BioReagent 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
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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。
植物组织或果实中存在花青素、叶绿素、类胡萝卜素、类黄酮、酚类等物质，这些物质与果实等样品衰老过程密切相关，对其加工性能、存储、营养价值等都有重要影响，植物酚类物质具有清除自由基，抗氧化抗衰老的作用，具有较高的营养价值和医疗保健作用而广泛应用于化妆品、食品、医药等领域。
检测原理：总酚（Total Phenol）溶于有机溶剂，以有机溶剂粗提总酚，根据提取液的吸收光谱特性，可利用紫外分光光度计在280nm处测定其吸光度，通过与标准曲线比较，计算出总酚含量，主要用于植物组织或果实中总酚的提取以及定量检测总酚含量。该试剂盒仅用于科研领域，不宜用于临床诊断或其他用途。
适用样本：植物组织、果实
自备仪器和试剂
天平、称量纸、剪刀、研钵或匀浆器、离心管、离心机、滤纸或纱布、石英比色杯
操作步骤（仅供参考）
1、总酚提取
1.1 取果实或其他植物组织，洗净，擦干，称取剪碎的新鲜样品0.25g，置于4℃预冷的研钵或匀浆器。
1.2 加入4℃预冷的2~3mL TP Assay Buffer，充分研磨或匀浆后转入10mL离心管，用TP Assay Buffer冲洗研钵或匀浆器并转移至离心管，补加TP Assay Buffer至8mL。
1.3 4℃避光静置20min，期间摇动2~3次，然后过滤至离心管，也可用离心机8000r/min离心3min，滤液（上清液）即为总酚粗提液。
2、稀释总酚标准溶液
将总酚标准 (1mg/mL) 用TP Assay Buffer稀释成总酚标准 (50μg/mL)，取干净离心管或试管，按下表进行操作，依次稀释。
加入物 | 1 | 2 | 3 | 4 | 5 | 6 |
总酚标准 (50μg/mL) (mL) | 0.1 | 0.2 | 0.3 | 0.4 | 0.5 | 0.6 |
TP Assay Buffer (mL) | 0.9 | 0.8 | 0.7 | 0.6 | 0.5 | 0.4 |
总酚浓度 (μg/mL) | 5 | 10 | 15 | 20 | 25 | 30 |
3、TP加样
按照下表设置空白管、对照管、测定管；如果样品中的总酚浓度过高，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2平行管，求平均值。
加入物（mL） | 空白管 | 标准管 | 测定管 |
TP Assay Buffer | 1 | — | — |
系列总酚标准（1~6号管） | — | 1 | — |
总酚粗提液 | — | — | 1 |
注意：如用较大的比色杯，可相应增加试剂量，其后计算公式中V1=加样时所用总酚粗提液的体积（mL）应相应发生变化。
4、TP测定
各管试剂加入光径1cm的石英比色杯中，在5~15min内，以空白管调零，用紫外分光光度计280nm处测定系列标准管、测定管的吸光度。
5、结果计算
以1~6号管系列总酚标准浓度（5、10、15、20、25、30μg/mL）为横坐标，以对应的吸光度为纵坐标，绘制标准曲线，进而计算出各测定管总酚浓度（μg/mL）。
组织样品的总酚 (μg/g)=C×VT×N/W
液体样品的总酚 (μg/mL)=C×N
参数说明：
C：根据标准曲线求得的测定管总酚浓度 (μg/mL)
VT：总酚粗提液总体积 (ml) =8
W：样品鲜重 (g)
注意事项
1、为了避免总酚见光分解，操作时应尽量避光，研磨或匀浆时应尽量缩短时间。
2、取样量、试剂用量应根据总酚含量适当调整。
3、TP Assay Buffer应密闭保存，避免有效成分挥发。
4、测定时，石英比色皿上应加盖，防止挥发。
5、测定应及时，时间延长，总酚浓度高时，测定结果可能偏低。
6、植物总酚浓度在10~20ug/mL范围时，OD值基本稳定，超出范围时结果随时间有稍微变化。
7、如果没有紫外分光光度计，也可以使用全波长酶标仪测定。
8、为了您的安全和健康，请穿实验服并戴一次性手套操作。
9、试剂开封后请尽快使用，以防影响后续实验效果。
Plant tissues or fruits contain substances such as anthocyanins, chlorophyll, carotenoids, flavonoids, and phenolics. These substances are closely related to the aging process of fruits and other samples, significantly impacting their processing properties, storage, and nutritional value. Plant phenolics possess free radical‑scavenging, antioxidant, and anti‑aging effects. Due to their high nutritional value and health‑promoting properties, they are widely used in cosmetics, food, medicine, and other fields.
Detection Principle
Total phenols are soluble in organic solvents. Crude extraction of total phenols is performed using an organic solvent. Based on the absorption spectral characteristics of the extract, the absorbance can be measured at 280 nm using a UV spectrophotometer. By comparison with a standard curve, the total phenol content is calculated. This kit is primarily used for the extraction of total phenols from plant tissues or fruits and the quantitative determination of total phenol content. This kit is for research use only and is not suitable for clinical diagnosis or other purposes.
Applicable Samples: Plant tissues, fruits
Reagents, consumables and Equipments not provided
Operating Steps (for reference only)
1. Total Phenol Extraction
1.1 Take fruit or other plant tissue, wash and dry it. Weigh 0.25 g of fresh, cut‑up sample and place it in a mortar or homogenizer pre‑cooled at 4°C.
1.2 Add 2–3 mL of TP Assay Buffer (pre‑cooled at 4°C). Grind or homogenize thoroughly, then transfer to a 10 mL centrifuge tube. Rinse the mortar/homogenizer with TP Assay Buffer and transfer the rinse to the tube. Adjust the final volume to 8 mL with TP Assay Buffer.
1.3 Incubate at 4°C in the dark for 20 minutes, shaking 2–3 times during incubation. Then filter into a centrifuge tube. Alternatively, centrifuge at 8000 rpm for 3 minutes. The filtrate (supernatant) is the crude total phenol extract.
2. Dilution of Total Phenol Standard Solution
Dilute the Total Phenol Standard (1 mg/mL) with TP Assay Buffer to obtain a Total Phenol Standard (50 μg/mL). Using clean centrifuge tubes or test tubes, perform serial dilutions according to the following table:
Tube No. | 1 | 2 | 3 | 4 | 5 | 6 |
Total Phenol Standard (50 μg/mL) (mL) | 0.1 | 0.2 | 0.3 | 0.4 | 0.5 | 0.6 |
TP Assay Buffer (mL) | 0.9 | 0.8 | 0.7 | 0.6 | 0.5 | 0.4 |
Total Phenol Concentration (μg/mL) | 5 | 10 | 15 | 20 | 25 | 30 |
3. TP Sample Addition
Set up blank, standard, and test tubes according to the table below. If the total phenol concentration in the sample is too high, reduce the sample volume or dilute appropriately before measurement. It is recommended to set up duplicate tubes for each sample and take the average.
Substance Added (mL) | Blank Tube | Standard Tube | Test Tube |
TP Assay Buffer | 1 | — | — |
Total Phenol Standard Series (Tubes 1–6) | — | 1 | — |
Crude Total Phenol Extract | — | — | 1 |
Note: If using a larger cuvette, proportionally increase all reagent volumes. In this case, the volume V₁ of crude extract used in the calculation formula should be adjusted accordingly.
4. TP Measurement
Transfer the solutions from each tube to 1‑cm path length quartz cuvettes. Within 5–15 minutes, zero the spectrophotometer with the blank tube and measure the absorbance of the standard series and test tubes at 280 nm.
5. Result Calculation
Using the total phenol standard series concentrations (5, 10, 15, 20, 25, 30 μg/mL) as the x‑axis and the corresponding absorbance values as the y‑axis, plot a standard curve. Then calculate the total phenol concentration (μg/mL) for each test tube.
For tissue samples:
Total Phenol (μg/g) = C × VT × N / W
For liquid samples:
Total Phenol (μg/mL) = C × N
Parameter Explanation:
C: Total phenol concentration of the test tube obtained from the standard curve (μg/mL)
VTT: Total volume of crude total phenol extract (mL) = 8
W: Fresh weight of the sample (g)
N: Dilution factor
Precautions
To avoid photodegradation of total phenols, perform operations in low light as much as possible, and keep grinding/homogenization time short.
Sample amount and reagent volumes should be adjusted appropriately based on the total phenol content.
TP Assay Buffer should be stored tightly sealed to prevent evaporation of active components.
During measurement, cover the quartz cuvette to prevent evaporation.
Measurements should be performed promptly. Prolonged delay, especially with high phenol concentrations, may result in underestimation.
When plant total phenol concentration is within 10–20 μg/mL, the OD value is relatively stable. Outside this range, results may change slightly over time.
If a UV spectrophotometer is unavailable, a full‑wavelength microplate reader can also be used.
For your safety and health, please wear a lab coat and disposable gloves during operation.
Please use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0333561 |
分析证书 | P1509321 |
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