Vitamin B2 HPLC Assay


The Vitamin B2 HPLC Assay is intended for the quantitative determination of Vitamin B2 (Riboflavin) in EDTA-blood. The Vitamin B2 (Riboflavin) HPLC Assay kit is for research use only and not to be used in diagnostic procedures.

Vitamin B2 HPLC Assay

The Vitamin B2 HPLC Assay is For Research Use Only

Size: 100 tests
Sensitivity: 1.0 pg/mL
Incubation Time: 5 minutes
Sample Type: EDTA Whole Blood
Sample Size: 200 µL
Alternative Name: Riboflavin

Product Developed and Manufactured in Germany

Assay Background

Vitamin B2 is a collective term for the molecules Riboflavin, Flavin Adenine Dinucleotide (FAD) and Flavin Adenine Mononucleotide (FMN). Riboflavin belongs to the group of Flavonoids (plant pigments). It only can be synthesized by plants and microorganisms. Humans and animals ingest it via the gut and convert it to the coenzymes FAD and FMN. They are distributed within the whole body with maximum concentrations in liver, kidney and heart.

FAD and FMN are important coenzymes in protein and energy metabolism. In the central nervous system vitamin B2 is involved in the control of neurohormones and biogenic amines.  A lack of vitamin B2 leads to unspecific symptoms as fatigue and listlessness. Also, erythema, itching and gingival inflammation are recognized. The metabolic pathways of carbohydrates, fatty acids, proteins, and iron and vitamin B6 metabolism are affected. A serious lack of the B2 vitamins leads to inflammation of the mucous membrane in mouth and nose, skin disease (dermatitis), ophthalmic disorder (reduction of the acuteness of vision, cataract), neurological disorders and disturbance of the iron metabolism.

Vitamin B2 (Riboflavin) HPLC Assay kit makes it possible to determine the vitamin in an easy, fast and precise way. Vitamin B2 (Riboflavin) HPLC Assay kit includes all reagents in ready to use form for preparation and separation of the samples with exception of the columns (IC2300rp) and the controls (IC2300ko). 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 B2 HPLC Assay Kit Control Set
Vitamin B2 HPLC Assay Kit Column
Vitamin B2 Whole Blood Microtiter Plate Assay Kit

Additional Information

Assay Principle

For the determination of vitamin B2 a precipitation step to remove high molecular substances is performed first. After centrifugation the supernatant is mixed with a stabilization solution and injected into the HPLC system.  The isocratic separation via HPLC at 30°C lasts 10 minutes. The chromatograms are recorded by a fluorescence detector. The quantification is performed with the delivered whole blood calibrator; the concentration is calculated via integration of the peak heights respectively areas.

Assay Procedure

  1. Pipette into 1.5 ml reaction tubes:  300 µl PREC.  +  200 µl sample, CAL or CTRL (The precipitation reagent should be at 2-8 °C).
  2. Mix well (30 s on a Vortex-mixer)
  3. Add 500 µl STAB, vortex briefly and centrifuge for 5 min at 10.000 g.
  4. Inject 50 µl of the supernatant for chromatography into the HPLC-system.

Limitations of the Assay

Lipemic samples should not be measured with the Vitamin B2 (Riboflavin) HPLC Assay kit.

Package Inserts

Product Documents

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.