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Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
The carbon nutritional status in plants and the quality characteristics of agricultural products are often assessed using sugar content as a key indicator. Monosaccharides and some oligosaccharides (such as maltose) contain free aldehyde or ketone groups, possess reducing properties, and are classified as reducing sugars. Polysaccharides and sucrose are non-reducing sugars.
Detection Principle
Under alkaline heating conditions, reducing sugars are oxidized to sugar acids, while 3,5-dinitrosalicylic acid (DNS) is reduced to a brownish-red aminocompound. Within a certain range, the amount of reducing sugar is proportionally related to the intensity of the brownish-red color. The absorbance of this brownish-red product is measured at 540 nm using a spectrophotometer. This absorbance value shows a linear relationship with the reducing sugar content. The reducing sugar contents in the sample are calculated using a standard curve. This kit is for research use only and is not suitable for clinical diagnosis or other purposes.
Applicable Samples: Plants
Component List
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Reagents, consumables and Equipments not provided
Operating Steps (For Reference Only)
1. Extraction of Reducing Sugars
1.1 Weigh 0.5–3 g of plant sample, cut into pieces, add about 3 mL of distilled water, and homogenize. Transfer to a beaker or conical flask. Rinse the homogenizer 2–3 times with 12 mL of distilled water and transfer the rinses to the same container.
1.2 Incubate in a 50°C water bath for 30 minutes, stirring occasionally to allow thorough extraction of reducing sugars.
1.3 Transfer the mixture (precipitate and extract) to a 50 mL centrifuge tube. Centrifuge at 4000 g for 5 minutes.
1.4 Collect the supernatant. Add 20 mL of distilled water to the precipitate, mix well, and centrifuge again at 4000 g for 5 minutes.
1.5 Collect the supernatant. Combine the supernatants from the two steps and dilute to 100 mL with distilled water (extraction solution). Mix well. This is the reducing sugar test solution.
2. Preparation of Glucose Standards
Using clean centrifuge tubes or test tubes, prepare a series of Glu standards according to the table below.
| Standard Working Solution | Glu Standard (1 mg/mL) (mL) | Distilled Water (mL) | Standard Glucose Concentration (mg/mL) |
| 1 | 0.1 | 0.4 | 0.2 |
| 2 | 0.2 | 0.3 | 0.4 |
| 3 | 0.3 | 0.2 | 0.6 |
| 4 | 0.4 | 0.1 | 0.8 |
| 5 | 0.5 | 0 | 1.0 |
3. Sample Addition
Take 5 mL centrifuge tubes. Set up blank, standard, and test tubes according to the table below. Add solutions in the specified order, taking care to avoid bubbles. Mix gently. 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 per sample and take the average.
| Reagent (mL) | Blank Tube | Standard Tube | Test Tube |
| Distilled Water | 0.5 | / | / |
| Series Glu Standards (1-5) | / | 0.5 | / |
| Extraction Solution | / | / | 0.5 |
| DNS Detection Solution | 1 | 1 | 1 |
Heat in a boiling water bath for exactly 5 minutes. Remove, cool to room temperature with tap water, then add 2.5 mL of distilled water to each tube.
4. Reducing Sugar Measurement
Mix well. Zero the spectrophotometer with the blank tube. Using a 1 cm pathlength cuvette, measure the absorbance of the standard and test tubes at 540 nm.
5. Result Calculation
6.1 Standard Curve Plotting
Using the Glu standards (1-5), i.e., the standard glucose concentrations (mg/mL) as the x-axis and the corresponding absorbance values as the y-axis, plot the standard curve. Determine the corresponding glucose concentration for the extraction solution based on its absorbance from the standard curve.
6.2 Content Calculation
Percentage content of reducing sugars:
Reducing sugar content in 100 g sample (g) = (c × V<sub>T</sub>) / (m × 1000) × 100 = (c × V<sub>T</sub>) / (m × 10)
Parameter Explanation:
c: Sugar concentration obtained from the standard curve (mg/mL)
V<sub>T</sub>: Total volume of the extraction solution, 100 mL
m: Mass of the plant sample (g)
Precautions
Avoid repeated freeze-thaw cycles for the low-temperature reagents mentioned above to prevent loss of efficacy or reduced efficiency.
If test samples cannot be measured immediately, store at 2–8°C. They are stable for up to 3 days.
If the reducing sugar concentration in the sample is too high, dilute with distilled water and repeat the measurement. Multiply the result by the dilution factor.
Please use reagents as soon as possible after opening to avoid affecting subsequent experimental results.
Appendix
Standard Curve Preparation: Follow the instructions to measure the absorbance of a series of standards using a spectrophotometer. The standard curve is as follows (for reference only):
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