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Vitamin C (VC) Colorimetric Assay Kit
SKU: E-BC-K034-S-100
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Vitamin C (VC) Colorimetric Assay Kit
| SKU # | E-BC-K034-S |
| Detection Instrument | Spectrophotometer(533 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | VC, Ascorbic acid |
| Sample Type | Serum, plasma, tissue |
| Sensitivity | 0.35 μg/mL |
| Detection Range | 0.35-20 μg/mL |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 80 min |
| Precision | Average inter-assay CV: 3.300% | Average intra-assay CV: 2.300% |
| Other instruments required | Micropipettor, Vortex mixer, Incubator, Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4), Absolute ethanol |
| 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 (0.35-20 μg/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Human plasma | 1 |
| 10% Mouse kidney tissue homogenization | 1 |
| Rat serum | 1 |
| Mouse serum | 1 |
| 10% Rat liver tissue homogenization | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
The most obvious chemical activity of VC is that reduce Fe3+ to Fe2+, then promote iron absorption in the intestine, promote the storage and utilization of iron. Fe3+ react immediately with reducing ascorbic acid to form Fe2+. then Fe2+ react with phenanthroline and the color developing reaction occurs. The content of vitamin C in sample can be determined. Measure the OD value and calculate the VC content indirectly.
Kit Components & Storage
| Item | Component | Size 1 (50 assays) | Size 2 (100 assays) | Storage |
| Reagent 1 | Extracting Solution | 10 mL × 1 vial | 10 mL × 1 vial | 2-8℃, 12 months shading light |
| Reagent 2 | Buffer Solution | 25 mL × 1 vial | 50 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent | 6 mL ×1 vial | 12 mL ×1 vial | 2-8℃, 12 months shading light |
| Reagent 4 | Ferrum Reagent | 1 mL × 1 vial | 1 mL × 1 vial | 2-8℃, 12 months shading light |
| Reagent 5 | Stop Solution | 6 mL × 1 vial | 12 mL × 1 vial | 2-8℃, 12 months |
| Reagent 6 | VC Standard | 6 mg × 3 vials | 6 mg × 3 vials | 2-8℃, 12 months shading light |
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) | 1.20 | 9.50 | 15.60 |
| %CV | 2.5 | 2.2 | 2.2 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μg/mL) | 1.20 | 9.50 | 15.60 |
| %CV | 3.6 | 2.9 | 3.4 |
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 104%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (μg/mL) | 2.5 | 8.5 | 13.5 |
| Observed Conc. (μg/mL) | 2.5 | 9.0 | 14.3 |
| Recovery rate (%) | 100 | 106 | 106 |
Sensitivity
The analytical sensitivity of the assay is 0.07 μg/mL VC. 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.