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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
硝态氮是植物最主要的氮源，植物体内硝态氮含量反映了土壤中硝态氮的供应情况，可作为土壤氮肥的指标，测定植物体内的硝态氮含量，不仅能够反映出植物的氮素营养情况，而且对鉴定蔬菜和以植物为原料的加工制品的品质也有重要的意义。
检测原理：还原剂将硝酸根还原成亚硝酸根后，与对氨基苯磺酸和萘胺结合，形成玫瑰红色的偶氮化合物，其颜色深浅与氮含量在一定范围内呈正比，以分光光度计测定520nm处吸光度，根据硝态氮的标准曲线即可计算出样品的硝态氮含量，主要用于测定植物组织、血清、组织样本等的硝态氮含量。该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。
| N1510390 | Component | 50T | Storage |
| N1510390A | 硝态氮标准 (200μg/mL) | 1 mL | 2-8℃. |
| N1510390B | 硝态氮提取液 | 250 mL | RT. |
| N1510390C | Assay Buffer | 250 mL | RT. |
| N1510390D | 磺胺混合粉剂 | 5 g | RT. Store in the dark. |
自备仪器和试剂
操作步骤
1、准备样品
1.1 植物样品
取正常或逆境下的新鲜植物组织0.1~0.15g，清洗干净，擦干，剪碎成1~2mm的碎片，加入4ml硝态氮提取液，剧烈振荡2~4min，静置澄清后，上清液即为硝态氮提取液，4℃保存备用。
1.2 血清和尿液样品
按照常规方法制备后可以直接用于该试剂盒的测定，4℃保存，用于硝态氮的检测。
1.3 高活性样品
如果样品中含有较高浓度的硝态氮，可以使用硝态氮提取液或蒸馏水进行恰当的稀释。
2、配制系列硝态氮标准溶液
取硝态氮标准 (200μg/mL) 用蒸馏水稀释至20μg/mL，再按下表继续稀释：
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3、加样
按照下表设置空白管、标准管、测定管，溶液应按照顺序依次加入，并注意避免产生气泡。如果样品中的硝态氮浓度过高，可以减少样品用量或适当稀释后再进行测定，样品的检测最好能设置平行管。
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4、硝态氮测定
剧烈振荡1min，静置10min，用双层滤纸过滤至新的离心管中，4000rpm离心5min，取上清液再次离心，上清液加入光径为1cm的比色皿中，以空白管调零，分光光度计520nm处测定标准管、测定管吸光度（记为A标准、A测定）。
5、结果计算
以系列硝态氮标准 (1~6号管) 浓度 (μg/mL) 为横坐标，以对应的吸光度为纵坐标，制作标准曲线，根据测定管的吸光度计算出样品硝态氮的含量。根据如下公式计算具体样品中硝态氮的含量：
植物组织样品硝态氮 (μg/g)=C×V/m
参数说明：
C：从标准曲线上查得的样品的硝态氮浓度 (μg/mL)
V：样品提取液的总体积 (ml)，本法中为4mL
m：样品的质量(g)
血清、尿液等样品硝态氮(μg/mL)=C×N
参数说明：
C：从标准曲线上查得的样品的硝态氮浓度 (μg/mL)
N：稀释倍数
注意事项
1、实验材料应尽量新鲜，如取材后不立即测定，应存于4℃。
2、Assay Buffer易挥发，请密闭保存。
3、如果没有分光光度计，也可以使用普通的酶标仪测定。
4、所测样品的浓度过高时，应用硝态氮提取液或蒸馏水稀释样品后重新测定。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
6、试剂开封后请尽快使用，以防影响后续实验效果。
Nitrate nitrogen is the primary nitrogen source for plants. The nitrate nitrogen content in plant tissues reflects the supply of nitrate nitrogen in the soil and can serve as an indicator of soil nitrogen fertility. Measuring the nitrate nitrogen content in plants not only reveals the nitrogen nutrition status of the plants but is also significant for assessing the quality of vegetables and processed products derived from plants.
Detection Principle
Nitrate is reduced to nitrite by a reducing agent. The nitrite then reacts with sulfanilic acid and naphthylamine to form a rose-red azo compound. The intensity of the color is proportional to the nitrogen content within a certain range. The absorbance is measured at 520 nm using a spectrophotometer, and the nitrate nitrogen content of the sample is calculated based on a standard curve. This kit is primarily used to determine nitrate nitrogen content in plant tissues, serum, tissue samples, etc. It is for research use only and is not suitable for clinical diagnosis or other purposes.
| N1510390 | Component | 50T | Storage |
| N1510390A | Nitrate Nitrogen Standard (200 µg/mL) | 1 mL | 2-8℃. |
| N1510390B | Nitrate Nitrogen Extraction Solution | 250 mL | RT. |
| N1510390C | Assay Buffer | 250 mL | RT. |
| N1510390D | Sulfanilamide Mixture Powder | 5 g | RT. Store in the dark. |
Reagents, consumables and Equipments not provided
Operating Steps
1. Sample Preparation
1.1 Plant Samples
Take 0.1–0.15 g of fresh plant tissue (normal or under stress), wash, dry, and cut into 1–2 mm pieces. Add 4 mL of Nitrate Nitrogen Extraction Solution, vortex vigorously for 2–4 minutes, and allow to stand until clear. The supernatant is the nitrate nitrogen extract. Store at 4°C for use.
1.2 Serum and Urine Samples
After preparation by conventional methods, they can be directly used for assay with this kit. Store at 4°C for nitrate nitrogen detection.
1.3 High-Activity Samples
If the sample contains a high concentration of nitrate nitrogen, dilute appropriately with Nitrate Nitrogen Extraction Solution or distilled water.
2. Preparation of Nitrate Nitrogen Standard Series
Dilute the Nitrate Nitrogen Standard (200 µg/mL) with distilled water to 20 µg/mL. Then continue diluting according to the table below:
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3. Sample Addition
Set up blank, standard, and test tubes according to the table below. Add solutions in the specified order, taking care to avoid bubbles. If the nitrate nitrogen concentration in the sample is too high, reduce the sample volume or dilute appropriately before measurement. It is recommended to set up duplicate tubes for sample detection.
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4. Nitrate Nitrogen Measurement
Vortex vigorously for 1 minute and let stand for 10 minutes. Filter through double-layer filter paper into a new centrifuge tube. Centrifuge at 4000 rpm for 5 minutes, take the supernatant and centrifuge again. Transfer the supernatant to a 1 cm pathlength cuvette. Zero the spectrophotometer with the blank tube and measure the absorbance of the standard and test tubes at 520 nm (recorded as A<sub>Standard</sub> and A<sub>Test</sub>).
5. Result Calculation
Using the nitrate nitrogen standard series (Tubes 1–6) concentrations (µg/mL) as the x-axis and the corresponding absorbance values as the y-axis, plot a standard curve. Calculate the nitrate nitrogen content of the sample based on the absorbance of the test tube. Use the following formulas:
For plant tissue samples:
Nitrate Nitrogen (µg/g) = C × V / m
Parameter Explanation:
C: Nitrate nitrogen concentration of the sample obtained from the standard curve (µg/mL)
V: Total volume of the sample extract (mL); 4 mL in this method.
m: Sample mass (g)
For serum, urine, and other liquid samples:
Nitrate Nitrogen (µg/mL) = C × N
Parameter Explanation:
C: Nitrate nitrogen concentration of the sample obtained from the standard curve (µg/mL)
N: Dilution factor
Precautions
Experimental materials should be as fresh as possible. If not measured immediately after collection, store at 4°C.
Assay Buffer is volatile; store tightly sealed.
If a spectrophotometer is unavailable, a standard microplate reader can also be used.
If the measured sample concentration is too high, dilute the sample with Nitrate Nitrogen Extraction Solution or distilled water and repeat the measurement.
For your safety and health, please wear a lab coat and disposable gloves during operation.
Please use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0434282 |
分析证书 | N1510390 |
危险品化学品经营许可证（带存储）