35000 HPLC kit vitamin B1 whole blood

Vitamin B1 in Whole Blood - HPLC

Order no.: 35000
100 Tests
  • Vitamin B1

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

Description

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.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Vitamin B1

Analysis Time

6 min

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 %

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.

Additional Info

Every isocratic HPLC system with fluorescence detector is suitable.

Injection Volume

50 µl

Flow rate

1 ml/min

Gradient

Isocratic

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.

Kit Components

Components available separately

Calibrators

Calibrators available separately

Controls

Controls available separately