Total Polyphenols Quantification Assay Kit
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- Detection Method
- Colorimetric
- Detection Range
- 3.5-25 μg/mL
- Minimum Detection Limit
- 3.5 μg/mL
- Application
- Quantification (Q)
- Purpose
- The kit is designed for the rapid determination of polyphenols in various samples.
- Sample Type
- Beverages, Food
- Analytical Method
- Quantitative
- Specificity
- Specific for phenol content determination.
- Characteristics
- Phenolic Quantification Assay is based on Folin-Ciocalteu method. The FC reagent contains phosphomolybdic/ phosphotungstic acid complexes1. The method relies on the transfer of electrons in alkaline medium from phenolic compounds to form a blue chromophor
- Components
- Plate and the reagents neccesary to perform the assay.
- Material not included
- Pipettes, reaction tubes, plate reader.
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- Comment
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1 ml (microassay)/ 20μL (microplate)
- Assay Time
- 15 min
- Plate
- Uncoated
- Reagent Preparation
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Reagent A -FC reagent-needs to be diluted (1:10)
- Assay Procedure
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MICROASSAY: 1.Pipette 1 mL of each standard, unknown sample or diluted sample replicate into separate clean test tubes. Refer to the Table 1 (see booklet) as a guide for diluting the standard phenol. For the diluent, use the same buffer as in the samples. 2.Add 1 mL of Reagent A -Folin-Ciocalteu Reagent previously diluted (1:10) in deionized water- to each tube. 3.Add 1 mL of Reagent B (Alkaline Working Solution) to each tube and measure the absorbance at 700 nm immediately. 4.Measure the absorbance of these standards, blanks and unknown samples at 700 nm. MICROPLATE: 1. Prepare standards containing a range of 1 to 10 μg phenol (Gallic acid) to a volume of 200 μL. Pipette 20 μL of each standard and unknown sample or diluted sample replicate into a microplate well. Refer to the Table 2 (see booklet) as a guide for diluting the standard phenol. For the diluent, use the same buffer as in the samples. 2.To each well, add 100 μL of Reagent A -FC reagent-previously diluted (1:10) in deionized water. 3.Add 80 μL ofReagent B (Alkaline Working Solution) to each well and measure the absorbance at 700 nm on a plate reader immediately. 4.Measure the absorbance of these standards, blanks and unknown samples at 700 nm.
- Calculation of Results
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1.If the spectrophotometer or microplate reader was not zeroed with the blank, then substract the average blank value from the standard and unknown sample values. 2.Create a standard curve by plotting A700nm (y-axis) vs standard, μg (x-axis). Determine the unknown sample concentration using the standard curve. 3.Standard curve example for microplate assay procedure is shown in Figure 2 (see images). 4.Dilute the unknown samples until they reach an absorbance within the limits of the standard curve.
- Restrictions
- For Research Use only
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- Preservative
- Sodium azide
- Precaution of Use
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Storage
- RT/4 °C
- Storage Comment
- FC Reagent A: RT, FC Reagent B: RT, FC Standard: 4 °C
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