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LCMS - Vitamin B1 & B6 Whole Blood

Vitamin B1 & B6 LC-MS reagents kit

SKU: 1020 KIT M VB1B6

Components

  • Vitamin B1 (Thiamine diphosphate, TDP)
  • Vitamin B6 (Pyridoxal-5’-phosphate, PLP)

The Vitamin B1 & B6 LC-MS Reagent Set from Diagnotix

The Vitamin B1 & B6 LC-MS reagent set from Diagnotix is one of the most optimized and widely recognized bench methods available today. Read below to discover how you can benefit from switching to the Diagnotix B1&B6 LC-MS kit. Explore how transitioning to our Vitamin B1 & B6 reagent kit can enhance your workflow, improve accuracy, and reduce costs.

Why using the Diagnotix LC-MS solution?

  • Excellent peak shape --> Increased sensitivity
  • Baseline separation --> Increased accuracy
  • Lowered TCA-concentration --> Increased LC-MS care
  • Spot on SKML proficiency testing
  • We have implemented the method across various locations and on different machinery
  • Achieve superior stability once the method is optimally implemented on your system (system-dependent, but we are here to support you)
  • Human whole blood for standards is carefully selected and optimized to minimize or exclude interfering molecules

Why Transition from HPLC to LC-MS for B1 & B6 Measurements in Whole Blood Today?

  • Win Time in Sample Preparation (4x Faster)
  • Observe Faster Run Times (2 to 2.5x Faster)
  • Say Goodbye to Toxic Solvents Containing Cyanide (Tip: Ask your HPLC kit supplier if cyanide is included!)
  • Enjoy Saving Resources, Costs and Labor Time
  • Low TCA Concentration: Gentler on Your LC-MS System

Intended use


The Diagnotix B1&B6 LC-MS Kit is specifically designed to accurately measure B1 and B6 concentrations in whole blood, providing a reliable foundation for advanced diagnostic insights.


Technical benefits


  1. Advanced Ion-Pairing Reversed-Phase LC-MS/MS Technology: Ensures exceptionally stable chromatography without the need for volatile high-pH mobile phases, derivatization steps, or inconsistent HILIC methodologies.
  2. Lowered TCA Concentration: Lowered TCA-concentration compared to standard published methods. Taking better care to your LC-MS system.
  3. Benchmark Performance: Consistently achieves perfect scores in SKML proficiency tests, recognized as the gold standard for B1 and B6 analysis in whole blood.
  4. Real-World Calibration: Uses authentic human whole blood for calibrators and controls, ensuring unmatched accuracy and reliability.
  5. Fully Automatable: Seamlessly integrates with Tecan systems for complete automation.
  6. Outstanding Validation: Backed by exceptional validation scores—refer to our technical details for comprehensive results.
  7. Cost Efficiency: Save significant resources and reduce costs compared to traditional HPLC methods and kits.

Especially interesting for:

  1. High-Throughput Laboratories: Labs frequently analyzing B1 and B6 concentrations.
  2. LC-MS Users or Adopters: Labs equipped with LC-MS systems or those considering a transition from HPLC to LC-MS. The substantial cost savings over time may justify investing in LC-MS technology.

Contact Us for More Information


Interested in learning more about our B1&B6 in whole blood by LC-MS/MS? Contact us via the contact form below. We looking forward to hear from you!


Contact form

Methodology Details for Vitamin B1 & B6 Measurement with Diagnotix Kit


Optimal Preparation with the Diagnotix Vitamin B1 & B6 Reagent Kit


For optimal preparation using the Diagnotix Vitamin B1 & B6 reagent kit, the process involves just a few straightforward steps:

1. Add Internal Standards & Vortex:
Add deuterated Vitamin B1 and deuterated Vitamin B6 as internal standards to your samples and perform deproteinization while vortexing to ensure thorough mixing.
2. Shaking Step:
After vortexing, shake the samples for 30 minutes using a shaker to ensure proper separation.
3.Centrifugation: Spin the samples for 5 minutes at a minimum of 15,000 rpm to separate the supernatant.
4. Final Step:
Transfer the supernatant into an injection vial or well plate for LC-MS analysis.

This efficient preparation method enables accurate, fast, and reliable measurements with minimal effort and time.Once these steps are completed, both vitamins can be measured simultaneously in less than 4 minutes.

This streamlined process ensures reliable results with minimal effort and cost, making it an ideal solution for high-throughput labs.


Efficient Deproteinization with Low TCA Concentration


To prevent contamination of the LC-MS system, deproteinization is performed with a low TCA concentration. Using an innovative additive, the TCA concentration is reduced to approximately 5%, compared to the typical 20% used in conventional methods. This reduces wear on the LC-MS system and ensures better longevity.

Guaranteed Accuracy with 6-Point Hemolysate Calibration from human whole blood


The accuracy of Vitamin B1 & B6 testing is ensured through a 6-point calibration using hemolysate, which closely simulates the sample matrix. This robust calibration method guarantees reliable results across a wide range of sample types.

Quality Control with 3-Level human whole blood Control


To maintain the highest standards of test quality, a 3-level control based on the same hemolysate matrix is available. This enables continuous monitoring and ensures consistent, accurate results.

Accurate Recovery of Markers in SKML Surveys


The Diagnotix Vitamin B1 & B6 kit has been proven to accurately recover the markers stated in SKML proficiency surveys, providing confidence in the method’s precision and reliability.



At a glance

  • Matrix: Whole blood
  • Technique: LC-MS/MS
  • Internal Standard: Thiamine diphosphate-D3 (TDP-D3), Pyridoxal-5’-phosphate-D3 (PLP-D3)

Included in kit (300 assays):

  • 1022a CAL M VB1B6, Calibrator Set (Calibrator 1 – 6), 6 x 2 x 250 µl
  • 1029 M VB1B6, Internal Standard D3, 3 x 6 ml
  • 1030 M VB1B6, Deproteinization Solution, 3 x 46 ml
  • 1031 M VB1B6, Mobile Phase I, 1 x 500 ml
  • 1032 M VB1B6, Mobile Phase II, 1 x 250 ml
  • 10201 M VB1B6, Autosampler Washing Solution, 1 x 1000 ml

Clinical Relevance


Water-soluble B Vitamins are important cofactors in cell metabolism. Two water-soluble vitamins with clinical relevance are Vitamins B1 and B6. Thiamine Diphosphate (TDP) is the biologically active form of Vitamin B1 and is required for various metabolic functions. Prolonged deficiency can cause Beriberi, a debilitating neurological disease. Pyridoxal 5- phosphate (PLP) is the biologically active form of Vitamin B6 and is a coenzyme for a number of transamination reactions. It plays critical roles in chronic disease and pro-inflammatory response. Additionally, both Vitamin B6 and B1 have also been linked to increased survival rates in the elderly.

Chromatograms

Technical data

Applicable for at Least: Waters TQ-Sµ, Shimadzu LC-8050, Sciex 4500
Limit of Quantification; Vitamin B1 / Vitamin B6 (µmol/l) 6.43 / 8.00
Linearity; Vitamin B1 / Vitamin B6 (µmol/l) 8000 / 3000
Accuracy Passes (<15% CV)
Simple Precision; Vitamin B1 / Vitamin B6 (CV %) 2.4 / 3.4
Complex Precision; Vitamin B1 / Vitamin B6 (CV %) 4.7 / 4.9
Ionization Electrospray
MS/MS mode Positive
Sample Preparation 50 µl Whole blood (EDTA and Heparin) → 50 µl IS → 400 µl deprot → shaker → centrifuge → inject
Preferred Column Xbridge BEH C18 XP Column 2.5 µm, 3mm x 75 mm (conditioned before use)
Analysis Time 2.5 minutes
Flow Rate 0.6 ml/min
Injection Volume 20 µl
Validation Report Available Yes

Please Note:

Information as presented freely on this website is not sufficient for proper operation of these medical devices. Please refer to the applicable instructions for use and all accompanying information such as warning notices. All necessary documentation can be found in the download centre section of this website.

For other LC-MS/MS systems than listed above, please contact us to discuss the possibilities.

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Contact details

De Plassen 4
9902 SE Appingedam
The Netherlands

info@diagnotix.com +31 (0)596 20 10 62

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