Ascorbic acid (Vc) test kit instructions - Huaqiang Electronic Network

Instructions for Ascorbic Acid (Vc) Test Kit

Kit Composition and Configuration (50T/48 samples)

Reagent 1: 5 ml of stock solution in one bottle. Store at 4°C for up to 6 months.

Reagent 1 Application Solution: When in use, mix reagent with double distilled water in a ratio of 1:14 and prepare as needed.

Reagent 2: Powder in one bottle. Store at 4°C for 6 months.

Reagent 2 Application Solution: Dissolve the powder in 0.36 ml of glacial acetic acid, then dilute to 40 ml with water. Store at 4°C.

Reagent 3: White crystal powder in one bottle. Store at 4°C for 6 months.

Reagent 3 Application Solution: Dissolve in Jiaonan (likely referring to a specific solvent), and after 2–4 hours, add 60 ml of 95% or anhydrous ethanol. Store at 4°C.

Reagent 4: Yellow stock solution, 0.2 ml per bottle. Store in a refrigerator between 4–8°C, away from light. Shelf life is 6 months.

Reagent 4 Application Solution: Take 0.15 ml of the stock solution and dilute it to 25 ml with water. Use immediately after preparation.

Reagent 5: Liquid, 6 ml per bottle. Store at 4°C for 6 months.

Reagent 6: Vc Standard: 6 mg of powder in three bottles. Each contains vitamin C. Store at 4°C for 6 months.

Vc Standard Application Solution (60 μg/ml): Dissolve the standard powder in 10 ml of reagent application solution.

Vc Standard Application Solution (6 μg/ml): Dilute the 60 μg/ml solution 10 times using reagent application solution for use.

Note: Vitamin C standards are prone to oxidation. It's recommended to use them immediately after preparing the sample supernatant and complete testing within 10 minutes.

Second, Procedure

Steps:

1. Preparation of Supernatant: Mix 0.15 ml of sample with 0.45 ml of Reagent 1, vortex thoroughly, incubate for 15 minutes, then centrifuge at 3500–4000 rpm for 10 minutes. The resulting liquid is the supernatant.

2. Determination of Vc in the Supernatant:

Blank Tube	Standard Tube	Measuring Tube
Reagent 1 (mL): 0.4	6 μg/ml Vc Standard (mL): 0.4	Supernatant (mL): 0.4
Reagent 2 (mL): 0.5	Reagent 2 (mL): 0.5	Reagent 2 (mL): 0.5
Reagent 3 (mL): 1	Reagent 3 (mL): 1	Reagent 3 (mL): 1
Reagent 4 (mL): 0.25	Reagent 4 (mL): 0.25	Reagent 4 (mL): 0.25
Mix well, incubate at 37°C for 30 minutes.
Reagent 5 (mL): 0.1	Reagent 5 (mL): 0.1	Reagent 5 (mL): 0.1

After mixing, let stand for 10 minutes. Measure absorbance at 636 nm with a 1 cm or 0.5 cm path length. Use double distilled water as the blank reference.