Vitamins A & E HPLC Assay


The Vitamins A & E HPLC Assay is intended for the quantitative determination of vitamin A and vitamin E in plasma and serum. The Vitamin A & Vitamin E HPLC Assay kit is for research use only and not to be used in diagnostic procedures.

Vitamins A & E HPLC Assay

The Vitamins A & E HPLC Assay is For Research Use Only

Size: 100 tests
Incubation Time: 30 minutes
Sample Type: Serum, Plasma
Sample Size: 250 µL
Reference Values (Normal):

  • Vitamin A: 316-820 µg/l
  • Vitamin E: 6.6 – 14.3 mg/l

Product Developed and Manufactured in Germany
Product Support in the USA

It is suggested that each laboratory establish its own normal ranges

Assay Background

Vitamin A and E are fat soluble vitamins. Both are stored in the body, thus overdosing is possible.  Vitamin A plays an important role in the visual process. It is mainly taken up as pre-vitamin A by nutrition. In the liver it is processed to vitamin A. Retinal is involved in the visual process as prosthetic group of the rhodopsin molecule. Beside this, vitamin A (Retinol) is important for the growth of children and the growth of skin and mucous membranes. As an antioxidant, retinol protects the body against free radicals and reactive oxygen species. Overdosing leads to headache, vertigo, sickness and vomit. Disturbance of the nervous system, skin disease and loss of hair have also been reported.

Vitamin E protects the fatty acids against oxidation. It captures free radicals and reactive oxygen species. A lack of vitamin E is recognized in lipid metabolism disorders, liver disease and in early born children. It leads to muscle dystrophia, anemia and disturbances in the nervous system.

The Eagle Biosciences Vitamin A & Vitamin E HPLC Assay kit makes it possible to determine both vitamins in an easy, fast and precise method. The Eagle Biosciences Vitamin A & Vitamin E HPLC Assay kit includes all reagents ready to use for preparation and separation of the samples with exception of the columns (IC1600rp) and the controls (IC1600ko). Both can be supplied by Eagle Biosciences.  Beside the complete test kits it is possible to order all components separately. Please request our single component price list.

Related Products

Vitamin A & Vitamin E HPLC Column
Vitamin A & Vitamin E HPLC Control Set
Vitamin C HPLC Assay Kit

Additional Information

Assay Principle

For the determination of vitamin A and E, an internal standard and the precipitation reagent are added. During the precipitation step, high molecular substances are removed. 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 15 minutes. The chromatograms are recorded by a UV-detector by two different wavelength (Vitamin A at 325 nm, Vitamin E 300 nm). The quantification is performed with the delivered plasma calibrator; the concentration is calculated by the “internal standard method” via integration of the peak heights resp. peak areas.

  1. Pipette into 1.5 ml reaction tubes:
    • Preparation of the standard:  250 µl STD + 50 µl IS + 250 µl DIL + 250 µl PREC
    • Preparation of samples and controls:  250 µl sample or CTRL + 50 µl IS + 500 µl PREC
  2. Vortex briefly. Leave the tubes for 30 minutes at 2-8°C and centrifuge at 10.000g for 2 minutes.
  3. Inject 100 µl of the supernatant into the HPLC-system

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