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Vitamins B1 and B6 in Whole Blood, with 96 Well Filter Plates – UHPLC Reagent Kit

Order No.: 52952/UHPLC/F, for 960 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, and of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood 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.

Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.

 

Two types of kits available

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

52052-F-BK (U)HPLC Basic Kit
For 960 sample preparations
52744 Internal Standard Mix, 50 ml (1x)
52705 Extraction Reagent, 75 ml (2x)
52706 Prep Solution, 25 ml (3x)
52707 Finisher 1, 5 x 0.25 ml (lyoph.) (2x)
52708 Finisher 2, 50 ml (1x)
52709 Dilution Buffer, 100 ml (1x)
52057 96 Well Filter Plates, 5 pcs. (2x)
52058 Collection Plates, 10 pcs. (2x)
52059 Pierceable Adhesive Seals, 5 pcs. (2x)

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

52752/UHPLC/F Reagent Kit
For 960 tests
52744 Internal Standard Mix, 50 ml (1x)
52705 Extraction Reagent, 75 ml (2x)
52706 Prep Solution, 25 ml (3x)
52707 Finisher 1, 5 x 0.25 ml (lyoph.) (2x)
52708 Finisher 2, 50 ml (1x)
52709 Dilution Buffer, 100 ml (1x)
52057 96 Well Filter Plates, 5 pcs. (2x)
52058 Collection Plates, 10 pcs. (2x)
52059 Pierceable Adhesive Seals, 5 pcs. (2x)
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 960
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 can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

TPP 2.9 – 750 µg/l (whole blood)
PLP 1.2 – 500 µg/l (whole blood)
Specimen Whole Blood
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.

  • Pipette 100 µl whole blood (calibrator, control, samples) into a 96 Well Collection Plate.
  • Add 50 µl Internal Standard Mix.
  • Add 150 µl Extraction Reagent.
  • Mix 4 min, then separate precipitate by centrifugation (3 min).
  • Transfer 75 µl of the supernatant into a 96 Well Filter Plate.
  • Add 75 µl Prep Solution.
  • Add 50 µl finisher solution.
  • Mix 30 s.
  • Put 96 Well Filter Plate onto a new 96 Well Collection Plate, then separate precipitate by centrifugation (2 min), discard 96 Well Filter Plate.
  • Incubate at +60 °C for 45 min.
  • Add 100 µl Dilution Buffer, mix 180 s.
  • Seal the plate with an adhesive seal and transfer to the autosampler.
  • Inject 5 µl eluate 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 and Vitamin B6 in Whole Blood – UHPLC Reagent Kit (52952/UHPLC/F) – with 96 Well Filter Plates
For 960 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)
52744 Internal Standard Mix, 50 ml (1x)
52705 Extraction Reagent, 75 ml (2x)
52706 Prep Solution, 25 ml (3x)
52707 Finisher 1, 5 x 0.25 ml (lyoph.) (2x)
52708 Finisher 2, 50 ml (1x)
52709 Dilution Buffer, 100 ml (1x)
52057 96 Well Filter Plates, 5 pcs. (2x)
52058 Collection Plates, 10 pcs. (2x)
52059 Pierceable Adhesive Seals, 5 pcs. (2x)
52210 UHPLC Column, equilibrated, with test chromatogram, 1 pc. (1x)

 

Order No. Required components (not included in the Reagent Kit)
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.)

 

Order No. General lab equipment (non-CE/IVD products)
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-F Mobile Phase 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. 52744 Internal Standard Mix -Vitamin B1 B6
    Internal Standard Mix
    Order no.: 52744
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  5. 52705 Extraction Reagent-Vitamin B1 B6
    Extraction Reagent
    Order no.: 52705
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  6. Prep Solution
    Prep Solution
    Order no.: 52706
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  7. 52707 Finisher 1 - Vitamin B1 B6
    Finisher 1
    Order no.: 52707
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  8. 52708 Finisher 2 -Vitamin B1 B6
    Finisher 2
    Order no.: 52708
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  9. Dilution Buffer
    Dilution Buffer
    Order no.: 52709
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  10. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    96 Well Filter Plates
    Order no.: 52057
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  11. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    Collection Plates
    Order no.: 52058
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  12. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    Pierceable Adhesive Seals, for 96 well plates
    Order no.: 52059
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  13. 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
  1. 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
  1. Prefilter and housing
    Prefilter Housing
    Order no.: 15060
    Details
  2. 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, and of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood 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.

Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.

 

Two types of kits available

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

52052-F-BK (U)HPLC Basic Kit
For 960 sample preparations
52744 Internal Standard Mix, 50 ml (1x)
52705 Extraction Reagent, 75 ml (2x)
52706 Prep Solution, 25 ml (3x)
52707 Finisher 1, 5 x 0.25 ml (lyoph.) (2x)
52708 Finisher 2, 50 ml (1x)
52709 Dilution Buffer, 100 ml (1x)
52057 96 Well Filter Plates, 5 pcs. (2x)
52058 Collection Plates, 10 pcs. (2x)
52059 Pierceable Adhesive Seals, 5 pcs. (2x)

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

52752/UHPLC/F Reagent Kit
For 960 tests
52744 Internal Standard Mix, 50 ml (1x)
52705 Extraction Reagent, 75 ml (2x)
52706 Prep Solution, 25 ml (3x)
52707 Finisher 1, 5 x 0.25 ml (lyoph.) (2x)
52708 Finisher 2, 50 ml (1x)
52709 Dilution Buffer, 100 ml (1x)
52057 96 Well Filter Plates, 5 pcs. (2x)
52058 Collection Plates, 10 pcs. (2x)
52059 Pierceable Adhesive Seals, 5 pcs. (2x)
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 960
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 can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

TPP 2.9 – 750 µg/l (whole blood)
PLP 1.2 – 500 µg/l (whole blood)
Specimen Whole Blood
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.

  • Pipette 100 µl whole blood (calibrator, control, samples) into a 96 Well Collection Plate.
  • Add 50 µl Internal Standard Mix.
  • Add 150 µl Extraction Reagent.
  • Mix 4 min, then separate precipitate by centrifugation (3 min).
  • Transfer 75 µl of the supernatant into a 96 Well Filter Plate.
  • Add 75 µl Prep Solution.
  • Add 50 µl finisher solution.
  • Mix 30 s.
  • Put 96 Well Filter Plate onto a new 96 Well Collection Plate, then separate precipitate by centrifugation (2 min), discard 96 Well Filter Plate.
  • Incubate at +60 °C for 45 min.
  • Add 100 µl Dilution Buffer, mix 180 s.
  • Seal the plate with an adhesive seal and transfer to the autosampler.
  • Inject 5 µl eluate 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 and Vitamin B6 in Whole Blood – UHPLC Reagent Kit (52952/UHPLC/F) – with 96 Well Filter Plates
For 960 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)
52744 Internal Standard Mix, 50 ml (1x)
52705 Extraction Reagent, 75 ml (2x)
52706 Prep Solution, 25 ml (3x)
52707 Finisher 1, 5 x 0.25 ml (lyoph.) (2x)
52708 Finisher 2, 50 ml (1x)
52709 Dilution Buffer, 100 ml (1x)
52057 96 Well Filter Plates, 5 pcs. (2x)
52058 Collection Plates, 10 pcs. (2x)
52059 Pierceable Adhesive Seals, 5 pcs. (2x)
52210 UHPLC Column, equilibrated, with test chromatogram, 1 pc. (1x)

 

Order No. Required components (not included in the Reagent Kit)
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.)

 

Order No. General lab equipment (non-CE/IVD products)
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-F Mobile Phase 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. 52744 Internal Standard Mix -Vitamin B1 B6
    Internal Standard Mix
    Order no.: 52744
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  5. 52705 Extraction Reagent-Vitamin B1 B6
    Extraction Reagent
    Order no.: 52705
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  6. Prep Solution
    Prep Solution
    Order no.: 52706
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  7. 52707 Finisher 1 - Vitamin B1 B6
    Finisher 1
    Order no.: 52707
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  8. 52708 Finisher 2 -Vitamin B1 B6
    Finisher 2
    Order no.: 52708
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  9. Dilution Buffer
    Dilution Buffer
    Order no.: 52709
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  10. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    96 Well Filter Plates
    Order no.: 52057
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  11. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    Collection Plates
    Order no.: 52058
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  12. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    Pierceable Adhesive Seals, for 96 well plates
    Order no.: 52059
    Vitamin B1/B6 in Whole Blood, with 96 Well Filter Plates – HPLC/UHPLC
    Details
  13. 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
  1. 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
  1. Prefilter and housing
    Prefilter Housing
    Order no.: 15060
    Details
  2. Prefilter - 0.5 μm
    Prefilter - 0.5 μm
    Order no.: 15061
    Details
As a customer please login or register to gain full access.