Usefull for he measurement of total vitamin C (AsA + DHAsA)

Vitamin C Assay Kit


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Background

Vitamin C (L-Ascorbic acid) is water-soluble vitamin with strong reducing action and is an important coenzyme for internal hydroxylation reactions (e.g. collagen). Vitamin C is found in both reduced form (ascorbic acid (AsA)) and oxidized form (dehydroascorbic acid (DHAsA)). This Vitamin C Assay Kit measures total vitamin C (AsA + DHAsA).
The method of this kit is a refinement of the colorimetric assay described by Daniel (1973).

SML_ROIK02EX_1.jpg

AsA in a given sample is converted to DHAsA by oxidizing agent. DHAsA is then derivatized with DNPH. Total vitamin C concentration (AsA + DHAsA) can be determined by the specific UV absorption of the DNPH derivative.


 Oxidative Stress and Aging Research Kits Flyer [PDF]

Application

Vitamin C(L-ascorbic acid) in internal organ, tissue, plasma, fruit and vegetable can be measured.

Application example
Vitamin C Assay Report (Glucose) PDF
Vitamin C Assay Report (Tomato) PDF
Vitamin C Assay Report (Lettuce) PDF

Kit component

[Sufficient for 100 assay points]
*This kit is sufficient for 100 assay points (50 samples, duplicate assay points).
1) Reagent[1]
  Oxidizing agent 1 vial (2 ml)
2) Reagent[2]
  5% Metaphosphoric acid/2% SnCl2 1 vial (10 ml)
3) Reagent[3]
  2,4-Dinitrophenylhydrazine (DNPH) 1 vial (Dissolve in 3 ml 44% sulfuric acid)
4) Reagent[4]
  5% Metaphosphoric acid 1 vial (10 ml)
5) Vitamin C Standard Stock Solution 1 vial (1 ml)

Assay protocol

  Sample E Control
(S)@     (C)
Standard
(Std)
  Sample
200 µl
Standard
200 µl
Reagent[1] 12 µl 12 µl
Incubate 3-5 minutes at room temperature
Reagent[2]*1 70 µl 70 µl
  ¥ ¥
Solution A Solution B
¥ ¥ ¥
  Sample
(S)
Control
(C)
Standard
(Std)
  Solution A
80 µl
Solution A
80 µl
Solution B
80 µl
Reagent[3] 20 µl ---- 20 µl
Incubate 3 hours at 37°C*2
85% SA*3 100 µl 100 µl 100 µl
Reagent[3] ---- 20 µl ----
Incubate 30 minutes at room temperature
Dispense 100 µl into each well of a 96 well plate.
Read absorbance at 530 nm on microplate reader.Or, read samples with spectrophotometer
(530 nm) and 100 µl cuvette.

Real absorbance is calculated as the absorbance value of the Sample minus the absorbance value of control(C).
Correction of Standards (Std) is not necessary.

Product List

Product Name Cat# Quantity Price

Vitamin C, Assay kit DataSheet ,

SML-ROIK02-EX

1 KIT ¥30,000
$400

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