- Detection method: Colorimetric method
Manual
Detection principle
Sucrose in plant tissue is hydrolyzed to glucose and fructose in boiling water bath under acidic conditions. 5-hydroxymethyl furfural was synthesized from fructose under acid condition and measure the ultraviolet absorption of 5-hydroxymethyl furfural. Glucose must be dissimilated into ketose structure and reduced to obtain 5-hydroxymethylfurfural, but the rate of isomerization of glucose to ketose is very slow. Therefore, the ultraviolet absorption of glucose is much smaller than fructose.
Performance characteristics
| Sample type |
Plant tissue |
| Sensitivity |
0.32 μmol/mL |
| Detection range |
0.32-70 μmol/mL |
| Detection method |
Colorimetric method |
| Assay type |
Quantitative |
| Assay time |
40 min |
| Precision |
Average inter-assay CV: 8.500%Average intra-assay CV: 3.400% |
| Other instruments required |
Micropipettor, Vortex mixer, 100℃ Water bath |
| Other reagents required |
PBS (0.01 M, pH 7.4) |
| Storage |
2-8℃ |
| Valid period |
12 months |
Dilution of sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.32-70 μmol/mL).
The recommended dilution factor for different samples is as follows (for reference only):
|
Sample type
|
Dilution factor
|
|
10% Green pepper tissue homogenization
|
1
|
|
10% Epipremnum aureum tissue homogenization
|
1
|
|
10% Cucumber tissue homogenization
|
1
|
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
