Vitamin B2 in Whole Blood - HPLC

Order No.: 37000/RUO, for 100 tests
Parameters:
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

 

Flavin adenine dinucleotide (FAD)

Flavin mononucleotide (FMN), Vitamin B2 (Riboflavin)

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.

More Information
Method of Analysis HPLC
Number of Tests 100
Analysis Time approx. 9 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 research sample/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 Light protected the prepared research samples are stable up to 24 hours at room temperature and up to 7 days at +2 to +8 °C. For longer storage (maximum 2 weeks) the prepared samples have to be stored below -18 °C.
Injection Volume 50 µl
Flow rate 1.2 ml/min
Column temperature ambient (~20 °C)
Wavelength

EX 465 nm

EM 525 nm

Additional Info For the analysis of vitamin B2 in whole blood every isocratic system with fluorescence detector is suitable.
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.
Parameter Flavin adenine dinucleotide (FAD), Flavin mononucleotide (FMN), Vitamin B2 (Riboflavin)
  1. Mobile Phase
    Mobile Phase
    Order no.: 37011/RUO

    Vitamin B2 in Whole Blood - HPLC

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  2. Mobile Phase, 10 x 1000 ml
    Mobile Phase, 10 x 1000 ml
    Order no.: 37022/RUO

    Vitamin B2 in Whole Blood - HPLC

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  3. Extraction Buffer
    Extraction Buffer
    Order no.: 37005/RUO

    Vitamin B2 in Whole Blood - HPLC

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  4. Precipitation Reagent
    Precipitation Reagent
    Order no.: 37007/RUO

    Vitamin B2 in Whole Blood - HPLC

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  5. Neutralisation Buffer
    Neutralisation Buffer
    Order no.: 37099/RUO

    Vitamin B2 in Whole Blood - HPLC

    Details
  6. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  1. 0033 0034 0035 HPLC vitamin B1 vitamin B2 whole blood controls
    Vitamin B1/B2 Whole Blood Controls
    Order no.: 0033/0034/0035
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  1. 37110 HPLC column vitamin B2 whole blood
    HPLC Column Vitamin B2 in Whole Blood
    Order no.: 37110
    Vitamin B2 in Whole Blood - HPLC
    Details
  1. 37110 HPLC column vitamin B2 whole blood
    HPLC Column Vitamin B2 in Whole Blood
    Order no.: 37110
    Vitamin B2 in Whole Blood - HPLC
    Details
  2. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
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  3. PEEK-encased Prefilter, 5 µm
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  4. Precolumn Cartridge Holder 4/10
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  5. Precolumn Cartridge 4/10
    Precolumn Cartridge 4/10
    Order no.: 17037/RUO

     Vitamin B2 in Whole Blood - HPLC

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