产品简介：
土壤中的酚类物质主要由植物释放，以及植物残体和凋落物分解产生。由于酚类物质的难降解性，其在土壤中的积累会影响土壤的碳氮转化和温室气体排放，进而会使土壤肥力衰退。另外，酚类物质对土壤有机质矿化和养分循环也有着重要影响。本试剂盒采用福林酚法测定土壤中酚类物质含量，在碱性条件下，酚类物质将钨钼酸还原，产生蓝色化合物，在750 nm处有特征吸收峰，通过检测在750 nm处的吸光值，进而计算土壤中酚类物质含量。
组份表：
| S1521998 | 组件 | 物理外观 | 48T | 96T | 储存 | 单次测试使用量 | | S1521998A | 提取液 | 无色透明液体 | 60 mL | 100 mL | 2-8℃. 避光 | 每次反应需使用1 mL | | S1521998B | 试剂一 | 黄色液体 | 7 mL | 14 mL | 2-8℃. 避光 | 每次反应需使用125 μL | | S1521998C | 试剂二 | 无色透明液体 | 1.5 mL | 2.5 mL | 2-8℃ | 每次反应需使用25 μL | | S1521998D | 标准品 | 白色粉末 | 10 mg | 10 mg | 2-8℃ | 按需求使用 |
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所需的仪器和用品：
酶标仪、96孔板、可调式移液器、天平、离心机。
土壤总酚含量的测定：
建议正式实验前选取2个样本做预测定，了解本批样品情况，熟悉实验流程，避免实验样本和试剂浪费！
1.样本制备：
称取约0.5 g土壤样本，加入1 mL提取液，室温振荡提取30 min。25℃×8000 rpm，离 心10 min，取上清待测。
2.上机检测：
(1)可见分光光度计预热30 min，调节波长至750 nm，蒸馏水调零。
(2)在96孔板中依次加入：
| 试剂名称(μL) | 测定管 | 空白管(仅做一次) | | 样本 | 50 | - | | 蒸馏水 | - | 50 | | 试剂一 | 125 | 125 | | 试剂二 | 25 | 25 |
混匀，25℃室温静置30 min,全部液体转移至96孔板中，测定750 nm吸光值A, △A=A测定-A A空白 |
结果计算：
1.标准曲线：Y=0.2325*X+0.04544，X是标准品质量(μg)，Y是∆A。
2.土壤总酚含量(μg/g土壤)=(∆A-0.04544)÷0.2325÷(V1÷V×W)×D
V---加入提取液体积，1 mL;
D---稀释倍数，未稀释即为1;
V1---反应中样品体积，0.05 mL;
W---土壤取样质量，g。
附 标准曲线制作过程：
1.制备标准品母液(10 mg/mL)：向标准品EP管里面加入1 mL蒸馏水，超声完全溶解。
2.把母液用蒸馏水稀释成五个浓度梯度的标准品：0，0.02，0.04，0.06，0.08，0.1 mg/mL。也可根据实际样本来调整标准品浓度。
操作表(吸取标准品母液50 μL，加入4.95 mL蒸馏水，混匀得到0.1 mg/mL的标品稀释液待用)：
| ① | ② | ③ | ④ | ⑤ | ⑥ | | 标品浓度(mg/mL) | 0 | 0.02 | 0.04 | 0.06 | 0.08 | 0.1 | | 标品稀释液(μL) | 0 | 80 | 160 | 240 | 320 | 400 | | 蒸馏水(μL) | 400 | 320 | 240 | 160 | 80 | 0 |
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3. 依据测定管的加样体系操作，根据结果即可制作标准曲线。
Background Introduction:
The phenolic substances in the soil are mainly released by plants and produced by the decomposition of plant residues and fallen leaves. Due to the difficulty in their degradation, the accumulation of phenolic substances in the soil can affect the carbon-nitrogen transformation and greenhouse gas emissions of the soil, thereby causing a decline in soil fertility. Back. In addition, phenolic substances also have significant effects on the mineralization of soil organic matter and the nutrient cycle. This kit uses the Folin phenol method to determine the content of phenolic substances in the soil. Under alkaline conditions, phenolic substances will reduce tungsten molybdate, generating a blue compound. There is a characteristic absorption peak at 750 nm. By detecting the absorbance value at 750 nm, the content of phenolic substances in the soil can be calculated.
Components:
| S1521998 | Components | Appearance | 48T | 96T | Storage | Quantity Per Test | | S1521998A | Extraction Solution | Colorless clear liquid | 60 mL | 100 mL | 2-8℃. Store in the dark | 1 mL for 1 reaction | | S1521998B | Reagent 1 | Yellow liquid | 7 mL | 14 mL | 2-8℃. Store in the dark | 125 μL for 1 reaction | | S1521998C | Reagent 2 | Colorless clear liquid | 1.5 mL | 2.5 mL | 2-8℃ | 25 μL for 1 reaction | | S1521998D | Standard Sample | White powder | 10 mg | 10 mg | 2-8℃ | Use as per requirements |
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Required instruments and supplies:
Enzyme analyzer, 96-well plate, adjustable pipette, balance, centrifuge.
Determination of total phenolic content in soil:
It is recommended to select 2 samples for prediction before the formal experiment. This will help us understand the situation of this batch of samples, familiarize ourselves with the experimental process, and avoid waste of experimental samples and reagents!
1. Sample preparation:
Take approximately 0.5 g of soil sample, add 1 mL of extraction solution, and shake at room temperature for 30 minutes. Centrifuge at 25℃ and 8000 rpm for 10 minutes, then take the supernatant for testing.
2. Machine testing:
(1) The spectrophotometer was preheated for 30 minutes, the wavelength was adjusted to 750 nm, and distilled water was used for zero adjustment.
(2) Add successively in the 96-well plate:
| Reagent Name (μL) | Testing tube | Blank tube (used only once) | | Sample | 50 | - | | Distilled water | - | 50 | | Reagent 1 | 125 | 125 | | Reagent 2 | 25 | 25 |
Mix well, let it stand at room temperature (25℃) for 30 minutes. Transfer all the liquid to the 96-well plate. Measure the absorbance at 750 nm as A, and calculate △A = A(measured) - A(blank) |
Result Calculation:
1. Standard curve: Y = 0.2325*X + 0.04542, where X represents the mass of the standard sample (in μg) and Y represents ΔA.
2. Total phenol content in soil (μg/g soil) = (∆A - 0.04542) ÷ 0.2325 ÷ (V1 ÷ V × W) × D
V---Volume of the extracted liquid, 1 mL;
D---Dilution factor, 1 for undiluted sample;
V1---Volume of the sample in the reaction, 0.05 mL;
W---Mass of soil sample, g.
Appendix Procedure for preparing the standard curve:
1. Prepare standard stock solution (10 mg/mL): Add 1 mL distilled water to the standard powder vial, sonicate until fully dissolved.
2. Dilute the mother solution with distilled water to prepare five concentration gradients of standard samples: 0, 0.02, 0.04, 0.06, 0.08, 0.1 mg/mL. The concentration of the standard substance can also be adjusted according to the actual sample.
Operation Table (Take 50 μL of the standard solution stock solution, add 4.95 mL of distilled water, and mix well to obtain a 0.1 mg/mL standard dilution solution for later use)：
| ① | ② | ③ | ④ | ⑤ | ⑥ | | Standard concentration (mg/mL) | 0 | 0.02 | 0.04 | 0.06 | 0.08 | 0.1 | | Standard diluent (μL) | 0 | 80 | 160 | 240 | 320 | 400 | | Distilled water (μL) | 400 | 320 | 240 | 160 | 80 | 0 |
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3. Operate following the pipetting protocol for test tubes above, then plot the standard curve using recorded absorbance values.
ZJ26F0838361
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