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。
果糖是一种最为常见的己酮糖，是葡萄糖的同分异构体，以游离状态大量存在于水果的浆汁和蜂蜜中，能与葡萄糖结合生成蔗糖。果糖是最甜的单糖，广泛应用于食品、医药、保健品生产中。
检测原理：在酸性条件下果糖与间苯二酚反应，生成有色物质，在480nm下有特征吸收峰。
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
操作步骤
1、试剂准备
| 试剂名称 | 试剂准备 | 备注 |
| Extraction Buffer | 即用型，使用前平衡到室温 | 4℃保存 |
| Working Reagent Ⅰ | 临用前配制；加入1mL去离子水，充分溶解，取100μL上述溶液加入900μL去离子水充分溶解备用。 | 用不完的试剂4℃避光可保存两周 |
| Reagent Ⅱ | 即用型，使用前平衡到室温 | 4℃保存 |
| Reagent Ⅲ | 即用型，使用前平衡到室温 | 4℃避光保存 |
| Reagent Ⅳ | 即用型 | 常温避光保存 |
注：Reagent Ⅱ和Reagent Ⅲ有毒且有刺激性气味，建议在通风橱进行实验。
2、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4h内完成样品解冻。
称取0.1-0.2 g 样本，常温研碎；加入0.5mL Extraction Buffer，适当研磨后快速转移到有盖离心管中；置于80℃水浴锅中10min （盖紧，以防止水分散失），振荡3-5次，冷却后，4,000g，25℃离心10min，取上清；加入少量（约2mg）Reagent Ⅳ，80℃脱色30min（盖紧，以防止水分散失）；再加入0.5mL Extraction Buffer，4,000g，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480nm。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 EP管中进行）：
| 试剂（μL） | 测定管 | 空白管 | 标准管 |
| 样本上清 | 30 | 0 | 0 |
| Working Reagent Ⅰ | 0 | 0 | 30 |
| 去离子水 | 0 | 30 | 0 |
| Reagent Ⅱ | 210 | 210 | 210 |
| Reagent Ⅲ | 60 | 60 | 60 |
3.3 混匀，95℃水浴反应30min（盖紧，以防止水分散失），冷却后取200μL至微量玻璃比色皿或96孔板中测定480nm处吸光值，分别记为A测定、A空白、A标准，计算ΔA测定=A测定-A空白、ΔA标准=A标准-A空白。
注意：空白管和标准管只需做1-2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A测定小于0.05可适当加大样本量。如果A测定大于2.0，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
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。建议以加粗的简洁计算公式为最终计算公式。
果糖含量 (mg/g 鲜重)=(C标准×V样)×ΔA测定÷ΔA标准÷(W×V样÷V样总)=ΔA测定÷ΔA标准÷W
参数说明：
C标准：标准管浓度，1mg/mL；
V样：加入样本体积，0.03mL；
V样总：加入提取液体积，1mL；
W：样本鲜重，g。
5、结果展示
图1. 本试剂盒测定香瓜和橘子中果糖的含量
注意事项
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。
Fructose is one of the most common ketohexoses and an isomer of glucose. It exists in large amounts in the free state in fruit juices and honey and can combine with glucose to form sucrose. Fructose is the sweetest monosaccharide and is widely used in the production of food, pharmaceuticals, and health products.
Detection Principle
Under acidic conditions, fructose reacts with resorcinol to produce a colored substance with a characteristic absorption peak at 480 nm.
Note: Before formal testing, it is recommended to perform a preliminary experiment with 2-3 samples expected to show significant differences.
Reagents, consumables and Equipments not provided
Microplate reader or visible spectrophotometer (capable of measuring absorbance at 480 nm)
96-well plate or micro-volume glass cuvette, Adjustable pipettes and tips
Constant temperature water bath, Centrifuge, Analytical balance
Deionized water
Homogenizer
Procedure
1. Reagent Preparation
| Reagent Name | Preparation | Notes |
| Extraction Buffer | Ready-to-use. Equilibrate to room temperature before use. | Store at 4°C. |
| Working Reagent Ⅰ | Prepare immediately before use. Dissolve Reagent Ⅰ in 1 mL deionized water. Take 100 µL of this solution and add to 900 µL deionized water. Mix thoroughly. | Unused reagent can be stored at 4°C protected from light for up to two weeks. |
| Reagent Ⅱ | Ready-to-use. Equilibrate to room temperature before use. | Store at 4°C. |
| Reagent Ⅲ | Ready-to-use. Equilibrate to room temperature before use. | Store at 4°C protected from light. |
| Reagent Ⅳ | Ready-to-use. | Store at room temperature protected from light. |
Note: Reagent Ⅱ and Reagent Ⅲ are toxic and have an irritating odor. It is recommended to perform the experiment in a fume hood.
2. Sample Preparation
*Note: Fresh samples are recommended. If not used immediately, samples can be stored at -80°C for several weeks. Control the thawing temperature and time during measurement. When thawing at room temperature, complete the process within 4 hours.*
Weigh 0.1-0.2 g of sample and grind at room temperature. Add 0.5 mL of Extraction Buffer, grind appropriately, and quickly transfer to a capped centrifuge tube. Place in an 80°C water bath for 10 minutes (keep cap tight to prevent water loss), vortex 3-5 times. After cooling, centrifuge at 4,000 g, 25°C for 10 minutes. Collect the supernatant. Add a small amount (approx. 2 mg) of Reagent Ⅳ. Decolorize at 80°C for 30 minutes (keep cap tight). Add another 0.5 mL of Extraction Buffer, centrifuge at 4,000 g, 25°C for 10 minutes. Collect the supernatant for testing.
3. Experimental Steps
3.1 Preheat the microplate reader or visible spectrophotometer for 30 minutes and set the wavelength to 480 nm. Zero the visible spectrophotometer with deionized water.
3.2 Operation Table (perform in 1.5 mL EP tubes):
| Reagent (µL) | Test Tube | Blank Tube | Standard Tube |
| Sample Supernatant | 30 | 0 | 0 |
| Working Reagent Ⅰ | 0 | 0 | 30 |
| Deionized Water | 0 | 30 | 0 |
| Reagent Ⅱ | 210 | 210 | 210 |
| Reagent Ⅲ | 60 | 60 | 60 |
3.3 Mix well. React in a 95°C water bath for 30 minutes (keep cap tight to prevent water loss). After cooling, transfer 200 µL to a micro-volume glass cuvette or 96-well plate. Measure the absorbance at 480 nm, recorded as ATest, ABlank, AStandard. Calculate ΔATest = ATest - ABlank and ΔAStandard = AStandard - ABlank.
*Note: The blank and standard tubes only need to be prepared 1-2 times. It is recommended to perform a preliminary experiment with 2-3 samples showing expected large differences before formal testing. If A<sub>Test</sub> is less than 0.05, appropriately increase the sample amount. If A<sub>Test</sub> is greater than 2.0, further dilute the sample with Extraction Buffer and multiply the result by the dilution factor, or reduce the amount of sample used for extraction.*
4. Calculation of Results
Note: We provide two formula sets: the derived calculation formulas and the simplified formulas. They are completely equivalent. The simplified formulas are recommended as the final calculation formulas.
Fructose Content Calculation:
Fructose Content (mg/g fresh weight) = (CStandard × VSample) × ΔATest ÷ ΔAStandard ÷ (W × VSample ÷ VTotal Extract) = ΔATest ÷ ΔAStandard ÷ W
Parameter Description:
CStandard: Standard tube concentration, 1 mg/mL;
VSample: Volume of sample added, 0.03 mL;
V<sub>Total Extract</sub>: Volume of extraction buffer added, 1 mL;
W: Sample fresh weight, g.
5. Example Results
Figure 1. Fructose content in melon and orange determined by this kit
Notes
This product is for scientific research use only and is not intended for clinical diagnosis. For your safety and health, please wear a lab coat and disposable gloves during operation.
| P1508220 | Component | 48T | 96T | Storage |
| P1508220A | Extraction Buffer | 60 mL | 120 mL | 2-8℃ |
| P1508220B | Reagent Ⅰ | 1EA | 1EA | 2-8℃. Store in the dark. |
| P1508220C | Reagent Ⅱ | 13 mL | 26 mL | 2-8℃ |
| P1508220D | Reagent Ⅲ | 4 mL | 8 mL | 2-8℃. Store in the dark. |
| P1508220E | Reagent Ⅳ | 1EA | 1EA | RT. Store in the dark. |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 J2628571 |
分析证书 | P1508220 |
| ZJ26F0131079 |
分析证书 | P1508220 |
| ZJ26F0131080 |
分析证书 | P1508220 |
| ZJ26F0131081 |
分析证书 | P1508220 |
| ZJ26F0131082 |
分析证书 | P1508220 |
| ZJ26F0131083 |
分析证书 | P1508220 |
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