BioReagent 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
2-8°C储存,避光,室温。低温运输 。请查阅批次 COA 获取详细规格。
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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。
磷是植物必需的大量元素。植物主要通过根系从土壤中获取磷元素。土壤中的磷主要以有机磷和无机磷的形式存在。而有机磷只有经过矿化分解转化为无机磷，才能进一步被植物吸收利用。
检测原理：从土壤中提取无机磷，在酸性环境中，通过钼蓝法定磷，测定在 660 nm 的吸光度，计算土壤中无机磷的含量。
适用样本：土壤样本
| S1506787 | Component | 48T | 96T | Storage |
| S1506787A | Extraction Buffer | 60 mL | 120 mL | 2-8℃ |
| S1506787B | Reagent Ⅰ | 1 EA | 1 EA | 2-8℃. Store in the dark. |
| S1506787C | Reagent Ⅱ | 1 EA | 1 EA | 2-8℃. Store in the dark. |
| S1506787D | Reagent Ⅲ | 2.5 mL | 5 mL | RT |
| S1506787E | Standard | 1 mL | 1 mL | 2-8℃ |
注意：正式检测前，建议选择 2-3 个预期差异较大的样本进行预实验。
自备仪器和试剂
1、酶标仪或可见分光光度计（能测 660 nm 处的吸光度）
2、96孔板或微量玻璃比色皿、可调节式移液枪及枪头
3、分析天平、可调式水浴锅、台式离心机、100 目筛网
4、去离子水
实验流程
1、试剂准备
| 试剂名称 | 试剂准备 | 注意事项 |
| Extraction Buffer | 即用型；使用前平衡到室温 | 4℃保存 |
| Reagent Ⅰ | 临用前配制，48T 加入2.5mL去离子水充分溶解，96T加入5mL去离子水充分溶解后使用 | 4℃避光保存可稳定1个月 |
| Reagent Ⅱ | 临用前配制，48T加入2.5mL去离子水充分溶解，96T加入5mL去离子水充分溶解后使用 | 4℃避光保存可稳定1个月 |
| Reagent Ⅲ | 即用型 | 室温保存 |
| Standard | 即用型；使用前平衡到室温 | 4℃保存 |
| 定磷剂 | 按照去离子水：Reagent Ⅰ：Reagent Ⅱ：Reagent Ⅲ=2:1:1:1 的比例配制，配好的定磷试剂应为浅黄色 | 若无色则试剂失效，若是蓝色则为磷污染（请根据需要，用多少配多少） |
2、样本制备
土壤样品：取适量风干土壤样品，去除石块、树枝等杂质，过100目筛网，准确称取0.01g，加入1mL Extraction Buffer，室温震荡混匀，然后置于40℃水浴浸提1h，8,000 g，25℃，离心10min，取上清液，待测。
3、实验步骤
3.1 酶标仪或可见分光光度计预热30min，调节波长到660nm。可见分光光度计用去离子水调零。
3.2 操作表（下述操作在1.5mL离心管中进行）：
| 试剂（μL） | 空白管 | 标准管 | 测定管 |
| 样本上清 | 0 | 0 | 40 |
| 去离子水 | 40 | 0 | 0 |
| 定磷剂 | 200 | 200 | 200 |
| Standard | 0 | 40 | 0 |
3.3 混匀后置于40℃水浴保温10min，室温冷却10min后取200μL加入96孔板/微量玻璃比色皿中于660nm测定吸光度，记为A空白、A标准、A测定，计算ΔA测=A测定-A空白、ΔA标=A标准-A空白。注意：空白孔和标准孔只需测定1次。实验之前建议选择2-3个预期差异大的样本做预实验。如果ΔA测小于0.01，可适当加大样本量。如果ΔA测大于1.0，样本可用Extraction Buffer进一步稀释，计算结果乘以稀释倍数，或减少提取用样本量。
4、结果计算
土壤样本无机磷含量计算
使用96孔板/微量玻璃比色皿测定的计算公式如下:
图1：试剂盒测定土壤中无机磷酸盐的含量
注意事项
1、定磷剂需临用前配制，限当天使用。
2、40min内完成检测。
3、本产品仅供科学研究使用，不适用于临床诊断。为了您的安全和健康，请穿实验服并戴一次性手套操作。
Phosphorus is an essential macronutrient for plants. Plants primarily absorb phosphorus from the soil through their root systems. Soil phosphorus exists mainly in organic and inorganic forms. Organic phosphorus must be mineralized and converted into inorganic phosphorus before it can be absorbed and utilized by plants.
Assay Principle:Inorganic phosphorus is extracted from the soil. In an acidic environment, phosphorus is determined by the molybdenum blue method. The absorbance at 660 nm is measured to calculate the inorganic phosphorus content in the soil.
Applicable Samples: Soil samples
| S1506787 | Component | 48T | 96T | Storage |
| S1506787A | Extraction Buffer | 60 mL | 120 mL | 2-8℃ |
| S1506787B | Reagent Ⅰ | 1 EA | 1 EA | 2-8℃. Store in the dark. |
| S1506787C | Reagent Ⅱ | 1 EA | 1 EA | 2-8℃. Store in the dark. |
| S1506787D | Reagent Ⅲ | 2.5 mL | 5 mL | RT |
| S1506787E | Standard | 1 mL | 1 mL | 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 660 nm)
2. 96-well plate or micro glass cuvettes, Adjustable pipettes and tips
3. Analytical balance, Adjustable water bath, Benchtop centrifuge, 100-mesh sieve
4. Deionized water
Experimental Procedure
1. Reagent Preparation
| Reagent Name | Preparation Steps | Notes & Storage |
| Extraction Buffer | Ready-to-use; equilibrate to room temperature before use. | Store at 4°C. |
| Reagent Ⅰ | Before use, dissolve the vial contents in 2.5 mL (48T) or 5 mL (96T) deionized water. Mix thoroughly. | Stable at 4°C protected from light for 1 month. |
| Reagent Ⅱ | Before use, dissolve the vial contents in 2.5 mL (48T) or 5 mL (96T) deionized water. Mix thoroughly. | Stable at 4°C protected from light for 1 month. |
| Reagent Ⅲ | Ready-to-use. | Store at Room Temperature. |
| Standard | Ready-to-use; equilibrate to room temperature before use. | Store at 4°C. |
| Phosphorus Assay Reagent | Prepare by mixing Deionized Water : Reagent I : Reagent II : Reagent III at a ratio of 2 : 1 : 1 : 1. The prepared reagent should be light yellow. | Prepare fresh before use. Discard if colorless (reagent degraded) or blue (phosphorus contaminated). Prepare as needed. |
2. Sample Preparation
Soil Sample: Take an appropriate amount of air-dried soil, remove impurities like stones and twigs, and pass through a 100-mesh sieve.
Accurately weigh 0.01 g of the sieved soil.
Add 1 mL of Extraction Buffer.
Shake or vortex to mix thoroughly at room temperature.
Incubate in a 40°C water bath for 1 hour for extraction.
Centrifuge at 8,000 g, 25°C for 10 minutes.
Collect the supernatant for assay.
3. Assay Procedure
3.1 Instrument Preparation
Preheat the microplate reader or visible spectrophotometer for 30 minutes. Set wavelength to 660 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 | 40 |
| Phosphorus Assay Reagent | 40 | 0 | 0 |
| Phosphorus Assay Reagent | 200 | 200 | 200 |
| Standard | 0 | 40 | 0 |
3.3 Measurement
Mix well after addition.
Incubate in a 40°C water bath for 10 minutes.
Cool at room temperature for 10 minutes.
Transfer 200 μL of the mixture to a 96-well plate or micro glass cuvette.
Measure the absorbance at 660 nm.
Precautions
The Phosphorus Assay Reagent must be prepared fresh before use and used on the same day.
Complete the measurement within 40 minutes after starting the incubation step (3.3).
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.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ25F1230304 |
分析证书 | S1506787 |
危险品化学品经营许可证（带存储）