计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
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。
质量文档
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。
文献证明
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。
概述
过氧化氢（H₂O₂）是生物体内最常见的活性氧分子，是一种活性氧代谢的副产物，主要由SOD和XOD 等催化产生，由 CAT 和 POD 等催化降解。过氧化氢不仅是重要的活性氧之一，也是活性氧相互转化的枢纽。一方面，H₂O₂可以直接或间接地氧化细胞内核酸，蛋白质等生物大分子，使细胞膜遭受损害，从而加速细胞的衰老和解体；另一方面，H₂O₂也是许多氧化应急反应中的关键调节因子。H₂O₂可以激活NF-κB 等因子，这些过氧化氢相关的信号途径和哮喘、炎症性关节炎、动脉硬化以及神经退行性疾病等许多疾病相关。H₂O₂也和细胞凋亡、细胞增殖等密切相关。
检测原理：H₂O₂与硫酸钛反应生成的过氧化物-钛复合物黄色沉淀，溶解于强酸中，其黄色深浅与H₂O₂浓度在一定范围内呈线性关系，通过酶标仪微板法测定412nm处吸光度，主要用于检测植物组织、血清、血浆等样品中H₂O₂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。
| H1505503 | Component | 100T | Storage |
| H1505503A | H₂O₂基液 | 1 mL | 2-8℃. Store in the dark. |
| H1505503B | 碱性基液 | 10.5 mL | RT. |
| H1505503C | 硫酸钛 | 0.3 g | RT. |
| H1505503D | 酸性基液 | 100 mL | RT. |
自备仪器和试剂
1、蒸馏水、丙酮
2、匀浆器或研钵、低温离心机、酶标仪、96孔板
实验流程
1、样品准备
1.1 植物样品
取正常或逆境下的新鲜植物组织，清洗干净，擦干，切碎，迅速称取5g，加入5ml预冷的丙酮，在冰浴条件下迅速匀浆或研磨，4℃ 12000g离心20min，收集上清液，测量提取液总体积，4℃保存备用。
1.2 血浆、血清和尿液样品
血浆、血清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，用于过氧化氢的测定。
1.3 高活性样品
如果样品中含有较高浓度的过氧化氢，可以使用丙酮进行恰当的稀释。
2、10mM H₂O₂标准溶液的配制
由于H₂O₂不是非常稳定，使用前需自行测定H₂O₂的实际浓度，本试剂盒提供的H₂O₂基液的H₂O₂浓度约为1M，用蒸馏水稀释100倍，使H₂O₂浓度约为10mM，蒸馏水调零，分光光度计测定A<sub>240</sub>，根据公式H₂O₂浓度 (mM)=22.94×A<sub>240</sub>计算出H₂O₂基液中H₂O₂的实际浓度，再用丙酮稀释H₂O₂基液配制10mM H₂O₂-丙酮标准溶液（一般情况下新配制的10mM H₂O₂基液A<sub>240</sub>为0.4~0.45，3个月以后A<sub>240</sub>为0.35~0.42）。
按下表依次稀释（常用浓度0.3-3mM，即1~5号）：
| 序号 | H₂O₂-丙酮标准 (10mM) (ml) | 预冷丙酮 (ml) | H₂O₂浓度 (mM) |
| 1 | 0.03 | 0.97 | 0.3 |
| 2 | 0.05 | 0.95 | 0.5 |
| 3 | 0.08 | 0.92 | 0.8 |
| 4 | 0.1 | 0.9 | 1 |
| 5 | 0.3 | 0.7 | 3 |
| 6 | 0.5 | 0.5 | 5 |
| 7 | 0.8 | 0.2 | 8 |
3、硫酸钛溶液的配制
0.3g硫酸钛加入6ml蒸馏水中，即为5%硫酸钛溶液，4℃保存。
4、H₂O₂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H₂O₂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。
| 加入物 (ml) | 空白管 | 标准管 | 测定管 |
| 预冷丙酮 | 0.5 | — | — |
| 系列H₂O₂-丙酮溶液（1~5号） | — | 0.5 | — |
| 待测样品 | — | — | 0.5 |
| 碱性基液 | 0.1 | 0.1 | 0.1 |
| 硫酸钛溶液 | 0.05 | 0.05 | 0.05 |
注：加入碱性基液、硫酸钛溶液时，应直接加至溶液中，不要粘到管壁。
5、H₂O₂测定
混匀，室温放置5min，12000g离心15min，弃上清液，留取沉淀，如有必要可加入预冷丙酮反复洗涤沉淀物，向各管的沉淀中加入1ml酸性基液，摇动，使沉淀完全溶解，取各管溶液300μl加入96孔板，空白调零，酶标仪测定412nm处各标准孔、测定孔的吸光度。
6、结果计算
6.1 标准曲线的制作
以系列H₂O₂-丙酮标准（0.3、0.5、0.8、1、3、5、8 mM）为横坐标，以对应的吸光度为纵坐标，绘制标准曲线，求得回归方程；
6.2 样本H₂O₂浓度计算
以测定管吸光度代入回归方程求得待测样品中H₂O₂的浓度（C₀）。
组织样品H₂O₂ (mmol/g)=(C₀×VT×N)/m
液体样品H₂O₂ (mmol/L)=C₀×N
参数说明
C₀：根据待测样品的吸光度在标准曲线求得H₂O₂浓度 (mM)；
VT：待测样品的总体积 (L)；
m：样品质量 (g)；
N：样本稀释倍数。
结果展示
标准曲线制作：在室温条件下按说明书操作，对系列标准进行吸光度的测定，其吸光度及标准曲线如下（仅供参考），我们采用H₂O₂标准（0.1、0.3、0.5、0.8、1、3、5、8、10mM）绘制标准曲线（标准品浓度过高或过低都有可能影响标准曲线的准确性）：
| H₂O₂标准 (mM) | 0.1 | 0.3 | 0.5 | 0.8 | 1 | 3 | 5 | 8 | 10 |
| 吸光度 | 0.009 | 0.065 | 0.141 | 0.216 | 0.264 | 0.812 | 1.262 | 2.010 | 2.355 |
注意：H₂O₂浓度低于0.3mM基本无色，0.3~1mM为黄色，3~10mM为橙黄色，效果参考如下。
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注意事项
1、该试剂盒亦可用分光光度计进行检测，但检测的样本数相应减少。
2、加入碱性基液、硫酸钛溶液时，应直接加入至溶液中，不要粘到管壁。
3、过氧化物-钛复合物黄色沉淀溶解于酸性基液时需要一段时间，需完全溶解，否则有可能影响测定结果。
4、H₂O₂基液和碱性基液应严格密闭保存，避免挥发，否则效率会下降。
5、H₂O₂基液和酸性基液有一定腐蚀性，请小心操作。
6、硫酸钛溶解于水后应尽早使用，如暂时不用，可短期放置4℃冰箱保存；亦可用分析天平称取一定量的粉剂，配置5%的浓度即可。
7、为了您的安全和健康，请穿实验服并戴一次性手套操作。
8、试剂开封后请尽快使用，以防影响后续实验效果。
Hydrogen peroxide (H₂O₂) is the most common reactive oxygen species (ROS) molecule in living organisms. It is a byproduct of active oxygen metabolism, primarily generated by enzymes like SOD and XOD, and degraded by enzymes such as CAT and POD. H₂O₂ is not only a significant ROS but also a hub for the interconversion of various reactive oxygen species. On one hand, H₂O₂ can directly or indirectly oxidize biological macromolecules like nucleic acids and proteins within cells, damaging cell membranes and thereby accelerating cellular aging and disintegration. On the other hand, H₂O₂ is a key regulatory factor in many oxidative stress responses. It can activate factors like NF-κB, and these H₂O₂-related signaling pathways are associated with numerous diseases such as asthma, inflammatory arthritis, arteriosclerosis, and neurodegenerative disorders. H₂O₂ is also closely linked to processes like cell apoptosis and proliferation.
Assay Principle
H₂O₂ reacts with titanium sulfate to form a yellow peroxide-titanium complex precipitate. This precipitate dissolves in strong acid, and the intensity of the yellow color is linearly proportional to the H₂O₂ concentration within a certain range. The absorbance at 412 nm is measured using a microplate reader. This kit is primarily used to detect H₂O₂ content in samples such as plant tissues, serum, and plasma. It is intended for research use only and not suitable for clinical diagnosis or other purposes.
| H1505503 | Component | 100T | Storage |
| H1505503A | H₂O₂ Stock Solution | 1 mL | 2-8℃. Store in the dark. |
| H1505503B | Alkaline Matrix Solution | 10.5 mL | RT. |
| H1505503C | Titanium Sulfate | 0.3 g | RT. |
| H1505503D | Acidic Matrix Solution | 100 mL | RT. |
Required Materials Not Provided
1. Distilled water, Acetone
2. Homogenizer or mortar and pestle, Refrigerated centrifuge, Microplate reader, 96-well plate
Experimental Procedure
1. Sample Preparation
1.1 Plant Samples
Take fresh plant tissue (under normal or stress conditions), wash clean, dry, and mince.
Quickly weigh 5 g, add 5 mL of pre-chilled acetone.
Homogenize or grind rapidly in an ice bath.
Centrifuge at 12,000 g for 20 min at 4°C.
Collect the supernatant, measure the total volume of the extract, and store at 4°C for later use.
1.2 Plasma, Serum, and Urine Samples
Plasma and serum, prepared by conventional methods, can be directly used for assay with this kit. Store at 4°C for H₂O₂ determination.
1.3 High-Activity Samples
If the sample contains a high concentration of H₂O₂, it can be appropriately diluted with acetone.
2. Preparation of 10 mM H₂O₂ Standard Solution
Since H₂O₂ is not very stable, its actual concentration needs to be determined before use.
The H₂O₂ Stock Solution provided has an H₂O₂ concentration of approximately 1 M. Dilute it 100-fold with distilled water to achieve ~10 mM.
Zero the spectrophotometer with distilled water and measure the absorbance at 240 nm (A<sub>240</sub>).
Calculate the actual H₂O₂ concentration using the formula: H₂O₂ concentration (mM) = A<sub>240</sub> × 22.94.
Then, dilute the H₂O₂ Stock Solution with acetone to prepare a 10 mM H₂O₂-acetone standard solution.
Prepare serial dilutions according to the table below (common range 0.3-3 mM, i.e., Tubes 1-5):
| Tube | 10 mM H₂O₂-Acetone Standard (mL) | Pre-chilled Acetone (mL) | Final H₂O₂ Concentration (mM) |
| 1 | 0.03 | 0.97 | 0.3 |
| 2 | 0.05 | 0.95 | 0.5 |
| 3 | 0.08 | 0.92 | 0.8 |
| 4 | 0.1 | 0.9 | 1 |
| 5 | 0.3 | 0.7 | 3 |
| 6 | 0.5 | 0.5 | 5 |
| 7 | 0.8 | 0.2 | 8 |
3. Preparation of Titanium Sulfate Solution
Add 0.3 g of Titanium Sulfate to 6 mL of distilled water to make a 5% Titanium Sulfate solution. Store at 4°C.
4. H₂O₂ Assay Setup
Set up Blank, Standard, and Test tubes as per the table below.
Add reagents in the specified order, avoiding bubbles.
If the sample H₂O₂ concentration is too high, reduce the sample volume or dilute appropriately before assay. Duplicate tubes are recommended.
| Reagent (mL) | Blank Tube | Standard Tube | Test Tube |
| Pre-chilled Acetone | 0.5 | — | — |
| Serial H₂O₂-Acetone Standards | — | 0.5 | — |
| Sample Extract | — | — | 0.5 |
| Alkaline Matrix Solution | 0.1 | 0.1 | 0.1 |
| Titanium Sulfate Solution | 0.05 | 0.05 | 0.05 |
Note: When adding Alkaline Matrix Solution and Titanium Sulfate Solution, add them directly into the liquid, avoiding the tube walls.
5. H₂O₂ Measurement
Mix well and let stand at room temperature for 5 minutes.
Centrifuge at 12,000 g for 15 minutes.
Discard the supernatant carefully, retaining the precipitate.
If necessary, wash the precipitate repeatedly with pre-chilled acetone.
Add 1 mL of Acidic Matrix Solution to the precipitate in each tube.
Shake to dissolve the precipitate completely.
Transfer 300 µL of each tube's solution to a 96-well plate.
Zero the microplate reader using the blank solution.
Measure the absorbance of standard and test wells at 412 nm.
6. Calculation of Results
6.1 Standard Curve Plotting
Plot the standard curve using the serial H₂O₂-acetone standard concentrations (0.3, 0.5, 0.8, 1, 3, 5, 8 mM) on the X-axis and their corresponding absorbances on the Y-axis. Determine the regression equation.
6.2 Sample H₂O₂ Concentration Calculation
Substitute the absorbance of the test sample into the regression equation to find its H₂O₂ concentration (C₀).
For tissue samples: H₂O₂ (mmol/g weight) = (C₀ × V<sub>T</sub> × N) / m
For liquid samples: H₂O₂ (mmol/L) = C₀ × N
Parameter Definitions:
C₀: H₂O₂ concentration (mM) obtained from the standard curve based on the sample's absorbance.
VT: Total volume of the test sample extract (L).
m: Sample mass (g).
N: Sample dilution factor.
Results Presentation
Standard Curve Example: When measured according to the instructions at room temperature, the absorbance values for the serial standards and a typical standard curve are as follows (for reference only). A standard curve was plotted using H₂O₂ standards (0.1, 0.3, 0.5, 0.8, 1, 3, 5, 8, 10 mM). Note: Standard concentrations that are too high or too low may affect the accuracy of the standard curve.
| H₂O₂ Standard (mM) | 0.1 | 0.3 | 0.5 | 0.8 | 1 | 3 | 5 | 8 | 10 |
| Absorbance | 0.009 | 0.065 | 0.141 | 0.216 | 0.264 | 0.812 | 1.262 | 2.010 | 2.355 |
Note: H₂O₂ concentrations below 0.3 mM are basically colorless, 0.3~1 mM appear yellow, and 3~10 mM appear orange-yellow. Refer to the image below for expected results.
Precautions
1. This kit can also be used with a spectrophotometer, but the number of samples processed will be correspondingly reduced.
2. When adding Alkaline Matrix Solution and Titanium Sulfate Solution, add them directly into the solution, avoiding the tube walls.
3. Dissolving the yellow peroxide-titanium complex precipitate in the Acidic Matrix Solution takes time. Ensure it is completely dissolved, otherwise results may be affected.
4. The H₂O₂ Stock Solution and Alkaline Matrix Solution should be tightly sealed to prevent evaporation, as efficiency may decrease otherwise.
5. The H₂O₂ Stock Solution and Acidic Matrix Solution are corrosive. Handle with care.
6. The Titanium Sulfate solution should be used soon after preparation. If not used immediately, it can be stored short-term at 4°C. Alternatively, weigh an appropriate amount of powder and prepare a 5% solution as needed.
7. For your safety and health, wear lab coats and disposable gloves during operation.
8. Use reagents promptly after opening to avoid affecting subsequent experimental results.
规格
别名
过氧化氢检测试剂盒 | H₂O₂检测试剂盒，比色法
稳定性与储存
各组分在相应的储存条件下保质期1年。
英文名称
Hydrogen Peroxide (H₂O₂) Content Assay Kit (TTS, Micro Method)
储存条件
2-8°C储存,避光,室温
运输条件
低温运输
产品线
生物产品
名称和识别符
分子类型
试剂盒
技术文档
高级数据
化学和物理性质
敏感性
Light-sensitive
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C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
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 |
|---|---|---|
 H2621219 |
分析证书 | H1505503 |
| H2617025 |
分析证书 | H1505503 |
| ZJ26F0737698 |
分析证书 | H1505503 |
| F2611342 |
分析证书 | H1505503 |
| E2627308 |
分析证书 | H1505503 |
| ZJ26F0535519 |
分析证书 | H1505503 |
| D2623111 |
分析证书 | H1505503 |
| D2623119 |
分析证书 | H1505503 |
| D2613120 |
分析证书 | H1505503 |
| D2603173 |
分析证书 | H1505503 |
| C2611320 |
分析证书 | H1505503 |
| ZJ26F0232232 |
分析证书 | H1505503 |
| L2512320 |
分析证书 | H1505503 |
| L2503088 |
分析证书 | H1505503 |
| ZJ25F1129729 |
分析证书 | H1505503 |
技术文档和文章
溶液计算器
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