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。低温运输 。请查阅批次 COA 获取详细规格。
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
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。
在酸性溶液中，在分解剂和高温条件下，无机磷酸盐和有机磷酸盐会水解成正磷酸盐，正磷酸盐可与钼酸铵反应生成磷钼酸，在还原剂存在时被还原为磷钼蓝，其在 710 nm 处有特征吸收峰。
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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。
自备耗材
·酶标仪或可见分光光度计（能测 710 nm 处的吸光度）
·96 孔板或微量玻璃比色皿、可调节式移液枪及枪头
·台式离心机、30-50 目筛、恒温水浴锅
·去离子水
试剂准备
试剂一：即用型；使用前，平衡到室温； 2-8 ℃保存。
试剂二：即用型；使用前，平衡到室温； 2-8 ℃保存。
试剂三：即用型；使用前，平衡到室温； 2-8 ℃避光保存。
试剂四：临用前配制，加入 5.6 mL 去离子水充分溶解，然后加入试剂五充分混匀；试剂四含有少量不溶物，使用前需摇匀， 2-8 ℃保存 1 周。
试剂五：即用型；使用前，平衡到室温； 2-8 ℃保存。
注意：试剂一、试剂二、试剂三和试剂五有一定的刺激性，建议在使用过程中做好个人防护。
标准品：即用型；使用前，平衡到室温； 2-8 ℃保存。
标准品制备：使用 1 μmol/mL 标准品，按照下表所示，进一步稀释标准品：
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注意：每次实验都要做一次标准品检测，制作标准曲线；稀释后的标准品溶液不稳定，必须在 4 小时内使用。
样本制备
注意：推荐使用新鲜样本，如果不立即进行实验，样本可在-80℃保存数周。测定时，应控制解冻的温度和时间。室温环境下解冻
时，需在 4 h 内完成样品解冻。
1. 新鲜土样风干后过30-50目筛，按照土样质量（g）：去离子水体积(mL)：试剂一体积（mL）：试剂二体积（mL）=1:10:1:2
（建议称取约0.1 g土样，加入1 mL去离子水，再加入100 μL 试剂一和200 μL试剂二），95℃沸水浴振荡30 min，10,000g，
25℃离心10 min，取上清液待测。
实验步骤
1. 酶标仪或可见分光光度计预热 30 min，调节波长到 710 nm。可见分光光度计用去离子水调零。
2. 操作表（下述操作在 96 孔板或微量玻璃比色皿中进行）：
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3. 混匀后 25℃静置 10 min，于 710 nm 处测定吸光度，分别记为A空白、 A标准 、 A测定，计算 ΔA测定 = A测定 - A空白 、ΔA标准 = A标准 - A空白 。
结果计算
注意：我们为您提供的计算公式，包括推导过程计算公式和简洁计算公式。两者完全相等。
1. 标准曲线的绘制：
免责声明
In acidic solution, under the action of decomposing agent and high temperature, inorganic phosphate and organic phosphate are hydrolyzed into orthophosphate. Orthophosphate reacts with ammonium molybdate to form phosphomolybdic acid, which is then reduced to phosphomolybdenum blue in the presence of a reducing agent, with a characteristic absorbance peak at 710 nm.
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Notes: Before formal detection, it is recommended to conduct preliminary trials with 2–3 samples with expected large differences in phosphate content.
Self-Prepared Consumables and Instruments
1.Microplate reader or visible spectrophotometer (capable of absorbance detection at 710 nm)
2.96-well microplate or micro glass cuvettes, adjustable pipettes and pipette tips
3.Tabletop centrifuge, 30–50 mesh sieve, thermostatted water bath
4.Deionized water
Reagent Preparation
Reagent I: Ready-to-use; equilibrate to room temperature before use; store at 2-8 °C.
Reagent II: Ready-to-use; equilibrate to room temperature before use; store at 2-8 °C.
Reagent III: Ready-to-use; equilibrate to room temperature before use; store at 2-8 °C protected from light.
Working Reagent IV: Prepare immediately before use. Add 5.6 mL deionized water and dissolve completely, then add Reagent V and mix thoroughly. Working Reagent IV contains a small amount of insoluble precipitate; shake well before use. Stable for 1 week when stored at 2-8 °C.
Reagent V: Ready-to-use; equilibrate to room temperature before use; store at 2-8 °C.
Notes: Reagent I, Reagent II, Reagent III and Reagent V are irritant. Adequate personal protective equipment must be worn during all operations.
Standard Solution: Ready-to-use; equilibrate to room temperature before use; store at 2-8 °C.
Standard Solution Preparation: Dilute the 1 μmol/mL standard stock solution stepwise as shown in the table below.
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Notes: A standard curve must be generated alongside each independent experiment. Diluted standard solutions are unstable and must be used within 4 hours of preparation.
Sample Preparation
Fresh samples are recommended for immediate detection. If testing cannot be performed promptly, samples can be stored at −80 °C for several weeks. Control thawing temperature and duration strictly; complete thawing within 4 hours at room temperature.
1.Air-dry fresh soil samples and sieve through a 30–50 mesh screen.
Mix soil mass (g) : deionized water volume (mL) : Reagent I volume (mL) : Reagent II volume (mL) = 1:10:1:2.
Practical operation: Weigh approximately 0.1 g soil sample, add 1 mL deionized water, followed by 100 μL Reagent I and 200 μL Reagent II. Shake in a 95 °C boiling water bath for 30 min, centrifuge at 10,000 g, 25 °C for 10 min, collect supernatant for subsequent detection.
Assay Procedure
1.Preheat the microplate reader or visible spectrophotometer for ≥30 min, set detection wavelength to 710 nm. Zero the spectrophotometer with deionized water if applicable.
2.Assay layout (operations performed in 96-well microplate or micro glass cuvettes):
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3. Mix thoroughly and incubate at 25 °C for 10 min. Measure absorbance at 710 nm and record the absorbance values as Ablank, Astandard and Asample respectively.
Result Calculation
Notes: Two equivalent formulas are provided, namely the full derivation formula and the simplified formula.
Figure 1. Total phosphate content in soil detected by this kit
Disclaimer
This product is for research purposes only and is not suitable for clinical diagnosis. Please wear a lab coat and disposable gloves during operation to ensure your safety and health.
| 项目号 | 物理外观 | 组件 | 48T | 96T | 储存 |
| S1522127A | 液体 | 试剂一 | 10 mL | 20 mL | 2-8 ℃ |
| S1522127B | 液体 | 试剂二 | 15 mL | 30 mL | 2-8 ℃ |
| S1522127C | 液体 | 试剂三 | 2.8 mL | 5.6 mL | 2-8 ℃.避光 |
| S1522127D | 粉末 | 试剂四 | 1瓶 | 1瓶 | 2-8 ℃ |
| S1522127E | 液体 | 试剂五 | 44.8 µL | 44.8 µL | 2-8 ℃ |
| S1522127F | 液体 | 标准品 | 1 mL | 1 mL | 2-8 ℃ |
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