Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

Order no.: 52052/Premix/RUO
100 Tests
  • Vitamin B1 (TPP, thiamine pyrophosphate)
  • Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Each analyte safeguarded by a specific internal standard
No post-column derivatisation required
One sample preparation and one run for both analytes
Simplified sample prep with Pre-mixed Tubes


This Chromsystems reagent kit facilitates the combined analysis of vitamin B1 in whole blood and vitamin B6 in whole blood and plasma research samples. The efficient sample preparation with Pre-mixed Tubes processes different matrices, thus whole blood research samples can easily be measured in the same sequence with plasma research samples. Vitamin molecules are derivatised during sample preparation which renders the common post-column derivatisation unnecessary. The separation takes place with a binary gradient. Both analytes are safeguarded by specifically developed internal standards. Quality controls are also available separately.


Technical Data

Method of Analysis


Number of Tests



Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Analysis Time

7 - 9 min


Whole Blood and Whole Blood/Plasma

Sample Preparation

  • In a light protected vial mix 200 μl research sample with 100 μl Internal Standard and 300 μl Precipitation Reagent, mix for at least 30 s (vortex).
  • Centrifuge for 5 min with at least 9000 x g.
  • Add 100 μl of Derivatisation Mix and 250 μl of supernatant to a labeled Pre-mixed Neutralisation Tube, mix briefly.
  • Incubate for 25 min at 60 °C (water bath).
  • Cool research sample for 10 min at +2 to +8°C, then centrifuge for 2 min with at least 9000 x g.
  • Transfer supernatant into a light protected autosampler vial, inject 25–50 μl into the HPLC system.

Sample Stability

Protect research samples from light, at +2 to +8°C stable for 1 day, for longer storage deep freeze below -18°C (maximum 2 weeks).

Additional Info

For the HPLC analysis of vitamins B1 and B6 in whole blood or plasma every common HPLC system with binary pump and programmable fluorescence detector is suitable.

Injection Volume

25-50 µl

Flow rate

1.5 - 2.3 ml/min


Gradient profile: specific binary gradients for whole blood and plasma

Column temperature

ambient (~ 25 °C)


Start EX 320 nm, EM 415 nm,
after approx. 3.8 min switch to 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.

Kit Components

Components available separately

Neutralisation Reagent

Order no.: 52006/RUO

Mobile Phase A

Order no.: 52001/RUO

Mobile Phase B

Order no.: 52022/RUO

Internal Standard

Order no.: 52044/RUO

Precipitation Reagent

Order no.: 52005/RUO

Derivatisation Reagent 1

Order no.: 52007/RUO

Derivatisation Reagent 2

Order no.: 52008/RUO

Reaction vials

Order no.: 33005/RUO


Calibrators available separately