Vitamin B1 in Whole Blood - HPLC

Order No.: 35000/RUO, for 100 tests
Parameters:
Vitamin B1 (TPP, thiamine pyrophosphate)

No post-column derivatisation
Easy handling
Highly stable calibration standards and controls

Vitamin B1 (TPP, thiamine pyrophosphate)

The present method allows the rapid and simple determination of thiamine diphosphate, an analogue form of vitamin B1, in an isocratic HPLC system using fluorescent detection. The usual post-column derivatisation which requires extensive equipment is replaced here by a simple pre-column derivatisation. Sample preparation is restricted to a single precipitation step after addition of the extraction buffer and subsequent derivatisation in a reaction vial. A selected HPLC column and a mobile phase optimised for this specific separation permit high performance in HPLC quantification.

More Information
Number of Tests 100
Analysis Time approx. 5 min
Sample Preparation
  • To 200 μl whole blood/calibrator/control add 100 μl Extraction Buffer.
  • Mix briefly (vortex).
  • Add 300 μl Precipitation Reagent, mix 30 s (vortex).
  • Centrifuge 5 min at 9000 x g.
  • Pipette 200 μl of the derivatisation mix into a new vial.
  • Add 100 μl of the supernatant and mix briefly.
  • Add 100 µl Neutralisation Buffer.
  • Add 100 μl Stabilisation Buffer and mix briefly.
  • Let stand for 20 min, then inject 50 μl into the HPLC system.
Sample Stability Research samples are stable for 24 hours at +2 to +8°C (light protected in a closed vial). For longer storage periods (up to 6 months) keep research samples frozen below -18°C.
Specimen Whole Blood
Injection Volume 50 µl
Flow rate 1 ml/min
Column temperature ambient (~25 °C)
Wavelength EX 367 nm
EM 435 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.
Method of Analysis HPLC
Parameter Vitamin B1 (TPP, thiamine pyrophosphate)
Additional Info For the analysis of vitamin B1 in whole blood every isocratic system with fluorescence detector is suitable.
  1. Mobile Phase
    Mobile Phase
    Order no.: 35021/RUO

    Vitamin B1 in Whole Blood - HPLC

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  2. Mobile Phase, 10 x 1000 ml
    Mobile Phase, 10 x 1000 ml
    Order no.: 35022/RUO

    Vitamin B1 in Whole Blood - HPLC

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  3. Extraction Buffer
    Extraction Buffer
    Order no.: 37003/RUO

    Vitamin B1 in Whole Blood - HPLC

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  4. Precipitation Reagent
    Precipitation Reagent
    Order no.: 37004/RUO

    Vitamin B1 in Whole Blood - HPLC

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  5. Derivatisation Reagent 1
    Derivatisation Reagent 1
    Order no.: 35005/RUO

    Vitamin B1 in Whole Blood - HPLC

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  6. Derivatisation Reagent 2
    Derivatisation Reagent 2
    Order no.: 35006/RUO

    Vitamin B1 in Whole Blood - HPLC

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  7. Stabilisation Buffer
    Stabilisation Buffer
    Order no.: 35007/RUO

    Vitamin B1 in Whole Blood - HPLC

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  8. Neutralisation Buffer
    Neutralisation Buffer
    Order no.: 35009/RUO

    Vitamin B1 in Whole Blood - HPLC

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  9. Reaction vials
    Reaction vials
    Order no.: 33005/RUO
    Amber colour, light protected
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  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. HPLC column
    HPLC column
    Order no.: 35110/RUO
    Vitamin B1 in Whole Blood - HPLC
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  2. PEEK-encased prefilter, 5 µm
    PEEK-encased prefilter, 5 µm
    Order no.: 15009/RUO
     
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  3. PEEK prefilter housing
    PEEK prefilter housing
    Order no.: 15010/RUO
     
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  4. Precolumn cartridge holder 4/10
    Precolumn cartridge holder 4/10
    Order no.: 18001/RUO
     
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  5. Precolumn cartridge 4/10
    Precolumn cartridge 4/10
    Order no.: 18035/RUO

    Vitamin B1 in Whole Blood - HPLC

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