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

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

Order no.: 52952/UHPLC/RUO
1000 Tests
  • Vitamin B1 (TPP, thiamine pyrophosphate)
  • Vitamin B6 (PLP, pyridoxal 5’-phosphate)

UHPLC separation in less than 3 min
Each analyte is safeguarded by a specific internal standard


The Chromsystems UHPLC kit for the combined analysis of vitamin B1 in whole blood and vitamin B6 in whole blood or plasma research samples combines the features of the proven HPLC method with the distinct advantages of UHPLC technology. The efficient sample preparation with Pre-mixed Tubes involves only one precipitation step and subsequent derivatisation. Analyte separation takes approximately 3 minutes. Both analytes are safeguarded by specifically developed internal standards. Quality controls are also available separately.

Kit content

Mobile Phase A, 2 x 1000 ml
Mobile Phase B, 1000 ml
Whole Blood Calibration Standard (lyoph.), 5 x 5 x 1 ml
Internal Standard, 10 x 10 ml
Precipitation Reagent, 10 x 30 ml
Pre-mixed Neutralisation Tubes, 10 x 100 pcs.
Derivatisation Reagent 1 (lyoph.), 10 x 2 x 0.3 ml
Derivatisation Reagent 2 , 10 x 15 ml
Reaction vials, amber coloured (light protection), 10 x 100 pcs.
UHPLC column, 1 pc.


Technical Data

Method of Analysis


Number of Tests



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

Analysis Time

3 min


Whole Blood and Whole Blood/Plasma

Sample Preparation

  • Add in a light protected vial to 200 μl research sample, 100 μl Internal Standard and 300 μl Precipitation Reagent, mix 30 s (vortex).
  • Centrifuge 5 min with at least 9000 x g.
  • Add 100 μl Derivatisation Mix in a labelled Pre-mixed Neutralisation Tube and add 250 μl of the supernatant from step 1, mix briefly.
    Do not centrifuge the precipitate!
  • Incubate 25 min at 60 °C (water bath).
  • Cool down for 10 min at +2 to +8 °C and centrifuge for 2 min with at least 9000 x g.
  • Transfer the supernatant into a light protected autosampler vial, inject 2.5–10 μl into the UHPLC 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 combined UHPLC analysis of vitamins B1 and B6 in whole blood and plasma, any major UHPLC system with binary pump and programmable fluorescence detector is suitable.

Injection Volume

2.5–10 μl

Flow rate

0.7 ml/min

Column temperature

ambient (~ 25°C)


Start: EX 320 nm, EM 415 nm
After approx. 1.6 min switch to EX 367 nm, EM 435 nm


< 600 bar

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

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