Vitamin B1 in Whole Blood - HPLC

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

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

CE-IVD validated product ready for IVDR within timeframes and transition periods specified by the IVDR-2022 regulation

Vitamin B1 (TPP, thiamine pyrophosphate)

Clinical relevance

Vitamin B1 (thiamine) plays an important role in deriving energy from carbohydrates and fats. The active form is thiamine pyrophosphate (TPP), a coenzyme for some enzymatic reactions in the carbohydrate metabolism and for the pyruvate decarboxylase. TPP is also significantly involved in the oxidative degradation of glucose. The determination of TPP is more significant than measurements of total thiamine, as by this method only the physiologically active form is taken into account.

Thiamine need is closely associated with caloric intake due to its central role in the energy metabolism: the more energy is needed, e.g. for competitive sports, the more thiamine the food must contain. It also prevents the accumulation of toxic by-products of cell metabolism that may damage the heart or nervous system.

 

Product advantages

  • No post-column derivatisation
  • Simple handling
  • Stable standards and controls

 

TPP can be determined fast and simply direct from whole blood using this Chromsystems kit. The kit is optimised for isocratic HPLC systems with fluorescence detector. The derivatisation necessary for fluorescence detection is included in the sample preparation procedure, and there is no need for instrument-intensive post-column derivatisation for the fluorescence detection. A special HPLC column and an optimised mobile phase allow precise and reliable quantification in less than 6 minutes.

More Information
Method of Analysis HPLC
Number of Tests 100
Limit of quantification 0.8 μg/l
Linearity up to at least 750 μg/l
Recovery 91 %
Intraassay

CV = 2.8–4.9 %

Interassay

CV = 4.3–4.7 %

Analysis Time 6 min
Specimen Whole Blood
Pre-analytic Treatment

The patient samples are stable at +2 to +8 °C for 24 hours, light-protected in a closed vial. For longer storage (up to 6 months) keep samples below -18 °C.

Sample Preparation
  • Pipette into an amber coloured reaction vial 200 μl whole blood/calibrator/control + 100 μl Extraction Buffer.

  • Mix 2 s (vortex).

  • Add 300 μl Precipitation Reagent, mix 30 s (vortex).

  • Centrifuge 5 min at 9000 x g.

  • Pipette 200 μl derivatisation mix in a new reaction 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 Prepared samples are stable at +2 to +8 °C for at least 3 days.
Injection Volume 50 µl
Flow rate 1 ml/min
Column temperature ambient (~ 25 °C)
Wavelength

EX 367 nm

EM 435 nm

Gradient Isocratic
Additional Info

Every isocratic HPLC 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 Vitamin B1 (TPP, thiamine pyrophosphate)
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