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Method for rapidly detecting ammonia nitrogen in water on large batches by microplate reader micro-colorimetric method

A microplate reader and colorimetric technology, applied in the field of ammonia nitrogen detection and analysis in water, can solve problems such as few application reports, and achieve the effect of reducing emissions and saving time in analysis and detection work.

Inactive Publication Date: 2019-07-16
长江水利委员会水文局
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AI Technical Summary

Problems solved by technology

The application of microplate reader is mainly in biomedical detection, and there are few reports on the application in the field of water quality detection

Method used

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  • Method for rapidly detecting ammonia nitrogen in water on large batches by microplate reader micro-colorimetric method
  • Method for rapidly detecting ammonia nitrogen in water on large batches by microplate reader micro-colorimetric method
  • Method for rapidly detecting ammonia nitrogen in water on large batches by microplate reader micro-colorimetric method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 standard series preparation

[0028] Spectrophotometric method Pipette 0.00, 0.50, 1.00, 2.00, 4.00, 6.00, 8.00 and 10.00mL ammonia nitrogen standard solution into a 50mL stoppered graduated tube, add water to 50mL. Assays were performed according to standard methods.

[0029] For the micro-colorimetric method of microplate reader, take 0, 2, 4, 8, 16, 24, 32 and 40 μL of ammonia nitrogen standard solution in a 96-well microplate, add water to 200 μL, and the corresponding ammonia nitrogen content is consistent with the spectrophotometric method. When making a standard curve with different sample volumes, pipette the ammonia nitrogen standard solution in proportion.

Embodiment 2

[0030] Embodiment 2 experimental method

[0031] Nessler's reagent spectrophotometric determination adopts the standard method (HJ 535-2009). Add 1.0mL 500g / L potassium sodium tartrate solution, shake well, then add Nessler's reagent 1.0mL, shake well. After standing at room temperature for 10 minutes, use a cuvette with an optical path of 20 mm, and use water as a reference at a wavelength of 420 nm to measure the absorbance.

[0032] Microplate reader microcolorimetric method corresponds to adding 4 μL potassium sodium tartrate solution and 4 μL Nessler’s reagent. After developing color under optimized conditions, measure the optical density (OD) at a wavelength of 420 nm. When making a standard curve with different sample volumes, pipette the sodium potassium tartrate solution and Nessler's reagent in proportion.

[0033] Use the linear range, detection limit, minimum detection limit, precision and sensitivity analysis parameters to optimize the results; the linear range ...

Embodiment 3

[0035] Example 3 Sample volume

[0036] The light source of the microplate reader is transmitted from the bottom of the microwell plate, and the sample volume should cover the bottom of the microwell without overflowing the microwell. The working volume of the Thermo 96-well microplate used in the experiment was 50-250 μL, and 5 sample volume gradients of 50, 100, 150, 200 and 250 μL were set. figure 1 It is the standard curve of the same concentration of ammonia nitrogen standard sequence samples under different sample volume gradients. The correlation coefficients of the curves drawn for different volume samples can reach 0.999. With the increase of sample amount, the optical density of the same concentration sample gradually increased, and the slope of the standard curve also gradually increased. The slope of the standard curve has a more obvious impact on the calculation of the detection value of the standard sample. See Table 1 for the measurement results of the standa...

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Abstract

The invention relates to a method for rapidly detecting ammonia nitrogen in water on large batches by a microplate reader micro-colorimetric method, and belongs to the technical field of detection analysis of ammonia nitrogen in water. A method for rapidly analyzing ammonia nitrogen in the water on large batches is established by the microplate reader micro-colorimetric method, optimum parametersand conditions of the method are determined, and the microplate reader micro-colorimetric method is compared with a spectrophotometric method. The microplate reader micro-colorimetric method and a traditional nessler reagent spectrophotometric method have no differences in analyzing parameters such as a linearity range, a detection limit, a lowest detection limit and sensitivity. Through researchon experiment conditions, only 200[mu]L of sample volume, 4[mu]L of a nessler reagent are needed to detect ammonia nitrogen by the microplate reader micro-colorimetric method, detection of 96 samplescan be finished in 1 min, deficiencies of the spectrophotometric method in batch operation are overcome, discharging of experiment wastewater is reduced, and requirements of rapid, precise and time-saving analysis detecting work are met.

Description

technical field [0001] The invention belongs to the technical field of detection and analysis of ammonia nitrogen in water, and in particular relates to a method for rapid detection of ammonia nitrogen in large batches by using micro-colorimetry with a microplate reader. Background technique [0002] Accelerate the reform of the ecological civilization system and build a beautiful new China. This move has accelerated a series of processes such as promoting green development, focusing on solving outstanding environmental problems, and increasing ecological system protection. Especially in the comprehensive promotion of the "river and lake chief" work , the monitoring and supervision of water pollutants has become the focus of governments at all levels. Ammonia nitrogen is one of the four binding indicators for the reduction of major pollutants in the country, and it is also one of the important indicators for evaluating the health of rivers, lakes and reservoirs. As a basic ...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/17G01N21/78
CPCG01N21/17G01N21/31G01N21/78
Inventor 钱宝徐剑秋张明波汪金成肖潇王文静陈峰张德兵周裕红李炜钦
Owner 长江水利委员会水文局
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