How do you make a standard curve for ascorbic acid?

Standard Curve calibration for Ascorbic acid

1. DPPH stock = 1mg/20ml (methanol)(optimized;
2. Ascorbic acid Curve.
3. Make up the volume of the above different concs to 1ml with DPPH.
4. Incubate in dark for 30mins.
5. Record the OD at 517nm.
6. Plot the curve from OD vs concentration (ascorbic acid) and calculate the linear.

How do you measure ascorbic acid?

Calculation. For standard solution Mass Ascorbic acid = Mole iodine × Volume of iodine × 176.12 = 0.00231 × 45 × 176.12 = 91.54 mg Initially, the amount of Ascorbic acid was taken for 100mg and therefore for total amount of ascorbic acid i.e. 250 mg the ratio stands out to be 2.5 (250/100).

Why ascorbic acid is used as standard?

Ascorbic acid is used as standard because of its availability in different food sources (especially citrus fruit extracts). Ascorbic acid gives better result than gallic acid,mainly gallic acid is used for determination of phenolic content as standard.

Which method is used to determine the concentration of ascorbic acid concentration?

This method determines the vitamin C concentration in a solution by a redox titration using iodine. Vitamin C, more properly called ascorbic acid, is an essential antioxidant needed by the human body (see additional notes).

How do you prepare standard for DPPH assay?

Reagent/solutions: DPPH solution – 0.3 mM in methanol (freshly prepared), Standard Ascorbic acid solution – 1 mg/ml in methanol. Sample preparation: 1 ml of PG was dried on mild heat in a water bath; the residue was taken with methanol to make 1mg/ml (PGE1) and used for the test.

How do you dissolve a DPPH?

A stock solution may be made by dissolving the DPPH in the solvent of choice. DPPH is soluble in organic solvents such as ethanol and dimethyl formamide, which should be purged with an inert gas. The solubility of DPPH in these solvents is approximately 10 mg/ml. DPPH is sparingly soluble in aqueous solutions.

How do you isolate vitamin C?

Vitamin C is water soluble, so all you need do is crush or grind the plant material and then shake it up with water. A common method is to grind the material with some clean sand in a mortar with a pestle. The material can then be filtered or centrifuged to get rid of the debris. The vitamin C will be in the water.

How do you measure the concentration of ascorbic acid?

Ascorbic acid reacts with DCPIP, changing the colour from blue to colourless. They react in a 1:1 fashion, so if a known quantity of DCPIP solution reacts with the plant tissue extract, the quantity of DCPIP used gives a direct measure of the quantity of ascorbic acid present.

How do you prepare Dichlorophenolindophenol?

Indophenol (DCPIP) solution: Dissolve approximately 0.04 g of sodium hydrogen carbonate in 200 mL of distilled water, add approximately 0.05 g DCPIP and dissolve with shaking. This is very hard to dissolve. Leave overnight and filter. Note: DCPIP is NOT a primary standard so the concentration is only approximate.

How does the DPPH assay work?

DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radical method is an antioxidant assay based on electron-transfer that produces a violet solution in ethanol (10). This free radical, stable at room temperature, is reduced in the presence of an antioxidant molecule, giving rise to colorless ethanol solution.

How do you prepare DPPH for antioxidants?

3ml of 0.4 mM DPPH solution was taken as control. Reaction mixture was prepared by taking 3ml of methanol mixed well with 0.5 mL of aqueous & alcoholic extracts and 0.3ml of 0.4 mM DPPH solution. Solution such as blank, control and reaction mixture were allowed to incubate in the dark for 30 min.

What is the control in DPPH assay?

For DPPH assay methanol is used as blank where as DPPH + methanol is used as experimental control.