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Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood / Plasma – UHPLC Reagent Kit

Order No.: 52952/UHPLC, for 1000 tests
Parameters:
Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)

UHPLC separation in 3 min
Specific internal standard for each parameter
No post-column derivatisation required
One sample preparation and one run for both parameters

Assay validated according to IVDR (=> Declaration of Conformity)

main product photo

Vitamin B1 (TPP, thiamine pyrophosphate)

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Clinical relevance
This assay allows the quantitative determination of the physiologically active forms of vitamin B1, thiamine pyrophosphate, in human whole blood samples and of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood or plasma samples.

It is intended to be used for for screening and/or monitoring of vitamin B1 and/or B6 levels where indicated
• in patients with suspected vitamin B1 and/or B6 deficiency,
• in patients with suspected vitamin B1 and/or B6 excess,
• in patients under vitamin B1 and/or B6 supplementation therapy.

 

Product advantages
• UHPLC separation in 3 min
• Specific internal standard for each parameter
• No post-column derivatisation required
• One sample preparation and one run for both parameters

 

Assay characteristics
Thiamine pyrophosphate (TPP) and pyridoxal 5’-phosphate (PLP) can be determined in one run, with one sample preparation and without complex post-column derivatisation. Sample preparation is restricted to a single precipitation step and subsequent derivatisation and is possible with Pre-mixed Neutralisation Tubes (assay 52952/UHPLC) or 96 Well Filter Plates (assay 52952/UHPLC/F). Chromatographic separation is carried out with two Mobile Phases by UHPLC. The derivatised analytes are determined by fluorescence detection with wavelength switching. The use of two internal standards assures high precision and safety in quantifying the analytes.

 

Two types of kits available

Basic Kit
• Only sample preparation reagents and vials included
• No excess quantities of mobile phase and calibrators

52052-PREMIX-BK (U)HPLC Basic Kit
For 200 sample preparations
52044 Internal Standard, 10 ml (2x)
52005 Precipitation Reagent, 30 ml (2x)
52906 Pre-mixed Neutralisation Tubes, 100 pcs. (2x)
52007 Derivatisation Reagent 1, 2 x 0.3 ml (lyoph.) (2x)
52008 Derivatisation Reagent 2, 15 ml (2x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)

Reagent Kit
• Mobile phase and calibrator included
• UHPLC Column also included

52952/UHPLC Reagent Kit
For 1000 tests
52044 Internal Standard, 10 ml (10x)
52005 Precipitation Reagent, 30 ml (10x)
52906 Pre-mixed Neutralisation Tubes, 100 pcs. (10x)
52007 Derivatisation Reagent 1, 2 x 0.3 ml (lyoph.) (10x)
52008 Derivatisation Reagent 2, 15 ml (10x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (10x)
52911 Mobile Phase A (for UHPLC), 1000 ml (2x)
52922 Mobile Phase B (for UHPLC), 1000 ml (1x)
52003 Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (5x)
52210 UHPLC Column, equilibrated, with test chromatogram, 1 pc. (1x)
More Information
Method of Analysis UHPLC
Number of Tests 1000
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Detailed performance evaluation data for this assay can be found in the appendix of the instruction manual.
Lower and Upper Limit of Quantitation

TPP 5.0 – 750 µg/l (whole blood)
PLP 5.0 – 500 µg/l (whole blood)
PLP 1.4 – 500 µg/l (plasma)
Specimen Whole Blood / Plasma
Sample Preparation

The information on the sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

  • Mix 200 µl sample/calibrator/control with:
    + 100 µl Internal Standard
    + 300 µl Precipitation Reagent
    in an amber coloured Reaction Vial.
  • Shake 30 s (vortex), then centrifuge for 5 min.
  • Pipette 100 µl of the derivatisation mix into a Pre-mixed Neutralisation Tube.
  • Add 250 µl of the supernatant obtained in steps 1–3, mix briefly.
  • Incubate for 25 min at 60 °C (water bath).
  • Incubate for 10 min at +2 to +8 °C, then centrifuge for 2 min.
  • Transfer the supernatant into an amber coloured autosampler glass vial.
  • Inject 5 µl into the UHPLC system.
Run Time 3 min
Injection Volume 5 μl
Flow Rate 0.7 ml/min
Column Temperature +20 to +25 °C
Wavelengths

Start EX 320 nm, EM 415 nm,
after approx. 1.6 min switch to EX 367 nm, EM 435 nm
Additional Info Any common UHPLC system with binary pump and programmable fluorescence detector is suitable.
Parameters Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood / Plasma – UHPLC Reagent Kit (52952/UHPLC)
For 1000 tests

Order No. Reagent Kit components (Quantities in kit)
52911 Mobile Phase A (for UHPLC), 1000 ml (2x)
52922 Mobile Phase B (for UHPLC), 1000 ml (1x)
52003 Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (5x)
52044 Internal Standard, 10 ml (10x)
52005 Precipitation Reagent, 30 ml (10x)
52906 Pre-mixed Neutralisation Tubes, 100 pcs. (10x)
52007 Derivatisation Reagent 1, 2 x 0.3 ml (lyoph.) (10x)
52008 Derivatisation Reagent 2, 15 ml (10x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (10x)
52210 UHPLC Column, equilibrated, with test chromatogram, 1 pc. (1x)

 

Order No. Required components (not included in the Reagent Kit)
  Required for the analysis of vitamin B1 and vitamin B6 in whole blood:
0165 Vitamin B1/B6 Whole Blood Control, Level I, 5 x 2.0 ml (lyoph.)
0167 Vitamin B1/B6 Whole Blood Control, Level II, 5 x 2.0 ml (lyoph.)
   
  Required for the analysis of vitamin B6 in plasma:
36005 Plasma Calibration Standard, 5 x 1.0 ml (lyoph.)
0038 Vitamin B6 Plasma Control, Level I, 5 x 2.0 ml (lyoph.)
0039 Vitamin B6 Plasma Control, Level II, 5 x 2.0 ml (lyoph.)

 

Order No. General lab equipment (non-CE/IVD products)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs.
15060 Prefilter Housing, 1 pc.
15061 Prefilter, 0.5 µm, 5 pcs.
The following components are included in the kit:
  1. 52911 Mobile Phase A UHPLC vitamin B1 B6
    Mobile Phase A (for UHPLC)
    Order no.: 52911
    Vitamins B1 and B6 – UHPLC
    Details
  2. 52922 UHPLC Mobile Phase B vitamin B1 B6
    Mobile Phase B (for UHPLC)
    Order no.: 52922
    Vitamins B1 and B6 – UHPLC
    Details
  3. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  4. 52044 HPLC internal standard vitamin B1 vitamin B6 whole blood plasma
    Internal Standard
    Order no.: 52044

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  5. 52005 HPLC precipitation reagent vitamin B1 vitamin B6 whole blood plasma
    Precipitation Reagent
    Order no.: 52005

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  6. 52906 HPLC premixed neutralisation tubes vitamin B1 vitamin B6
    Pre-mixed Neutralisation Tubes
    Order no.: 52906

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  7. 52007 HPLC derivatisation reagent 1 vitamin B1 whole blood vitamin B6 whole blood plasma
    Derivatisation Reagent 1 (lyoph.)
    Order no.: 52007
    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
    Details
  8. 52008_derivatisation_reagent_2_vitamin_b1_whole_blood_vitamin_b6_whole_blood_plasma
    Derivatisation Reagent 2
    Order no.: 52008

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  9. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  10. 52210 Analytical Column UHPLC vitamin B1 B6
    UHPLC Column
    Order no.: 52210
    Vitamins B1 and B6 – UHPLC
    Details
The following products are not included in the kit but are required for the application of the method:
  1. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  2. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005

    Vitamin B6 in Serum/Plasma - HPLC/UHPLC

    Details
  3. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  1. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005

    Vitamin B6 in Serum/Plasma - HPLC/UHPLC

    Details
  2. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  1. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  2. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  1. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  2. Prefilter and housing
    Prefilter Housing
    Order no.: 15060
    Details
  3. Prefilter - 0.5 μm
    Prefilter - 0.5 μm
    Order no.: 15061
    Details
As a customer please login or register to gain full access.

Vitamin B1 (TPP, thiamine pyrophosphate)

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Clinical relevance
This assay allows the quantitative determination of the physiologically active forms of vitamin B1, thiamine pyrophosphate, in human whole blood samples and of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood or plasma samples.

It is intended to be used for for screening and/or monitoring of vitamin B1 and/or B6 levels where indicated
• in patients with suspected vitamin B1 and/or B6 deficiency,
• in patients with suspected vitamin B1 and/or B6 excess,
• in patients under vitamin B1 and/or B6 supplementation therapy.

 

Product advantages
• UHPLC separation in 3 min
• Specific internal standard for each parameter
• No post-column derivatisation required
• One sample preparation and one run for both parameters

 

Assay characteristics
Thiamine pyrophosphate (TPP) and pyridoxal 5’-phosphate (PLP) can be determined in one run, with one sample preparation and without complex post-column derivatisation. Sample preparation is restricted to a single precipitation step and subsequent derivatisation and is possible with Pre-mixed Neutralisation Tubes (assay 52952/UHPLC) or 96 Well Filter Plates (assay 52952/UHPLC/F). Chromatographic separation is carried out with two Mobile Phases by UHPLC. The derivatised analytes are determined by fluorescence detection with wavelength switching. The use of two internal standards assures high precision and safety in quantifying the analytes.

 

Two types of kits available

Basic Kit
• Only sample preparation reagents and vials included
• No excess quantities of mobile phase and calibrators

52052-PREMIX-BK (U)HPLC Basic Kit
For 200 sample preparations
52044 Internal Standard, 10 ml (2x)
52005 Precipitation Reagent, 30 ml (2x)
52906 Pre-mixed Neutralisation Tubes, 100 pcs. (2x)
52007 Derivatisation Reagent 1, 2 x 0.3 ml (lyoph.) (2x)
52008 Derivatisation Reagent 2, 15 ml (2x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)

Reagent Kit
• Mobile phase and calibrator included
• UHPLC Column also included

52952/UHPLC Reagent Kit
For 1000 tests
52044 Internal Standard, 10 ml (10x)
52005 Precipitation Reagent, 30 ml (10x)
52906 Pre-mixed Neutralisation Tubes, 100 pcs. (10x)
52007 Derivatisation Reagent 1, 2 x 0.3 ml (lyoph.) (10x)
52008 Derivatisation Reagent 2, 15 ml (10x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (10x)
52911 Mobile Phase A (for UHPLC), 1000 ml (2x)
52922 Mobile Phase B (for UHPLC), 1000 ml (1x)
52003 Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (5x)
52210 UHPLC Column, equilibrated, with test chromatogram, 1 pc. (1x)
More Information
Method of Analysis UHPLC
Number of Tests 1000
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Detailed performance evaluation data for this assay can be found in the appendix of the instruction manual.
Lower and Upper Limit of Quantitation

TPP 5.0 – 750 µg/l (whole blood)
PLP 5.0 – 500 µg/l (whole blood)
PLP 1.4 – 500 µg/l (plasma)
Specimen Whole Blood / Plasma
Sample Preparation

The information on the sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

  • Mix 200 µl sample/calibrator/control with:
    + 100 µl Internal Standard
    + 300 µl Precipitation Reagent
    in an amber coloured Reaction Vial.
  • Shake 30 s (vortex), then centrifuge for 5 min.
  • Pipette 100 µl of the derivatisation mix into a Pre-mixed Neutralisation Tube.
  • Add 250 µl of the supernatant obtained in steps 1–3, mix briefly.
  • Incubate for 25 min at 60 °C (water bath).
  • Incubate for 10 min at +2 to +8 °C, then centrifuge for 2 min.
  • Transfer the supernatant into an amber coloured autosampler glass vial.
  • Inject 5 µl into the UHPLC system.
Run Time 3 min
Injection Volume 5 μl
Flow Rate 0.7 ml/min
Column Temperature +20 to +25 °C
Wavelengths

Start EX 320 nm, EM 415 nm,
after approx. 1.6 min switch to EX 367 nm, EM 435 nm
Additional Info Any common UHPLC system with binary pump and programmable fluorescence detector is suitable.
Parameters Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood / Plasma – UHPLC Reagent Kit (52952/UHPLC)
For 1000 tests

Order No. Reagent Kit components (Quantities in kit)
52911 Mobile Phase A (for UHPLC), 1000 ml (2x)
52922 Mobile Phase B (for UHPLC), 1000 ml (1x)
52003 Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (5x)
52044 Internal Standard, 10 ml (10x)
52005 Precipitation Reagent, 30 ml (10x)
52906 Pre-mixed Neutralisation Tubes, 100 pcs. (10x)
52007 Derivatisation Reagent 1, 2 x 0.3 ml (lyoph.) (10x)
52008 Derivatisation Reagent 2, 15 ml (10x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (10x)
52210 UHPLC Column, equilibrated, with test chromatogram, 1 pc. (1x)

 

Order No. Required components (not included in the Reagent Kit)
  Required for the analysis of vitamin B1 and vitamin B6 in whole blood:
0165 Vitamin B1/B6 Whole Blood Control, Level I, 5 x 2.0 ml (lyoph.)
0167 Vitamin B1/B6 Whole Blood Control, Level II, 5 x 2.0 ml (lyoph.)
   
  Required for the analysis of vitamin B6 in plasma:
36005 Plasma Calibration Standard, 5 x 1.0 ml (lyoph.)
0038 Vitamin B6 Plasma Control, Level I, 5 x 2.0 ml (lyoph.)
0039 Vitamin B6 Plasma Control, Level II, 5 x 2.0 ml (lyoph.)

 

Order No. General lab equipment (non-CE/IVD products)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs.
15060 Prefilter Housing, 1 pc.
15061 Prefilter, 0.5 µm, 5 pcs.
The following components are included in the kit:
  1. 52911 Mobile Phase A UHPLC vitamin B1 B6
    Mobile Phase A (for UHPLC)
    Order no.: 52911
    Vitamins B1 and B6 – UHPLC
    Details
  2. 52922 UHPLC Mobile Phase B vitamin B1 B6
    Mobile Phase B (for UHPLC)
    Order no.: 52922
    Vitamins B1 and B6 – UHPLC
    Details
  3. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  4. 52044 HPLC internal standard vitamin B1 vitamin B6 whole blood plasma
    Internal Standard
    Order no.: 52044

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  5. 52005 HPLC precipitation reagent vitamin B1 vitamin B6 whole blood plasma
    Precipitation Reagent
    Order no.: 52005

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  6. 52906 HPLC premixed neutralisation tubes vitamin B1 vitamin B6
    Pre-mixed Neutralisation Tubes
    Order no.: 52906

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  7. 52007 HPLC derivatisation reagent 1 vitamin B1 whole blood vitamin B6 whole blood plasma
    Derivatisation Reagent 1 (lyoph.)
    Order no.: 52007
    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
    Details
  8. 52008_derivatisation_reagent_2_vitamin_b1_whole_blood_vitamin_b6_whole_blood_plasma
    Derivatisation Reagent 2
    Order no.: 52008

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  9. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  10. 52210 Analytical Column UHPLC vitamin B1 B6
    UHPLC Column
    Order no.: 52210
    Vitamins B1 and B6 – UHPLC
    Details
The following products are not included in the kit but are required for the application of the method:
  1. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  2. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005

    Vitamin B6 in Serum/Plasma - HPLC/UHPLC

    Details
  3. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  1. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005

    Vitamin B6 in Serum/Plasma - HPLC/UHPLC

    Details
  2. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  1. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  2. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  1. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  2. Prefilter and housing
    Prefilter Housing
    Order no.: 15060
    Details
  3. Prefilter - 0.5 μm
    Prefilter - 0.5 μm
    Order no.: 15061
    Details
As a customer please login or register to gain full access.