CN109856123B - Rapid detection reagent and detection method for nitrate - Google Patents
Rapid detection reagent and detection method for nitrate Download PDFInfo
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Abstract
The invention discloses a rapid detection reagent for nitrate and a detection method, wherein the rapid detection reagent for nitrate comprises a reagent I and a reagent II, wherein: the reagent I comprises the following components in percentage by mass: organic acid: 58% -88.5%, cadmium powder: 1% -10%, sulfanilamide: 0.5% -2%, polyvinylpyrrolidone: 10% -30%; the reagent II comprises the following components in percentage by mass: naphthyl ethylenediamine hydrochloride: 2% -6%, organic acid: 94 to 98 percent. The on-site rapid detection method for nitrate comprises the following steps: 1) adding a water sample to be detected into a colorimetric bottle, and then adding the nitrate rapid detection reagent for color development; 2) and comparing with a standard colorimetric card or a drawn standard curve to obtain the nitrate content of the water sample to be detected. The nitrate rapid detection reagent has strong adaptability and good stability, and can be matched with a standard colorimetric card or a portable colorimeter to carry out on-site rapid determination on nitrate in water quality and food samples.
Description
Technical Field
The invention relates to a rapid detection reagent and a detection method for nitrate.
Background
Nitrates and nitrites are the most common nitrogen-containing compounds in nature. Nitrite has similar appearance and taste to common salt, and can be used as food additive, but excessive nitrite in food can cause poisoning of eaters. Nitrate in the human body can be converted into nitrite under the action of microorganisms, so that not only the content of nitrite but also the content of nitrate needs to be measured to determine whether water quality and food are safe. For example: white sugar is a common food additive, the content of nitrite and nitrate in white sugar prepared from different raw materials is greatly different, and the content of nitrate in general beet sugar is obviously higher than that of sucrose, so enterprises for producing beet sugar need to control the content of nitrite and nitrate in finished products.
The method for measuring nitrate in the laboratory mainly comprises an ultraviolet photometry, a chromium-acid photometry, a cadmium column reduction-diazo coupling color photometry and the like. The ultraviolet photometry requires a large-scale spectrophotometer and is greatly interfered by a substrate. The chromaking photometry needs concentrated sulfuric acid, and the reagent is easy to change color and is not easy to store. The cadmium column reduction-diazo coupling color development photometry is complex and time-consuming in operation, and the cadmium column is aged along with the increase of the use times, so that the test data are unstable. At present, nitrate test paper method and cadmium powder test method are mainly used as the rapid detection method of nitrate. The sensitivity and the test accuracy of the nitrate test paper method are low, and the nitrate test paper method is not suitable for high-precision test. The existing cadmium powder testing method has the problem of poor repeatability, and the test results may have fold difference when the same sample is tested for multiple times.
Therefore, it is necessary to develop a rapid nitrate detection reagent and a rapid nitrate detection method that have high sensitivity, simple operation, and high measurement accuracy.
Disclosure of Invention
The invention aims to provide a rapid detection reagent and a detection method for nitrate.
The technical scheme adopted by the invention is as follows:
a rapid nitrate detection reagent comprises a reagent I and a reagent II, wherein:
the reagent I comprises the following components in percentage by mass: organic acid: 58% -88.5%, cadmium powder: 1% -10%, sulfanilamide: 0.5% -2%, polyvinylpyrrolidone: 10% -30%;
the reagent II comprises the following components in percentage by mass: naphthyl ethylenediamine hydrochloride: 2% -6%, organic acid: 94 to 98 percent.
Preferably, the organic acid is one of tartaric acid and citric acid.
Preferably, the particle size of the cadmium powder is less than 38 μm.
Preferably, the rapid nitrate detection reagent comprises a reagent I and a reagent II, wherein:
the reagent I comprises the following components in percentage by mass: tartaric acid: 75.7% and cadmium powder: 3.3%, sulfanilamide: 1%, polyvinylpyrrolidone: 20 percent;
the reagent II comprises the following components in percentage by mass: naphthyl ethylenediamine hydrochloride: 2%, tartaric acid: 98 percent.
Preferably, the reagent I and the reagent II are independently packaged, the mass of each part of the reagent I is 0.2-0.3 g, and the mass of each part of the reagent II is 0.1-0.2 g.
A method for carrying out on-site rapid detection on nitrate by adopting the rapid detection reagent for nitrate comprises the following steps:
1) adding a water sample to be tested into a clean colorimetric bottle, adding a reagent I, shaking for 2-4 min, standing for 2-4 min, adding a reagent II, shaking uniformly, and standing for 10-15 min;
2) and comparing colors of the standard colorimetric card to obtain the nitrate content of the water sample to be detected, or drawing a standard curve and detecting by using a portable colorimeter or a spectrophotometer to obtain the nitrate content of the water sample to be detected.
Preferably, the shaking mode in the step 1) is manual shaking or vortex instrument shaking, and the shaking frequency is more than 250 times/min.
Preferably, the method for manufacturing the standard color chart in step 2) is as follows:
1) preparing nitrate nitrogen standard solutions with the concentrations of 0.05mg/L, 0.1mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L, 0.5mg/L, 0.7mg/L and 1.0mg/L by using pure water;
2) adding a nitrate nitrogen standard solution with the concentration of 0.05mg/L into a clean colorimetric bottle, adding a reagent I, shaking for 2-4 min, standing for 2-4 min, adding a reagent II, shaking uniformly, standing for 10-15 min, observing the color of the solution, finding out a corresponding standard color on a pantone color card, repeating the operation, and testing the nitrate nitrogen standard solutions with the concentrations of 0.1mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L, 0.5mg/L, 0.7mg/L and 1.0mg/L respectively;
3) and (5) comparing the pantone color card to obtain the color value of each standard color, and carrying out color matching and printing by a computer to obtain the standard color comparison card.
The invention has the beneficial effects that: the nitrate rapid detection reagent has strong adaptability and good stability, and can be matched with a standard colorimetric card or a portable colorimeter to carry out on-site rapid determination on nitrate in water quality and food samples.
1) The rapid nitrate detection reagent disclosed by the invention is added with polyvinylpyrrolidone (PVP), and has the following effects in 4 aspects: the PVP can promote the fusion of the cadmium powder and other components, so that the separation and layering of the reagent in the processes of uniformly mixing and packaging can be avoided, and the distribution uniformity of the cadmium powder in the reagent is ensured; the PVP can remove static electricity, so that the problem that the cadmium powder is rubbed with a container to generate static electricity to be adsorbed on the wall of the container in the process of uniformly mixing and packaging the reagent is avoided, and the problem of inaccurate test result caused by loss of the cadmium powder is avoided; the PVP can enhance the dispersibility of the cadmium powder in the solution during reaction by utilizing the surface activity of the PVP, so that the problem of reduction efficiency reduction caused by the dissolution and the depolymerization of the cadmium powder in an acidic solution can be avoided; PVP is easily dissolved in water, and is chemically inert, so that the reduction and color development effects are not influenced;
2) the invention further controls the particle size of the cadmium powder below 38 mu m, thereby not only improving the uniformity of the particle size of the cadmium powder and ensuring the effective surface area to be consistent when the cadmium powder is reduced, but also further avoiding the cadmium powder from being separated and layered from other components;
3) the invention optimizes the shaking time, frequency and reaction time of each step, adopts the reagent I to carry out full reduction and diazotization reaction, and then adopts the reagent II to carry out coupling chromogenic reaction, thereby shortening the chromogenic reaction time, ensuring that the absorbance can quickly reach stability, and obviously improving the controllability of the reaction.
Detailed Description
The invention will be further explained and illustrated with reference to specific examples.
Example 1:
a rapid nitrate detection reagent comprises a reagent I and a reagent II, wherein:
the reagent I consists of the following components: tartaric acid: 75.70g, cadmium powder (particle size less than 38 μm): 3.30g, sulfanilamide: 1.00g, PVP: 20.00 g; fully and uniformly mixing all components in the reagent I, and packaging on an automatic powder packaging machine, wherein the specification is 0.30 g/bag;
the reagent II consists of the following components: naphthyl ethylenediamine hydrochloride: 2.00g, tartaric acid: 98.00 g; and fully and uniformly mixing all the components in the reagent II, and packaging on an automatic powder packaging machine, wherein the specification is 0.20 g/bag.
Example 2:
a rapid nitrate detection reagent comprises a reagent I and a reagent II, wherein:
the reagent I consists of the following components: tartaric acid: 74.00g, cadmium powder (particle size less than 38 μm): 5.00g, sulfanilamide: 1.00g, PVP: 20.00 g; fully and uniformly mixing all components in the reagent I, and packaging on an automatic powder packaging machine, wherein the specification is 0.20 g/bag;
the reagent II consists of the following components: naphthyl ethylenediamine hydrochloride: 4.00g, tartaric acid: 96.00 g; and fully and uniformly mixing all the components in the reagent II, and packaging on an automatic powder packaging machine, wherein the specification is 0.10 g/bag.
Example 3:
the method for manufacturing the standard colorimetric card comprises the following steps:
1) preparing nitrate nitrogen (NO as NO) with concentration of 0.05mg/L, 0.1mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L, 0.5mg/L, 0.7mg/L, 1.0mg/L with pure water3-N counts) standard solutions;
2) adding a nitrate nitrogen standard solution with the concentration of 0.05mg/L into a clean colorimetric bottle to a scale mark of 10mL, adding 1 package of the reagent I prepared in the embodiment 1, shaking for 3min on a vortex instrument, standing for 2min, adding 1 package of the reagent II prepared in the embodiment 1, shaking for 5min on the vortex instrument, standing for 10min, placing the colorimetric bottle on a white PVC plate, observing the color of the solution from the bottle opening downwards, finding out a corresponding standard color on a Pantong color card, repeating the operation, and testing the nitrate nitrogen standard solutions with the concentrations of 0.1mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L, 0.5mg/L, 0.7mg/L and 1.0mg/L respectively;
3) and (5) comparing the pantone color card to obtain the color value of each standard color, and carrying out color matching and printing by a computer to obtain the standard color comparison card.
Test example 1:
with reference to the example 1 of the present invention,the content of PVP in the reagent I is respectively adjusted to be 0, 5.0%, 10.0%, 20.0% and 30.0%, and the content of tartaric acid is correspondingly adjusted to be: 95.7 percent, 90.7 percent, 85.7 percent, 75.7 percent and 65.7 percent of the rest components are unchanged in content and are respectively prepared into nitrate rapid detection reagents, and nitrate nitrogen (with NO) with the concentration of 1.0mg/L is adopted3-N meter) standard solution, oscillating for 3min at high speed on a vortex instrument, performing standard chromogenic solution absorbance value test on a spectrophotometer at the wavelength of 525nm by using pure water as a reference and adopting a 1cm cuvette after color development, and sampling each reagent for testing at different time periods of packaging (total testing is 5 times), wherein the test results are shown in the following table:
TABLE 1 Effect of PVP content on test results
As can be seen from Table 1:
1) the content of PVP has great influence on the test effect;
2) when the formula does not contain PVP, the color development sensitivity is not high, the test repeatability is not good (RSD is 7.9%), loss caused by that part of the cadmium powder is adsorbed on the container wall and an aluminum foil packaging bag is observed by visual observation, the cadmium powder has an obvious caking phenomenon after reaction, and the cadmium powder is separated and layered in the processes of uniformly mixing and packaging;
3) with the increase of the addition of PVP, the test sensitivity and the test precision are gradually improved, the cadmium powder agglomeration condition is observed by visual observation to gradually disappear, and the adsorption and separation layering phenomena in the processes of uniformly mixing and packaging do not occur any more;
4) in order to ensure the sensitivity and the precision of the test, the content of PVP in the reagent I must be more than or equal to 10%, but the content of PVP is not more than 30% (the PVP content is too high and is easy to absorb moisture so that smooth blanking can not be realized during packaging), and the optimal content is 20%.
Test example 2:
commercially available cadmium powder (purity: 5N) was sieved out with 80, 150, 200, 325, 400, and 500 mesh standard sieves, respectively, and then nitrate rapid test reagents were prepared according to example 1,nitrate nitrogen (in NO) was used at a concentration of 1.0mg/L3-N meter) standard solution, oscillating for 3min at high speed on a vortex instrument, performing standard chromogenic solution absorbance value test on a spectrophotometer at the wavelength of 525nm by using pure water as a reference and adopting a 1cm cuvette after color development, and performing 5 parallel tests on each group of reagents respectively, wherein the test results are shown in the following table:
TABLE 2 influence of cadmium powder particle size on test parallelism
As can be seen from Table 2: the particle size of the cadmium powder has a great influence on the test repeatability and the color development sensitivity. When the particle size of the cadmium powder is larger, the cadmium powder is difficult to be uniformly distributed in the reagent (obvious separation and layering phenomena can be seen visually, and all parts of the cadmium powder are gathered at the bottom); along with the reduction of the particle size of the cadmium powder, the cadmium powder is gradually and uniformly distributed in the reagent, the effective reduction surface area is increased, and the repeatability and the color development sensitivity of the test are gradually improved; to ensure the test effect, the mesh number of the cadmium powder must be larger than 400 mesh (i.e., the particle size is smaller than 38 μm).
Test example 3:
the rapid detection reagent for nitrate prepared in example 1 was used, nitrate nitrogen (in NO) was used at a concentration of 1.0mg/L3-N meter) standard solution, oscillating at high speed on a vortex meter, changing the shaking time (the shaking time after reagent I is added in step 1 of the nitrate field rapid detection method), and performing the test of the absorbance value of the standard developing solution on a spectrophotometer at the wavelength of 525nm by using pure water as a reference and a 1cm cuvette after different developing time, wherein the test results are shown in the following table:
TABLE 3 Effect of shaking time
As can be seen from Table 3: the shaking time has great influence on the test result, the color development absorbance is improved along with the prolonging of the shaking time when the shaking time is less than 3min, the color development stabilization time is gradually shortened to 5min, the test result is reduced after the shaking time is more than 3min, the nitrate nitrogen is possibly excessively reduced to ammonia nitrogen, and the vortex shaking for 3min is selected to be most suitable.
Test example 4:
and testing by adopting an actual water sample, a white sugar sample and a syrup sample.
1) Sample pretreatment:
white sugar sample (provided by the research institute of Chinese food): 5.00g of white sugar is weighed and prepared into 100mL of sample solution by deionized water or pure water. Test results multiplied by dilution factor: 20.
syrup sample (provided by the center research institute): weigh 1.00g of syrup and prepare 1000mL of sample solution with deionized water or pure water. Test results multiplied by dilution factor: 1000 (syrup sample is dark brown, and the influence caused by the background color can be just eliminated by diluting more than 1000 times).
2) Drawing a standard curve:
nitrate standard solutions with the concentration (calculated by N) of 0.1mg/L, 0.2mg/L, 0.4mg/L, 0.6mg/L, 0.8mg/L and 1.0mg/L are prepared. Taking 1 clean colorimetric bottle, adding a standard solution to a scale mark of 10mL, adding 1 package of the reagent I prepared in the example 1, shaking at a high speed for 3min on a vortex instrument, standing for 2min, adding 1 package of the reagent II prepared in the example 1, shaking uniformly for dissolving, standing for reaction for 10min, performing a standard color development solution absorbance value test on a spectrophotometer at a wavelength of 525nm by using pure water as a reference and adopting a 1cm cuvette, and drawing a standard curve.
The standard curve equation is: C-0.4679A-0.0069, R20.9999 (the equation is very linear and the test range is 0-1.0 mg/L), wherein C is the nitrate concentration and A is the color development absorbance.
3) Testing of the sample:
taking a clean colorimetric bottle, adding a water sample to 10mL of scale marks, developing according to a standard solution test method, placing a colorimetric tube in a middle blank space of the standard colorimetric card prepared in the embodiment 3, carrying out visual colorimetry from a bottle opening downwards, wherein the color gradation indicating concentration which is the same as the color tone of the solution in the bottle is the content of nitrate in the sample solution, simultaneously carrying out absorbance test on a DR3900 spectrophotometer, and substituting the test into the standard curve equation to calculate the content of the nitrate. If the water sample test result exceeds the measuring range, the water sample can be properly diluted and then tested. And simultaneously, a standard recovery rate test is carried out according to a photometric test result. The water sample test results are shown in the following table:
table 4 water sample test results
Note: and (3) when the actual water sample is used for testing, the actual water sample exceeds the range of measurement, the actual water sample is diluted by 5 times and then is tested again, and the test result is obtained by multiplying the measured data by the dilution times. In the standard addition test, a standard substance is added to a raw water sample.
Taking a clean colorimetric bottle, adding a white sugar sample or a syrup sample to 10mL of scale marks, developing according to a standard solution test method, placing a colorimetric tube in a middle blank space of the standard colorimetric card prepared in the embodiment 3, carrying out visual colorimetry from a bottle opening downwards, wherein the color gradation indicating concentration which is the same as the color tone of the solution in the bottle is the content of nitrate in the sample solution, simultaneously carrying out absorbance test on a DR3900 spectrophotometer, and substituting the absorbance into the standard curve equation to calculate the content of nitrate. And simultaneously, a standard recovery rate test is carried out according to a photometric test result. The results of the white sugar and syrup tests are shown in the following table:
TABLE 5 white sugar and syrup samples test results
Note: in order to facilitate the standard adding test, the standard substance is directly added into the diluent, and the result is the direct test result of the diluent and is not multiplied by the dilution factor.
From tables 4 and 5, it can be seen that: the test results of the colorimetric card method and the photometric method are relatively consistent, and the method is very suitable for field application. The method is high in test sensitivity, is particularly suitable for dark samples such as syrup and the like, and can still accurately test results after the samples are diluted to be almost colorless. The standard adding recovery rate of the test is between 96.00% and 102.00%, which indicates that the river water sample, the white sugar sample and the syrup sample can be directly measured after being simply prepared or diluted, and the test result is reliable.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (5)
1. A nitrate rapid detection reagent is characterized in that: comprising a reagent I and a reagent II, wherein:
the reagent I comprises the following components in percentage by mass: tartaric acid: 58% -88.5%, cadmium powder: 1% -10%, sulfanilamide: 0.5% -2%, polyvinylpyrrolidone: 10% -30%;
the reagent II comprises the following components in percentage by mass: naphthyl ethylenediamine hydrochloride: 2% -6%, tartaric acid: 94% -98%;
the particle size of the cadmium powder is less than 38 micrometers;
and independently subpackaging the reagent I and the reagent II, wherein the mass of each part of the reagent I is 0.2-0.3 g, and the mass of each part of the reagent II is 0.1-0.2 g.
2. The reagent for rapid detection of nitrate according to claim 1, which is characterized in that: comprising a reagent I and a reagent II, wherein:
the reagent I comprises the following components in percentage by mass: tartaric acid: 75.7% and cadmium powder: 3.3%, sulfanilamide: 1%, polyvinylpyrrolidone: 20 percent;
the reagent II comprises the following components in percentage by mass: naphthyl ethylenediamine hydrochloride: 2%, tartaric acid: 98 percent.
3. A nitrate on-site rapid detection method is characterized in that: the rapid nitrate detection reagent as claimed in any one of claims 1 to 2 is used for detection, and specifically comprises the following steps:
1) adding a water sample to be tested into a clean colorimetric bottle, adding a reagent I, shaking for 2-4 min, standing for 2-4 min,
adding the reagent II, shaking up, and standing for 10-15 min;
2) and comparing colors of the standard colorimetric card to obtain the nitrate content of the water sample to be detected, or drawing a standard curve and detecting by using a portable colorimeter or a spectrophotometer to obtain the nitrate content of the water sample to be detected.
4. The on-site rapid detection method for nitrate according to claim 3, characterized in that: the shaking mode of the step 1) is that the vortex instrument shakes, and the shaking frequency is more than 250 times/min.
5. The on-site rapid detection method for nitrate according to claim 3, characterized in that: the manufacturing method of the standard colorimetric card in the step 2) is as follows:
1) preparing nitrate nitrogen standard solutions with the concentrations of 0.05mg/L, 0.1mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L, 0.5mg/L, 0.7mg/L and 1.0mg/L by using pure water;
2) adding a nitrate nitrogen standard solution with the concentration of 0.05mg/L into a clean colorimetric bottle, adding a reagent I, shaking for 2-4 min, standing for 2-4 min, adding a reagent II, shaking uniformly, standing for 10-15 min, observing the color of the solution, finding out a corresponding standard color on a pantone color card, repeating the operation, and testing the nitrate nitrogen standard solutions with the concentrations of 0.1mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L, 0.5mg/L, 0.7mg/L and 1.0mg/L respectively;
and (5) comparing the pantone color card to obtain the color value of each standard color, and carrying out color matching and printing by a computer to obtain the standard color comparison card.
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| CN111610183A (en) * | 2020-05-26 | 2020-09-01 | 王淑娟 | Semi-quantitative detection method for total residual chlorine in wastewater |
| CN111610184A (en) * | 2020-05-26 | 2020-09-01 | 王淑娟 | Semi-quantitative detection method for total residual chlorine in water |
| CN113418913A (en) * | 2021-06-24 | 2021-09-21 | 神美科技有限公司 | Stable nitrite test paper and preparation method thereof |
| CN113670902A (en) * | 2021-07-02 | 2021-11-19 | 四川师范大学 | Detection agent for rapidly and quantitatively determining nitrite and detection method thereof |
| CN115436585A (en) * | 2022-07-25 | 2022-12-06 | 广州伊创科技股份有限公司 | Online intelligent labeling method for water quality monitoring |
| CN115575390B (en) * | 2022-09-30 | 2023-05-30 | 广东省环境科学研究院 | Combined reagent for monitoring water quality and application method thereof |
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