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 %

Gradient

Isocratic

Information

For the HPLC analysis of vitamin B1 any isocratic HPLC system with fluorescence detector is suitable.

Column temperature

ambient (~25 °C)

Flow rate

1 ml/min

Specimen

Whole Blood

Pre-analytic Treatment

Blood samples should be collected before breakfast in the morning and prior to any medication. Lightprotected and in a closed vial patient samples are stable for 24 hours at +2 to +8 °C. For longer storage periods (up to 6 months) keep patient samples frozen 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

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

Injection Volume

50 µl

Flow

1 ml/min

Additional Info

For the HPLC analysis of vitamin B1 any isocratic HPLC system with fluorescence detector is suitable.

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

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