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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，以酶标仪测定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。
| N1510410 | Component | 100T | Storage |
| N1510410A | 硝态氮标准 (200μg/mL) | 1 mL | 2-8℃. |
| N1510410B | 硝态氮提取液 | 500 mL | RT. |
| N1510410C | Assay Buffer | 500 mL | RT. |
| N1510410D | 磺胺混合粉剂 | 10 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：
加入物（mL） | 1 | 2 | 3 | 4 | 5 | 6 |
硝态氮标准 (20μg/mL) | 0.05 | 0.1 | 0.2 | 0.3 | 0.4 | 0.5 |
蒸馏水 | 0.45 | 0.4 | 0.3 | 0.2 | 0.1 | 0 |
硝态氮浓度 (μg/mL) | 2 | 4 | 8 | 12 | 16 | 20 |
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，取上清液加入96孔板中，以空白管做对照调零，酶标仪520nm处测定标准孔、测定孔吸光度（记为A<sub>标准</sub>、A<sub>测定</sub>）。
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，根据测定孔的吸光-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、如果没有酶标仪，也可以使用普通的分光光度计测定。
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 within plant tissues reflects the supply of nitrate nitrogen in the soil and can serve as an indicator of soil nitrogen fertility. Determining the nitrate nitrogen content in plants is not only important for assessing plant nitrogen nutrition but also holds significant value for evaluating the quality of vegetables and processed products made from plant materials.
Detection Principle
A reducing agent converts nitrate into nitrite, which then reacts with sulfanilic acid and naphthylamine to form a rose-red azo compound. The color intensity is directly proportional to the nitrogen content within a certain range. The absorbance at 520 nm is measured using a microplate reader, 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 intended for research purposes only and is not suitable for clinical diagnosis or other applications.
| N1510410 | Component | 100T | Storage |
| N1510410A | Nitrate Nitrogen Standard (200 µg/mL) | 1 mL | 2-8℃. |
| N1510410B | Nitrate Nitrogen Extraction Solution | 500 mL | RT. |
| N1510410C | Assay Buffer | 500 mL | RT. |
| N1510410D | Sulfanilamide Mixture Powder | 10 g | RT. Store in the dark. |
Reagents, consumables and Equipments not provided
Procedure
1. Sample Preparation
1.1 Plant Samples
Take 0.1–0.15 g of fresh plant tissue (from normal or stressed conditions). Wash, dry, and mince into 1–2 mm pieces. Add 4 mL of Nitrate Nitrogen Extraction Solution. Vortex vigorously for 2–4 minutes. After clarification, the supernatant is the nitrate nitrogen extract. Store at 4°C for later use.
1.2 Serum and Urine Samples
Samples prepared by conventional methods can be used directly for assay with this kit. Store at 4°C for nitrate nitrogen detection.
1.3 High-Concentration Samples
If the sample contains a high concentration of nitrate nitrogen, dilute appropriately using the 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:
Reagent (mL) | 1 | 2 | 3 | 4 | 5 | 6 |
Nitrate Nitrogen Standard (20 µg/mL) | 0.05 | 0.1 | 0.2 | 0.3 | 0.4 | 0.5 |
Distilled water | 0.45 | 0.4 | 0.3 | 0.2 | 0.1 | 0 |
Nitrate Nitrogen Concentration (µg/mL) | 2 | 4 | 8 | 12 | 16 | 20 |
3. Loading
Set up blank tubes, standard tubes, and sample tubes according to the table below. Add solutions in the specified order, avoiding bubble formation. If the sample nitrate nitrogen concentration is too high, reduce the sample volume or dilute appropriately before assay. It is best to set up duplicate tubes for samples.
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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 96-well plate. Using the blank tube as a reference to zero the instrument, measure the absorbance of the standard wells and sample wells at 520 nm using the microplate reader (recorded as A<sub>standard</sub> and A<sub>sample</sub>).
5. Calculation
Using the concentration (µg/mL) of the nitrate nitrogen standard series (Tubes 1–6) as the x-axis and the corresponding absorbance as the y-axis, generate a standard curve. Calculate the nitrate nitrogen content of the sample based on the absorbance of the sample well.
For plant tissue samples:
Nitrate nitrogen (µg/g) = C × V / m
C: Nitrate nitrogen concentration of the sample obtained from the standard curve (µg/mL)
V: Total volume of the sample extract (mL); in this method, it is 4 mL
m: Sample weight (g)
For serum, urine, and other liquid samples:
Nitrate nitrogen (µg/mL) = C × N
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 assayed immediately after collection, store at 4°C.
The Assay Buffer is volatile; please store it sealed tightly.
If a microplate reader is unavailable, a standard spectrophotometer can also be used for measurement.
If the concentration of the sample is too high, dilute the sample with Nitrate Nitrogen Extraction Solution or distilled water and repeat the assay.
For your safety and health, please wear a lab coat and disposable gloves during operation.
After opening, use the reagent as soon as possible to avoid affecting subsequent experimental results,
| N1510410 | Component | 100T | Storage |
| N1510410A | 硝态氮标准 (200μg/mL) | 1 mL | 2-8℃. |
| N1510410B | 硝态氮提取液 | 500 mL | RT. |
| N1510410C | Assay Buffer | 500 mL | RT. |
| N1510410D | 磺胺混合粉剂 | 10 g | RT. Store in the dark. |
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0332898 |
分析证书 | N1510410 |
| ZJ26F0332897 |
分析证书 | N1510410 |
| ZJ26F0332896 |
分析证书 | N1510410 |
| ZJ26F0332895 |
分析证书 | N1510410 |
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