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。
植物体内的可溶性糖主要是指能溶于水及乙醇的单糖和寡聚糖，植物体内的碳素营养状况以及农产品的品质、性状，常以糖含量作为重要指标，植物为了适应逆境条件如干旱、低温等条件会主动积累一些可溶性糖，降低渗透势和冰点，以适应外界环境条件的变化，测定植物体内可溶性糖的方法有：蒽酮比色法、3,5-二硝基水杨酸法、苯酚比色法、斐林试剂比色法等化学方法。
检测原理：还原糖在浓硫酸作用下，可经脱水反应生成糖醛或羟甲基糠醛，生成物可与蒽酮反应生成蓝绿色糠醛衍生物，在一定范围内颜色的深浅与还原糖的含量成正比，在630nm处有最大吸收峰，本法几乎可以测定样品中所有的碳水化合物，不但可以测定戊糖（木糖、核糖、阿拉伯糖）、己糖（葡萄糖、果糖、山梨糖、半乳糖）、蔗糖、糖原、多缩葡萄糖，还可以测定所有的寡糖类和多糖，包括淀粉、纤维素等，实际上本产品可以一次性测定样本中所有碳水化合物的总量，在没有细分各物质的情况下可省去很多麻烦，具有特殊的应用价值。
该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。
| P1507813 | Component | 50T | Storage |
| P1507813A | 蔗糖标准溶液 (10mg/mL) | 1 mL | 2-8℃ |
| P1507813B | 蒽酮试剂 | 30 mL | 2-8℃. Store in the dark. |
自备仪器和试剂
1、蒸馏水、浓硫酸
2、电子天平、水浴锅或电磁炉、分光光度计、比色杯、剪刀、研钵或匀浆器
3、50ml烧杯或三角瓶、容量瓶、20ml刻度试管或10ml螺旋盖离心管
操作步骤
1、可溶性糖的提取
1.1 称取新鲜的植物样品（干样品亦可）0.5~1g，剪碎，加入蒸馏水约3ml匀浆，转移至刻度试管中，用12ml蒸馏水冲洗研磨器2~3次，洗出液也转移至该容器。
1.2 塑料薄膜封口，于沸水浴中提取30min，待冷却后过滤，将滤液转入50ml容量瓶。
1.3 收集残渣再次匀浆、加水提取、合并滤液，定容。
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3、加样
取10ml螺旋盖离心管，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，小心混匀；如果样品中的糖浓度过高，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2~3平行管，求平均值（各种试剂的加入量可以等比例的缩小，但应保证最小的所需量）。
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充分振荡，沸水浴中煮沸1min，取出，自然冷却至室温。
4、可溶性糖测定
混匀，以空白管调零，比色杯光径1cm，分光光度计测定630nm处标准管、测定管的吸光度。
5、结果计算
以系列蔗糖标准(1~5号)的质量(μg)为横坐标，以相应的吸光度为纵坐标，绘制标准曲线并求出线性回归方程，根据测定管的吸光度计算出相应的可溶性糖的质量及含量；亦可根据蔗糖标准浓度(μg/ml)与吸光度值绘制标准曲线，并求出样品的可溶性糖浓度。可溶性糖的含量，以质量分数(%)表示：
可溶性糖含量(%)=(m1×VT×N)/(m0×VS×1000000)×100%
=(c×VT×N)/(m0×1000000)×100%
参数说明：
m1：从标准曲线查得的可溶性糖的质量(μg)
VT：提取液的总体积(ml)
N：样品提取液的稀释倍数
m0：植物样品的质量(g)
VS：测定时所取样品提取液体积(ml)
c：样品的可溶性糖浓度(μg/ml)
1000000：μg与g的换算关系
注意事项
1、测定液必须清澈透明，加热后不应有蛋白沉淀，样品颜色较深时可用活性炭脱色后再进行测定。
2、如果样品可溶性糖浓度过高，应用蒸馏水稀释，糖的浓度在10~100ug/ml为宜。
3、浓硫酸(相对密度1.84)有强氧化性、强腐蚀性，危险性极大，操作应十分小心；加浓硫酸时应缓慢加入，以免产生大量热量而爆沸，灼伤皮肤和衣服，如出现此类现象，应迅速用自来水冲洗，如有必要应及时就医。
4、此方法测定结果受硫酸浓度和加热时间影响，操作时应准确、认真。
5、不同糖类与蒽酮试剂显色深度不同，果糖最深，葡萄糖次之，半乳糖、甘露糖较浅，五碳糖更浅。故测定糖的混合物时，常因不同糖类的比例不同造成误差，对于单一糖类的测定则不存在此误差。
附录
标准曲线制作：按说明书操作，通过分光光度计对系列标准进行吸光度的测定，其数值及标准曲线如下（仅供参考）：
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由结果可知，蔗糖标准在10~100μg/ml以内线性梯度良好。
Soluble sugars in plants mainly refer to monosaccharides and oligosaccharides that are soluble in water and ethanol. The carbon nutritional status in plants and the quality and characteristics of agricultural products are often evaluated based on sugar content. To adapt to adverse conditions such as drought and low temperature, plants actively accumulate soluble sugars to lower osmotic potential and freezing point, thereby adjusting to environmental changes. Methods for determining soluble sugars in plants include chemical approaches such as the anthrone colorimetric method, 3,5‑dinitrosalicylic acid method, phenol‑sulfuric acid method, and Fehling's reagent colorimetric method.
Detection Principle:
Under the action of concentrated sulfuric acid, reducing sugars undergo dehydration to form furfural or hydroxymethylfurfural. These products react with anthrone to produce a blue‑green furfural derivative. Within a certain range, the intensity of the color is proportional to the content of reducing sugars, with a maximum absorption peak at 630 nm. This method can detect almost all carbohydrates in a sample. It is applicable not only to pentoses (xylose, ribose, arabinose), hexoses (glucose, fructose, sorbose, galactose), sucrose, glycogen, and polyglucose, but also to all oligosaccharides and polysaccharides, including starch and cellulose. In practice, this product can measure the total carbohydrate content in a sample in a single assay, eliminating the need for separate measurements of individual components and offering unique application value.
This kit is intended for research use only and is not suitable for clinical diagnosis or other purposes.
| P1507813 | Component | 50T | Storage |
| P1507813A | Sucrose Standard Solution (10 mg/mL) | 1 mL | 2-8℃ |
| P1507813B | Anthrone Reagent | 30 mL | 2-8℃. Store in the dark. |
Reagents, consumables and Equipments not provided
1. Distilled water, concentrated sulfuric acid
2. Electronic balance, water bath or hot plate, spectrophotometer, cuvette, scissors, mortar or homogenizer
3. 50 mL beaker or conical flask, volumetric flask, 20 mL graduated test tubes or 10 mL screw-cap centrifuge tubes
Procedure
1. Extraction of Soluble Sugars
1.1 Weigh 0.5–1 g of fresh plant sample (dry sample is also acceptable), cut into small pieces, add about 3 mL of distilled water for homogenization, and transfer to a graduated tube. Rinse the homogenizer 2–3 times with 12 mL of distilled water and transfer the rinsate to the same container.
1.2 Seal the opening with plastic film and extract in a boiling water bath for 30 minutes. After cooling, filter the extract and transfer the filtrate to a 50 mL volumetric flask.
1.3 Collect the residue, repeat homogenization and extraction with water, combine all filtrates, and adjust to volume.
2. Dilution of Sucrose Standard
Take 1 mL of Sucrose Standard Solution (10 mg/mL) and add to a 100 mL volumetric flask. Adjust to volume with distilled water to obtain the Sucrose Standard (100 µg/mL). Using clean centrifuge tubes or test tubes, prepare a series of sucrose standards with different masses according to the table below.
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3. Sample Loading
Take 10 mL screw-cap centrifuge tubes and set up blank tubes, standard tubes, and test tubes according to the table below. Reagents should be added in the specified order, avoiding bubble formation, and mixed carefully. If the sugar concentration in the sample is too high, reduce the sample volume or dilute appropriately before measurement. It is recommended to set up 2–3 replicate tubes for each sample and calculate the average (the volumes of all reagents can be scaled down proportionally, but the minimum required amount should be maintained).
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Mix thoroughly, boil in a water bath for 1 minute, remove, and allow to cool naturally to room temperature.
4. Determination of Soluble Sugars
Mix well, set the blank tube to zero, and measure the absorbance at 630 nm for the standard tubes and test tubes using a spectrophotometer with a 1 cm light path cuvette.
5. Calculation of Results
Plot a standard curve with the mass of the series sucrose standards (No. 1–5) in µg as the x‑axis and the corresponding absorbance as the y‑axis, and determine the linear regression equation. Calculate the mass of soluble sugar in the test tube based on its absorbance, and then compute the content. Alternatively, plot the standard curve using sucrose standard concentration (µg/mL) versus absorbance and determine the soluble sugar concentration of the sample. The soluble sugar content, expressed as a mass percentage (%), is calculated as follows:
Soluble sugar content (%) = (m₁ × VT× N) / (m₀ × VS× 1,000,000) × 100%
= (c × VT× N) / (m₀ × 1,000,000) × 100%
Parameter Description:
m₁: Mass of soluble sugar obtained from the standard curve (µg)
VT: Total volume of extraction solution (mL)
N: Dilution factor of the sample extraction solution
m₀: Mass of the plant sample (g)
V<sub>S</sub>: Volume of sample extraction solution taken for measurement (mL)
c: Soluble sugar concentration of the sample (µg/mL)
1,000,000: Conversion factor between µg and g
Notes
The test solution must be clear and transparent; no protein precipitation should occur after heating. If the sample color is too dark, decolorize with activated carbon before measurement.
If the soluble sugar concentration of the sample is too high, dilute with distilled water. The optimal sugar concentration range is 10–100 µg/mL.
Concentrated sulfuric acid (specific gravity 1.84) is highly oxidizing and corrosive, posing significant hazards. Handle with extreme care. Add concentrated sulfuric acid slowly to avoid violent boiling due to heat generation, which may cause skin or clothing burns. In case of contact, rinse immediately with running water and seek medical attention if necessary.
The results of this method are influenced by sulfuric acid concentration and heating time; perform operations accurately and carefully.
Different sugars produce different color intensities with the anthrone reagent: fructose gives the deepest color, followed by glucose, while galactose and mannose produce lighter colors, and pentoses even lighter. Therefore, errors may arise when measuring sugar mixtures due to varying proportions of different sugars. This error does not occur when measuring a single sugar type.
For your safety and health, wear a lab coat and disposable gloves during operation.
Appendix
Standard Curve Preparation:
Following the instructions, measure the absorbance of the series standards using a spectrophotometer. Example data and standard curve are provided below (for reference only):
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The results indicate a good linear gradient for sucrose standards within the range of 10–100 µg/mL.
| P1507813 | Component | 50T | Storage |
| P1507813A | 蔗糖标准溶液 (10mg/mL) | 1 mL | 2-8℃ |
| P1507813B | 蒽酮试剂 | 30 mL | 2-8℃. Store in the dark. |
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 |
|---|---|---|
 ZJ26F0938857 |
分析证书 | P1507813 |
| ZJ26F0131496 |
分析证书 | P1507813 |
危险品化学品经营许可证（带存储）