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Cité dans 0 publications à comité de lecture couvrant la chromatographie, la synthèse organique et les réactions de couplage croisé.
Nitrogen is the nutrient element with the highest demand and contribution to yield increase in crop growth. Nitrate nitrogen in the soil refers to the nitrogen element contained in nitrate salts. As one of the main forms of nitrogen absorbed by higher plants, its content is directly related to crop yield and quality. It can reflect the short-term nitrogen supply status of the soil and is significant for controlling nitrogen nutrition in the soil.
Assay Principle
NO₃⁻ can react with salicylic acid under concentrated acid conditions to generate nitrosalicylic acid, which appears yellow under alkaline conditions. The product has a characteristic absorption peak at 410 nm. The nitrate nitrogen content can be quantitatively detected by the change in absorbance.
| S1505722 | Component | 48T | 96T | Storage |
| S1505722A | Reagent Ⅰ | 1EA | 2EA | 2-8℃. Store in the dark. |
| S1505722B | Reagent Ⅱ | 28.5 mL | 57 mL | 2-8℃ |
| S1505722C | Standard | 1EA | 1EA | 2-8℃ |
Note: It is recommended to perform a pilot experiment with 2-3 samples expected to have significant differences before formal testing.
Required Materials Not Provided
1. Microplate reader or visible spectrophotometer (capable of measuring absorbance at 410 nm)
2. 96-well plate or micro glass cuvettes, adjustable pipettes and tips
3. Analytical balance, constant temperature water bath, incubator shaker, benchtop centrifuge
4. Deionized water, Concentrated Sulfuric Acid
Experimental Procedure
1. Reagent Preparation
| Reagent Name | Preparation Steps | Notes & Storage |
| Reagent Ⅰ | Reconstitute before use: Add 1.2 mL of concentrated sulfuric acid per vial. Dissolve thoroughly. | Stable at 4°C, protected from light, for 1 week. Highly corrosive – use appropriate personal protective equipment (PPE). |
| Reagent Ⅱ | Ready-to-use; equilibrate to room temperature before use. | Store at 4°C. Highly corrosive – use appropriate PPE. |
| Standard | Reconstitute before use: Add 1 mL deionized water to the standard vial to prepare a 10 mg/mL NO₃⁻ stock solution. Take 100 μL of this stock and add 900 μL deionized water to make a 1 mg/mL NO₃⁻ working standard solution. | Unused 10 mg/mL stock can be stored at 4°C for 2 weeks. |
2. Sample Preparation
Add deionized water to the soil sample at a ratio of 1:5-10 (soil weight (g) : water volume (mL)). (Adding 1 mL deionized water to approx. 0.1 g soil is recommended).
Extract by shaking at 37°C for 1 hour.
Centrifuge at 10,000 g for 10 minutes at 25°C.
Collect the supernatant for assay.
Note: Air-drying or oven-drying soil can easily alter nitrate nitrogen content. Using fresh soil or samples processed identically is recommended.
If not used immediately, samples can be stored at -80°C for several weeks. Control thawing temperature and time during assay. If thawing at room temperature, complete within 4 hours.
3. Assay Procedure
3.1 Instrument Preparation
Preheat the microplate reader or visible spectrophotometer for 30 minutes. Set wavelength to 410 nm.
Zero the visible spectrophotometer with deionized water.
3.2 Assay Setup (Perform in 1.5 mL centrifuge tubes)
| Reagent (μL) | Blank Tube | Standard Tube | Test Tube |
| Sample Supernatant | 0 | 0 | 10 |
| Deionized Water | 10 | 0 | 0 |
| Standard (1 mg/mL) | 0 | 10 | 0 |
| Reagent Ⅰ | 20 | 20 | 20 |
Mix thoroughly and let stand at 25°C for 30 minutes.
| Reagent Ⅱ | 475 | 475 | 475 |
3.3 Measurement
Mix well and vortex to ensure any precipitate formed is fully dissolved.
Transfer 200 μL to a micro glass cuvette or 96-well plate.
Measure the absorbance at 410 nm.
Record the values as ABlank, AStandard, and ATest. Calculate ΔATest = ATest - ABlank and ΔAStandard = AStandard - ABlank. Notes:
Notes:
(1) The Blank and Standard tubes only need to be measured once. A pilot experiment with 2-3 variable samples is recommended.
(2) If ΔATest is less than 0.002, consider increasing the sample amount. If ΔATest is greater than 1.5, dilute the sample supernatant further with deionized water and multiply the result by the dilution factor, or reduce the soil amount used for extraction.
(3) Soil colloids do not adsorb nitrate ions, which are easily soluble in water and mobile within the soil. When measuring multiple samples or replicates, ensure consistent sampling depth (recommended: approx. 10 cm below the surface). Note that content varies by region.
4. Calculation of Results
Note: The derived and simplified formulas provided are equivalent. The simplified formula (in bold) is recommended.
NO₃⁻ Content (μg/g sample) = ΔA test / (ΔA standard / C standard ) × Vtotal extract / W=1000 × ΔATest / ΔAStandard / W
Parameter Definitions:
W: Sample mass (g)
CStandard: Standard solution concentration (1,000 μg/mL)
Vtotal extract: Total extraction volume (1 mL)
5. Results Example
Soil: 0.1 g extracted with 1 mL deionized water for 1h, centrifuged.
Measured: ATest=0.062, ABlank=0.047, AStandard=1.515
ΔATest = 0.062 - 0.047 = 0.015
ΔAStandard = 1.515 - 0.047 = 1.468
NO₃⁻ Content (μg/g soil) = 1,000 × 0.015 / 1.468 / 0.1 = 102.18 μg/g soil
Precautions
1. It is recommended to perform a pilot experiment with 2-3 samples expected to have significant differences before formal testing.
2. This product is for research use only. Not for use in diagnostic procedures. For your safety and health, please wear lab coats and disposable gloves during operation.
Document complet couvrant les dangers, la manipulation, le stockage et la conformité réglementaire.
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| Lot Number | Certificate Type | Date | Article |
|---|---|---|---|
 ZJ26F0737529 | Certificate of Analysis | Jul 23, 2026 | S1505722 |
| ZJ26F0737530 | Certificate of Analysis | Jul 23, 2026 | S1505722 |
| Sensibilité | Light-sensitive |
|---|
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