Vitamin C HPLC Assay Kit


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Vitamin C HPLC Assay kit:
For Research Use Only
Size:  100 tests
Incubation Time:  10 minutes
Sample Type:  Lithium Heparinate Plasma, Serum   
Sample Size: 200 µL

Reference Values (Normal):  4 – 21 mg/l
It is suggested that each laboratory establish its own normal ranges

Intended Use
The Eagle Biosciences Vitamin C HPLC Assay kit is intended for the quantitative determination of vitamin C in Li-heparinate plasma and serum.  The Vitamin C HPLC Assay kit is for research use only and not to be used in diagnostic procedures.

Assay Principle
For the determination of vitamin C a precipitation step, which removes high molecular substances is performed first. After centrifugation the supernatant is injected into the HPLC system.  The isocratic separation via HPLC at 30°C uses a “reversed phase” column. One run lasts 12 minutes. The chromatograms are recorded by a UV-detector. The quantification is performed by the delivered plasma calibrator; the concentration is calculated via integration of the peak areas, resp. peak heights.

  1. Pipette into 1.5 ml reaction tubes:  200 µl sample, CAL or CTRL + 200 µl PREC
  2. Vortex briefly. Leave the tubes for 10 minutes at 2-8°C and centrifuge at 10.000g for 10 minutes.
  3. Inject 20 µl of the supernatant into the HPLC-system
  4. The supernatant is stable in the dark for at least 24 hours at 2-8°C.

Typical Chromatogram:


Assay Background
The disease, which is called “scurvy”, was known already 1550.  The symptoms were known also by Greeks and Romans. In the 15th to 17th century more sailors died of scurvy than of any other circumstances. The food provided on board contained nearly no vitamin C containing goods.  In the 16th century the importance of vitamin C in healing scurvy was discovered.

Vitamin C is a strong reducing agent. The oxidation of vitamin C leads to dehydroascorbic acid via a radicalic intermediate, in vivo. The three forms constitute a reversible redox-system.  The most important function of ascorbic acid is the catalization in the biosynthesis of collagen. It is important for the de novo synthesis of bone, cartilage and teeth and wound healing. Vitamin C is needed for the synthesis of noradrenalin. Vitamin C plays an important role as antioxidant. It protects other molecules and cell structures from free radicals and reactive oxygen species. Ascorbic acid promotes the resorption of iron in the intestine and reduces the production of nitrosamines which might be carcinogenic. 

The Eagle Biosciences Vitamin C HPLC Assay kit makes it possible to determine the vitamin in an easy, fast and precise method. The Vitamin C HPLC Assay kit contains all reagents ready to use for preparation and separation of the samples with exception of the column (IC2900rp) and the controls (IC2900ko). Both can be supplied by Eagle Biosciences.  Beside the complete test kit it is possible to order all components separately. Please request our single component price list.

Limitations of the Assay:
EDTA-blood should not be used in the Vitamin C HPLC Assay kit.