Plant Flavonoids Colorimetric Assay Kit
SKU: E-BC-K284-M-500
Plant Flavonoids Colorimetric Assay Kit
SKU # | E-BC-K284-M |
Detection Instrument | Microplate reader (500 nm-520 nm, optimum wavelength: 510 nm) |
Detection method | Colorimetric method |
Product Details
Properties
Sample type | Plant tissue |
Sensitivity | 0.66 μg/mL |
Detection range | 0.66-150 μg/mL |
Detection Method | Colorimetric method |
Assay type | Quantitative |
Assay time | 35 min |
Precision | Average inter-assay CV: 5.300% | Average intra-assay CV: 4% |
Other instruments required | Micropipettor, Vortex mixer, Centrifuge, Vacuum dryer, Ultrasonic cell disruptor. |
Other reagents required |
60% alcohol |
Storage | 2-8℃ |
Valid period | 12 months |
Images
JH Kim et al investigate the bisphenol A-induced inflammation and cytotoxicity blocked by Grifola frondosa extract. Plant flavonoids level in plant tissue was determined using plant flavonoids colorimetric assay kit (E-BC-K284-M).
The concentration of total flavonoid was determined.
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 (0.66-150 μg/mL).
The recommended dilution factor for different samples is as follows (for reference only):
Sample type | Dilution factor |
Green dill | 10-15 |
Green pepper | 1 |
Pumpkin | 1 |
Heather | 25-35 |
Note: The diluent is 60% alcohol.
Detection Principle
In alkaline nitrite solution, flavonoids form red complex with aluminum ion. The flavonoid content of the sample can be calculated by measuring the absorbance of the sample extract at 510 nm.
Kit Components & Storage
Item | Component | Size 1 (48 T) | Size 2 (96 T) | Storage |
Reagent 1 | 1 mg/mL Standard | 1.8 mL × 1 vial | 1.8 mL × 1 vial | 2-8°C, 12 months |
Reagent 2 | Saline Solution | 1 mL × 1 vial | 1 mL × 2 vials | 2-8°C, 12 months |
Reagent 3 | Aluminium Reagent | 1.8 mL × 1 vial | 1.8 mL × 2 vials | 2-8°C, 12 months |
Reagent 4 | Alkali Reagent | 15 mL × 1 vial | 30 mL × 1 vial | 2-8℃, 12 months |
Microplate | 96 wells | No requirement | ||
Plate Sealer | 2 pieces |
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)
Parameters | Sample 1 | Sample 2 | Sample 3 |
Mean (μg/mL) | 2.60 | 34.50 | 85.40 |
%CV | 4.5 | 4.2 | 3.3 |
Inter-assay Precision
Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.
Parameters | Sample 1 | Sample 2 | Sample 3 |
Mean (μg/mL) | 2.60 | 34.50 | 85.40 |
%CV | 5.6 | 4.8 | 5.5 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 103%.
Parameters | Standard 1 | Standard 2 | Standard 3 |
Expected Conc. (μg/mL) | 26 | 75 | 116 |
Observed Conc. (μg/mL) | 27.3 | 77.3 | 117.2 |
Recovery rate (%) | 105 | 103 | 101 |
Sensitivity
The analytical sensitivity of the assay is 0.66 μg/mL. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve
As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
Concentration (μg/mL) | 0 | 20 | 40 | 60 | 80 | 100 | 120 | 150 |
OD value | 0.041 | 0.082 | 0.132 | 0.181 | 0.235 | 0.272 | 0.338 | 0.405 |
0.040 | 0.083 | 0.134 | 0.183 | 0.233 | 0.281 | 0.340 | 0.413 | |
Average OD | 0.041 | 0.082 | 0.133 | 0.182 | 0.234 | 0.277 | 0.339 | 0.409 |
Absoluted OD | 0.000 | 0.042 | 0.093 | 0.142 | 0.194 | 0.237 | 0.299 | 0.369 |