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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)
*Metaphosphoric acid solution necessary for sample preparation is not included in this kit.

Assay protocol

  Sample ・ Control
(S)      (C)
Standard
(Std)
  Sample

Standard
200µl
Reagent[1] 12µl 12µl
Incubate 3-5 minutes at room temperature
Reagent[2]*1 70µl 70µl
 
Solution A Solution B
  Sample
(S)
Control
(C)
Standard
(Std)
  Solution A
80µl
Solution A
80µl
Solution B
80µl
Reagent[3] 20µl ---- 20µl
Incubate 3 hours at 37°C*2
85% SA*3 100µl 100µl 100µl
Reagent[3] ---- 20µl ----
Incubate 30 minutes at room temperature
Dispense 100 l 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 l 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

SML-ROIK02-EX 1KIT

¥ 30,000
$ 400
€ 300

References
  •  Bradley DW. et al,;  Vitamin C in plasma: a comparative study of the vitamin stabilized with trichloroacetic acid or metaphosphoric acid and the effects of storage at -70 degrees, -20 degrees, 4 degrees, and 25 degrees on the stabilized vitamin. Clin Chim Acta. 1973 Feb 28;44(1):47-52.  PMID: 4707637
  •  Pialoux V. et al,;  Effects of exposure to intermittent hypoxia on oxidative stress and acute hypoxic ventilatory response in humans. Am J Respir Crit Care Med. 2009 Nov 15;180(10):1002-9. doi: 10.1164/rccm.200905-0671OC. Epub 2009 Aug 27. PMID: 19713446

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.