Vitamin B2 in whole blood - HPLC

Vitamin B2 in whole blood - HPLC

Order no.: 37000/RUO
100 Tests
  • Flavin adenine dinucleotide (FAD)
  • Flavin mononucleotide (FMN)
  • Vitamin B2 (Riboflavin)

Safe results by stabilisation of FAD
Simultaneous determination of FMN
Highly stable calibrators and controls

 

Description

The present method allows the rapid and simple determination of riboflavin, flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) in whole blood using an isocratic HPLC system. Due to their native fluorescence, the analytes can be detected directly, without derivatisation. Sample preparation is restricted to one effective protein precipitation step immediately after addition of the extraction buffer and subsequent stabilisation of the analytes. A dedicated HPLC column and a mobile phase, optimised for the specific separation, permit high performance in HPLC quantification.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Flavin adenine dinucleotide (FAD), Flavin mononucleotide (FMN), Vitamin B2 (Riboflavin)

Analysis Time

approx. 9 min

Column temperature

ambient (~20 °C)

Flow rate

1.2 ml/min

Specimen

Whole Blood

Sample Preparation

  • Pipette the following solutions into an amber reaction vial (light protected) and mix immediately for 30 s (vortex): 200 μl whole blood (or reconstituted calibrator/control) + 200 μl Extraction Buffer + 400 μl Precipitation Reagent.
  • Centrifuge for 10 minutes at a minimum of 9000 x g.
  • Pipette 100 μl of Neutralisation Buffer into an amber autosampler vial (light protected), then add 500 μl supernatant and mix well (vortex).
  • Inject 50 μl of this mixture into the HPLC system.

Sample Stability

Protect research samples from light. In a closed vial the research samples are stable for up to 1 day at +2 to +8 °C. Research samples not analysed immediately within 1 day must be kept deep-frozen below -18 °C.

Injection Volume

50 µl

Additional Info

For the analysis of vitamin B2 in whole blood every isocratic system with fluorescence detector is suitable.

Wavelength

EX 465 nm

EM 525 nm

Please note

The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.

Kit Components

Components available separately

Mobile Phase


Order no.: 37011/RUO

Mobile Phase, 10 x 1000 ml


Order no.: 37022/RUO

Extraction Buffer


Order no.: 37005/RUO

Precipitation Reagent


Order no.: 37007/RUO

Neutralisation Buffer


Order no.: 37099/RUO

Reaction vials


Order no.: 33005/RUO

Calibrators

Calibrators available separately

Controls

Controls available separately

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