
The preparation method of the potassium sodium tartrate solution (2 percent) comprises the following steps: 2g of potassium sodium tartrate tetrahydrate are weighed and dissolved in 100mL of anhydrous water. If the purity of the potassium sodium tartrate is not enough, impurities such as ammonia nitrogen and the like are contained, turbidity or precipitation can be generated during measurement, analysis is influenced, 5mL of the Nashi reagent can be added after constant volume, the mixture is shaken and then kept stand, the bottom precipitation is removed, and the supernatant is stored as a use solution.
Sample solution: taking a sample to be detected, and filtering the sample through a filter membrane to obtain filtrate as the sample solution;
s2, drawing a standard curve
Drawing a standard curve according to the following steps:
(a) taking 5mL of the standard curve solutions with different ammonia nitrogen concentrations, and respectively placing the standard curve solutions in different colorimetric tubes;
(b) respectively adding 4mL of 2% potassium sodium tartrate solution, and uniformly mixing;
(c) respectively adding 0.5mL of Nashi reagent, mixing, and standing for more than 15 min;
(d) at the wavelength of 420nm, a 1cm cuvette is used, a blank solution of 0ppm is used as a reference solution for zero adjustment, and the light absorption value is measured;
(e) drawing a standard curve by taking the light absorption value as a vertical coordinate and the ammonia nitrogen concentration (mg/L) as a horizontal coordinate, and fitting to obtain a standard curve equation;
s3, sample measurement
5mL of filtered sample solution is transferred and placed in a colorimetric tube; measuring the absorbance of the sample solution according to the steps (b) to (d) in step S2;
s4, calculating the content
And calculating the measured absorbance value of the sample solution by using a standard curve equation to obtain the ammonia nitrogen concentration in the sample solution.
As shown in fig. 2, an ammonia nitrogen standard curve of the seawater is drawn, and the ammonia nitrogen standard curve equation of the seawater is that y is 0.1032x +0.0015, R20.9998. The ammonia nitrogen detection limit of the method is 0.05mg/L, and the linear range is 0.1-8.0 mg/L.
Seawater samples from 5 different sources were subjected to a spiking recovery test: adding a known amount of ammonium standard (NH) to the sample to be tested4 +and-N) using liquid, respectively measuring the ammonia nitrogen content of the sample to be measured and the standard adding sample, independently performing three times of measurement, and calculating the standard adding recovery rate.
The standard recovery rate (%) is (standard sample ammonia nitrogen content-background sample ammonia nitrogen content)/standard recovery rate x 100%
The measurements were performed on 5 seawater samples from different sources, and the relative error was calculated in triplicate. The calculated standard recovery rates are shown in table 3, the standard recovery rate of each group of water samples is 96-105%, the standard deviation is less than 3.05%, the method has good recovery rate and precision, is suitable for measuring ammonia nitrogen in seawater samples, and meets the second part of the ocean monitoring standard GB 17378.2: the data processing and analysis quality control stipulates that the method has high accuracy and good precision.
Table 3: measurement result of standard recovery rate of seawater samples from different sources
The effect of the method for measuring the content of ammonia nitrogen in seawater by using a nano-grade reagent spectrophotometry provided by the invention will be described in detail with reference to the following examples.
Example 1:
and (4) measuring the ammonia nitrogen content in the artificial seawater samples with different salinity.
1. Solution preparation
Preparing artificial seawater: 35.0g of sea salt was weighed out and dissolved in 1000mL of ammonia-free water.
Preparing artificial seawater with different salinity: diluting with ammonia-free water in proportion.
Sample 1: 35.0ppt of ammonia-free seawater and 0.5mg/L of ammonia-containing nitrogen.
Sample 2: 35.0ppt of ammonia-free seawater and 3.0mg/L of ammonia-containing nitrogen.
Sample 3: 35.0ppt of ammonia-free seawater and 6.0mg/L of ammonia-containing nitrogen.
Sample 4: 32.74ppt of ammonia-free seawater and 0.5mg/L of ammonia-containing nitrogen.
Sample 5: 32.74ppt of ammonia-free seawater and 3.0mg/L of ammonia-containing nitrogen.
Sample 6: 32.74ppt of ammonia-free seawater and 6.0mg/L of ammonia-containing nitrogen.
Sample 7: 17.83ppt of ammonia-free seawater and 5.0mg/L of ammonia-containing nitrogen.
Sample 8: 8.91ppt of ammonia-free seawater and 2.0mg/L of ammonia-containing nitrogen.
2. Experimental procedure
(1) Drawing a standard curve: accurately aspirate 0, 0.10, 0.20, 0.40, 0.60 and 0.80mL of ammonium standard use solution (50ppm NH)4 +-N) diluting in test tube with ammonia-free seawater of different salinity until 5.0mL becomes 0,1.0,2.0,4.0,6.0,8.0mg/L standard series, adding 4.0mL of 2% potassium sodium tartrate, and mixing completely. Add Nashi reagent 0.4mL, mix well. Standing for 15min, measuring with spectrophotometer at wavelength of 420nm with 0ppm as referenceAn absorbance value. And drawing a standard curve by taking the absorbance value as a vertical coordinate and the ammonia nitrogen concentration as a horizontal coordinate.
(2) And (3) sample determination: samples with different ammonia nitrogen concentrations are prepared by using ammonia-free seawater and artificial seawater with different salinity respectively. And respectively transferring 5.0mL of water sample into the test tubes, measuring the absorbance according to a standard curve method, and solving the concentration of ammonia nitrogen in the water sample in a standard curve equation.
(3) The test was repeated: the same sample was subjected to three independent determinations under repetitive conditions, and the average value was taken.
The results are shown in Table 4, the relative error between the standard value and the measured value of each concentration is less than + -10%, and the standard deviation is less than 5%, which shows that the accuracy and precision of the method are good; the results of measuring the ammonia nitrogen content in seawater samples with the same ammonia nitrogen concentration and different salinity are consistent, which shows that the method is applicable to seawater samples with different salinity.
Table 4: determination result of ammonia nitrogen content in seawater samples with different salinity
Comparative example 1:
and (3) determining the ammonia nitrogen content in the sample by adopting a national standard sodium hypobromite oxidation method.
As shown in Table 5, it was found that the sodium hypobromite was slightly lower in the measurement value by the oxidation method than the method of the present invention, and the normalized recovery rate was compared with R of the standard curve2The method is slightly worse than the method of the invention, probably because the sodium hypobromite oxidation method has high determination sensitivity, and the high-concentration ammonia nitrogen sample needs to be diluted for multiple times and is easy to introduce errors. Compared with the prior art, the method disclosed by the invention is more convenient and stable to determine the high-concentration ammonia nitrogen sample.
Table 5: the method of the invention is compared with the national standard sodium hypobromite oxidation method
| Linear equation of equations | R2 | Measured value, mg/L | Recovery rate of the added standard | |
| Oxidation of sodium hypobromite | Y＝2.75x+0.0040 | 0.9964 | 1.38 | 115 |
| The method of the invention | Y＝0.1125x-0.0021 | 0.9998 | 2.07 | 105 |
The features of the embodiments and embodiments described herein above may be combined with each other without conflict.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (6)
1. A method for measuring the content of ammonia nitrogen in seawater by a nano reagent spectrophotometry is characterized in that: the method is suitable for seawater samples with salinity not higher than 35ppt, and comprises the following steps:
s1 preparation of solution
Blank solution: ammonia-free seawater;
standard curve solution: preparing standard ammonium use solutions with different concentrations by using the blank solution as a dilution solution to obtain standard curve solutions with different ammonia nitrogen concentrations;
sample solution: taking a sample to be detected, and filtering the sample through a filter membrane to obtain filtrate as the sample solution; if the ammonia nitrogen content of the sample solution is too high, a blank solution can be used as a dilution solution for dilution;
s2, drawing a standard curve
Drawing a standard curve according to the following steps:
(a) respectively taking a certain volume of the standard curve solutions, and placing the standard curve solutions in different test tubes;
(b) adding potassium sodium tartrate solution with certain concentration in a volume ratio of 0.8:1 respectively by taking the volume of the solution of the standard curve as a reference, and uniformly mixing;
(c) adding Nashin's reagent with volume ratio of 0.1:1 respectively with the volume of the standard curve solution as reference, mixing, standing for 15min to stabilize color development;
(d) at the wavelength of 420nm, using a 1cm cuvette, taking a blank standard as a reference solution for zero adjustment, and determining a light absorption value;
(e) drawing a standard curve by taking the light absorption value as a vertical coordinate and the ammonia nitrogen concentration (mg/L) as a horizontal coordinate, and fitting to obtain a standard curve equation;
s3, sample measurement
Transferring sample solution with the same volume after being filtered by a filter membrane according to the standard curve solution, and placing the sample solution in a colorimetric tube; measuring the absorbance of the sample solution according to the steps (b) to (d) in step S2;
s4, calculating the content
And calculating the measured absorbance value of the sample solution by using a standard curve equation to obtain the ammonia nitrogen concentration in the sample solution.
2. The method of claim 1 for measuring the content of ammonia nitrogen in seawater by using a nano-reagent spectrophotometry, which comprises the following steps: the mass concentration of the potassium sodium tartrate solution with a certain concentration is 2 percent, or the change is carried out according to the concentration and the volume of the sample.
3. The method of claim 1 for measuring the content of ammonia nitrogen in seawater by using a nano-reagent spectrophotometry, which comprises the following steps: in step S1, at least 5 standard curve solutions with different ammonia nitrogen concentrations are prepared by the blank solution.
4. The method of claim 3 for the spectrophotometric determination of ammonia nitrogen content in seawater, wherein: the ammonia nitrogen concentration range of the standard curve solution is 0 ppm-8.0 ppm.
5. The method for the spectrophotometric determination of ammonia nitrogen content in seawater according to any of claims 1-4, wherein: the minimum detection limit of the method is 0.05 mg/L.
6. The method for the spectrophotometric determination of ammonia nitrogen content in seawater according to any of claims 1-4, wherein: the method is suitable for seawater samples with the salinity of 9-35 ppt.
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