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
Method of Analysis HPLC
Number of Tests 100
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.
Specimen Whole Blood
Sample Preparation
  • To 200 μl research sample/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 (light protected).
  • 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.
Run Time approx. 5 min
Injection Volume 50 µl
Flow Rate 1 ml/min
Column Temperature ambient (~25 °C)
Wavelengths EX 367 nm
EM 435 nm
Additional Info For the analysis of vitamin B1 in whole blood every isocratic system with fluorescence detector is suitable.
Parameters Vitamin B1 (TPP, thiamine pyrophosphate)
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:

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
Method of Analysis HPLC
Number of Tests 100
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.
Specimen Whole Blood
Sample Preparation
  • To 200 μl research sample/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 (light protected).
  • 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.
Run Time approx. 5 min
Injection Volume 50 µl
Flow Rate 1 ml/min
Column Temperature ambient (~25 °C)
Wavelengths EX 367 nm
EM 435 nm
Additional Info For the analysis of vitamin B1 in whole blood every isocratic system with fluorescence detector is suitable.
Parameters Vitamin B1 (TPP, thiamine pyrophosphate)
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method: