MassChrom® Vitamins B1 and B6 in Whole Blood, with Reaction Vials

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

Fast and easy sample preparation
Robust method with high selectivity
Short run time of 2.5 minutes
Isotopically-labelled internal standard for each analyte
With reaction vials

 

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 carbohydrate metabolism and for pyruvate decarboxylase. TPP is also significantly involved in the oxidative degradation of glucose. The determination of the TPP concentration is more significant than measurements of total thiamine, as by this method only the physiologically active form is taken into account.

Vitamin B₆ comprises pyridoxine, pyridoxamine and pyridoxal, as well as their phosphate derivatives. It is ingested with food and transferred via several enzymatic conversions into its active form, pyridoxal-5’-phosphate (PLP). PLP is involved in nearly one hundred different enzymatic reactions, among other things, as co-factor in amino acid metabolism, and in the formation of haemoglobin or neurotransmitters in the brain.

Product advantages

• Fast and easy sample preparation
• Robust method with high selectivity
• Short run time of 2.5 minutes
• Isotopically-labelled internal standard for each analyte

This Chromsystems assay enables combined determination of the active forms of vitamin B1 (thiamine pyrophosphate, TPP) and vitamin B6 (pyridoxal 5‘-phosphate, PLP) in whole blood by LC-MS/MS. Sample preparation consists of a simple protein precipitation step, with no need for derivatisation. This makes for a much faster sample preparation and high sample throughput. Analytes and potential interferences are effectively and swiftly separated within 2.5 minutes, indicating the method's high selectivity and speed. The chromatographic separation of the MassChrom^® assay is fast enough to ensure a high throughput in the laboratory, but sufficiently resolving for a clear separation of the two analytes with minimal potential matrix effects. Isotope-labelled internal standards for both parameters as well as 6PLUS1^® calibrators and MassCheck^® controls ensure accurate and precise results. The assay is available in 2 configurations either with reaction vials or with 96 deep well plates.

CHROMATOGRAM

TECHNICAL DATA

Method of Analysis

Number of Tests

Parameter

Analysis Time

Limit of quantification

3.4 μg/l TPP
2.8 μg/l PLP

Linearity

850 μg/l TPP
1000 μg/l PLP

Intraassay

Interassay

Specimen

Sample Preparation

• Pipette 40 μl sample/calibrator/MassCheck® control into reaction vial.
• Add 80 μl Internal Standard Mix, mix briefly (vortex).
• Add 350 μl Precipitation Reagent, mix for 1 min (vortex) and centrifuge for 2 min.
• Transfer supernatant into autosampler vial.
• Inject 20 μl into the LC-MS/MS system.

Injection Volume

Flow rate

Column temperature

Ionisation

MS/MS-Mode

Please note

KIT COMPONENTS

Components available separately

Mobile Phase A

Order no.: 87001

DETAILS

Mobile Phase B

Order no.: 87002

DETAILS

Internal Standard Set Vitamins B1 and B6

Order no.: 87046

DETAILS

Precipitation Reagent

Order no.: 87005

DETAILS

Rinsing Solution

Order no.: 87009

DETAILS

Reaction Vials, 2 ml

Order no.: 33008

DETAILS

ACCESSORIES

PP Screw-on caps, pierceable silicone/PTFE septa, 1.0 mm

Order no.: J0410

DETAILS

Autosampler vials, screw neck, amber glass, 1.5 ml

Order no.: J0601

DETAILS

Analytical column

Order no.: 87100

DETAILS

Tuning Mix Vitamins B1 and B6

Order no.: 87015

DETAILS

Dilution Buffer

Order no.: 87018

DETAILS

Adjustment Reagent

Order no.: 87019

DETAILS

Autosampler vials, screw neck, 15 µl inner cone, clear glass, 1.1 ml

Order no.: J0602

DETAILS

PE Screw-on caps, rubber/PTFE septa, 9 mm

Order no.: J0504

DETAILS

Micro-inserts for autosampler vials

Order no.: J0403

DETAILS

CALIBRATORS

Calibrators available separately

6PLUS1® Multilevel Whole Blood Calibrator Set Vitamins B1 and B6

Order no.: 87039

DETAILS

CONTROLS

Controls available separately

MassCheck® Vitamins B1 and B6 Whole Blood Controls

DETAILS