计算溶液所需的质量、体积或浓度。
DNS试剂 (NY/T法)
有货
为什么选择此级别
BioReagent,即用型 级 ，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。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
植物体内的碳素营养状况以及农产品的品质形状，常以糖含量作为重要指标。单糖和某些寡糖(如麦芽糖)含有游离的醛基或酮基，具有还原性，属于还原糖；多糖和蔗糖等属于非还原糖。可以利用多糖能被酸水解为单糖的特性通过测定水解后的单糖含量对总糖进行测定。
DNS试剂 (NY/T法)检测原理是还原糖在碱性条件下被氧化成糖酸，3,5-二硝基水杨酸被还原为棕红色的氨基化合物 。在一定范围内，还原糖的量与棕红色产物的颜色深浅程度呈一定比例关系 ，在540 nm处测定棕红色物质的吸光度，该吸光度值与还原糖含量呈线性关系，利用比色法和标准曲线测得样品中的还原糖和总糖的含量。
本试剂仅用于科研领域，不宜用于临床诊断或其他用途。
自备材料：
1、蒸馏水、6 M盐酸溶液、6 M氢氧化钠溶液
2、50 mL离心管
3、离心机
4、水浴锅或恒温箱
5、分光光度计、比色皿
6、匀浆器或研钵
7、I₂-KI溶液：称取5 g碘和10 g碘化钾，溶于100 mL蒸馏水中。
8、葡萄糖（Glu） 标准液：1mg/mL
操作步骤(仅供参考)：
1、还原糖的提取：
①称取植物样品0.5~3 g，剪碎，加入蒸馏水约3 mL匀浆，转移至烧杯或三角瓶中，用12 mL蒸馏水冲洗研磨器2~3次，洗出液也转移至该容器。
②50℃水浴30 min，并不时搅拌， 以便还原糖彻底浸出。
③将沉淀和浸出液转移至50 mL离心管，4000 g离心5 min。
④留取上清液， 向沉淀中加入20 mL蒸馏水，混匀，再次4000 g离心5 min。
⑤留取上清液，将2次获得的上清液合并，用蒸馏水定容至100 mL (提取液)，混匀，作为还原糖待测液。
2、总糖的水解和提取：
①称取植物样品0.5~3 g，剪碎，加入蒸馏水约3 mL匀浆，转移至烧杯或三角瓶中，用12 mL蒸馏水冲洗研磨器2~3次，洗出液也转移至该容器。
②向容器中加入10 mL 6 M盐酸溶液，搅拌均匀，煮沸30 min，并不时搅拌。
③取2滴滴加于载玻片上，滴加1滴显色液（建议I 2 -KI溶液，浓度见说明书第一页自备材料项下），检查水解是否完全，如已经水解完全则不显示蓝色。
④水解完毕后，冷却至室温，加入6 M氢氧化钠溶液，使溶液 pH 至 7.4 ，用蒸馏水定容至100 mL，混匀，4000 g离心5 min或过滤。
⑤取上清或滤液10 mL，用蒸馏水定容至100 mL，成稀释10倍的总糖水解液(提取液)，取1 mL总糖水解液，测定其还原糖的含量。
3 、制作葡萄糖标准曲线：
取干净离心管或试管，按下表进行操作，以0号调零，检测540 nm处吸光度， 以吸光度值为纵坐标，各标准浓度(mg/mL)为横坐标作图得标准曲线。
注：加入 DNS试剂后，沸水浴准确煮沸5 min，取出，自来水冷却至室温。 |
4、总糖及还原糖测定：
取制备好的还原糖提取液或总糖水解液1 mL，分别加入DNS试剂2 mL，其余操作同标准曲线的操作，于540 nm处测定各管的吸光度。
计算：
还原糖的百分含量：
还原糖含量(%)=(C×VT)/(m× VS) × 100
总糖的百分含量：
总糖含量(%)=(C×VT)/(m× VS ) × 100× 10×0.9
式中：C=从标准曲线查的糖量(mg/mL)
VT=提取液的体积(mL)
m=植物样品的质量(mg)
VS =测定时用的样品体积(mL)
注意事项：
1 、上述低温试剂应避免反复冻融，并避光保存， 以免失效或效率下降。
2 、待测样本如不能及时测定，应置于 2~8℃保存，3 天内稳定。
3 、如果样品还原糖浓度过高，应用蒸馏水稀释后重测，结果乘以稀释倍数。
4 、总糖计算公式在测定干扰杂质很少、还原糖含量相对总糖含量很少时使用，×0.9 是为了从测定出的总糖水解成单糖中，扣除水解时所消耗的水量。
5 、一般市售盐酸浓度在12 M左右，应稀释至6 M使用（具体稀释倍数需根据客户自备盐酸的浓度而定，且6 M盐酸易挥发，应现配现用）。
6、试剂开封后应尽快使用，以免影响后续实验结果。
The carbon nutritional status and quality traits of plants are often assessed using sugar content as an important indicator. Monosaccharides and certain oligosaccharides (e.g., maltose) contain free aldehyde or ketone groups and possess reducing properties, thus they are classified as reducing sugars. Polysaccharides and sucrose, among others, are non-reducing sugars. Taking advantage of the fact that polysaccharides can be hydrolyzed by acid into monosaccharides, the total sugar content can be determined by measuring the monosaccharide content after hydrolysis.
The detection principle of the DNS Reagent (NY/T Method) is that reducing sugars are oxidized to sugar acids under alkaline conditions, while 3,5-dinitrosalicylic acid is reduced to a brownish-red amino compound. Within a certain range, the amount of reducing sugars is proportional to the color intensity of the brownish-red product. The absorbance of the brownish-red substance is measured at 540 nm, and this absorbance value exhibits a linear relationship with the reducing sugar content. The content of reducing sugars and total sugars in samples can be determined using colorimetry and a standard curve.
This reagent is intended for research use only and is not suitable for clinical diagnosis or other applications.
Materials Required (User-Supplied):
1.Distilled water, 6 M hydrochloric acid solution, 6 M sodium hydroxide solution
2.50 mL centrifuge tubes
3.Centrifuge
4.Water bath or constant temperature incubator
5.Spectrophotometer and cuvettes
6.Homogenizer or mortar and pestle
7.I₂-KI solution: Dissolve 5 g of iodine and 10 g of potassium iodide in 100 mL of distilled water.
8.Glucose (Glu) standard solution: 1 mg/mL
Operating Procedure (For Reference Only):
1. Extraction of Reducing Sugars:
① Weigh 0.5–3 g of plant sample, cut into small pieces, add approximately 3 mL of distilled water, and homogenize. Transfer the homogenate to a beaker or Erlenmeyer flask. Rinse the homogenizer 2–3 times with 12 mL of distilled water and combine the washings into the same container.
② Incubate in a 50°C water bath for 30 min with intermittent stirring to ensure complete extraction of reducing sugars.
③ Transfer the precipitate and extract to a 50 mL centrifuge tube and centrifuge at 4000 × g for 5 min.
④ Retain the supernatant. Add 20 mL of distilled water to the precipitate, mix well, and centrifuge again at 4000 × g for 5 min.
⑤ Retain the supernatant. Combine the two supernatants, bring the volume to 100 mL with distilled water (extraction solution), and mix well. This serves as the reducing sugar test solution.
2. Hydrolysis and Extraction of Total Sugars:
① Weigh 0.5–3 g of plant sample, cut into small pieces, add approximately 3 mL of distilled water, and homogenize. Transfer the homogenate to a beaker or Erlenmeyer flask. Rinse the homogenizer 2–3 times with 12 mL of distilled water and combine the washings into the same container.
② Add 10 mL of 6 M hydrochloric acid solution to the container, mix well, and boil for 30 min with intermittent stirring.
③ Spot 2 drops of the hydrolysate onto a glass slide, add 1 drop of indicator solution (I₂-KI solution is recommended; see concentration details under "Materials Required" on the first page of the manual), and check whether hydrolysis is complete. Complete hydrolysis is indicated by the absence of a blue color.
④ After hydrolysis is complete, cool to room temperature. Add 6 M sodium hydroxide solution to adjust the pH to 7.4. Bring the volume to 100 mL with distilled water, mix well, and centrifuge at 4000 × g for 5 min or filter.
⑤ Take 10 mL of the supernatant or filtrate and bring the volume to 100 mL with distilled water to obtain a 10-fold diluted total sugar hydrolysate (extraction solution). Take 1 mL of this total sugar hydrolysate for determination of reducing sugar content.
3. Preparation of Glucose Standard Curve:
Take clean centrifuge tubes or test tubes and proceed according to the table below. Use tube No. 0 as the blank to zero the instrument. Measure the absorbance at 540 nm. Plot the absorbance values on the y-axis against the standard concentrations (mg/mL) on the x-axis to generate the standard curve.
Note: After adding the DNS reagent, heat in a boiling water bath for exactly 5 minutes, then remove and cool to room temperature under running tap water. |
4. Determination of Total Sugars and Reducing Sugars:
Take 1 mL of the prepared reducing sugar extract or total sugar hydrolysate, add 2 mL of DNS Reagent, and proceed with the same operations as for the standard curve. Measure the absorbance of each tube at 540 nm.
Calculation:
Reducing sugar content (%):
Reducing sugar content (%) = (C × VT) / (m × VS ) × 100
Total sugar content (%):
Total sugar content (%) = (C × VT) / (m × VS) × 100 × 10 × 0.9
C = Sugar amount determined from the standard curve (mg/mL)
VT = Volume of the extraction solution (mL)
m = Mass of the plant sample (mg)
VS= Sample volume used for the assay (mL)
Precautions:
1.Avoid repeated freeze-thaw cycles of the above low-temperature reagents and store them protected from light to prevent loss of activity or efficiency.
2.If samples cannot be assayed immediately, store at 2–8°C; they are stable for 3 days.
3.If the reducing sugar concentration of the sample is too high, dilute with distilled water and re-measure, then multiply the result by the dilution factor.
4.The total sugar calculation formula is applicable when interfering impurities are minimal and the reducing sugar content is relatively low compared to the total sugar content. The factor × 0.9 is used to account for the water consumed during hydrolysis when converting total sugars into monosaccharides.
5.Commercially available hydrochloric acid is typically around 12 M and should be diluted to 6 M for use (the exact dilution factor depends on the concentration of the customer's hydrochloric acid. Note that 6 M hydrochloric acid is volatile and should be prepared fresh immediately before use).
6.The reagent should be used as soon as possible after opening to avoid affecting subsequent experimental results.
规格
规格或纯度
BioReagent, 即用型
稳定性与储存
Store at 2-8℃ long term (12 months). Store in the dark.
英文名称
DNS Reagent (NY/T Method)
储存条件
2-8°C储存,避光
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
生物试剂/缓冲液
技术文档
高级数据
化学和物理性质
敏感性
Light-sensitive
质检证书（CoA,COO,BSE/TSE 和分析图谱)
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 |
|---|---|---|
 ZJ26F0939490 |
分析证书 | D1528801 |
| ZJ26F0939491 |
分析证书 | D1528801 |
技术文档和文章
溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
Copyright © 2023至今 上海阿拉丁生化科技股份有限公司 保留所有权利。
危险品化学品经营许可证（带存储）