Vitamin B6 in Whole Blood – HPLC Reagent Kit

Order No.: 31000/WB, for 100 tests
Parameters:
Vitamin B6 (PLP, pyridoxal 5’-phosphate)

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

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Clinical relevance
This assay allows the quantitative determination of the physiologically active form of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood samples by HPLC.

It is intended to be used for determination of vitamin B6 levels where indicated
• in patients with suspected vitamin B6 deficiency,
• in patients with suspected vitamin B6 excess,
• in patients under vitamin B6 supplementation therapy.

 

Assay characteristics
Instead of the usual, technically demanding post-column derivatisation, simple precolumn derivatisation is used. The sample clean-up procedure includes an effective protein precipitation and extraction step to liberate PLP from its bound status. Subsequent derivatisation produces a fluorescent PLP-derivative. The chromatographic determination is run on an isocratic HPLC system with fluorescence detection.

 

Two types of kits available

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

31000-BK Basic Kit
For 100 sample preparations

31004 Precipitation Reagent, 30 ml (1x)
31005 Neutralisation Buffer, 25 ml (1x)
31006 Derivatisation Reagent, 10 ml (1x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)

Reagent Kit
• Mobile phase and calibrator included
 

31000/WB Reagent Kit
For 100 tests

31004 Precipitation Reagent, 30 ml (1x)
31005 Neutralisation Buffer, 25 ml (1x)
31006 Derivatisation Reagent, 10 ml (1x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)
31001 Mobile Phase, 1000 ml, (1x)
31003 Whole Blood Calibration Standard, 5 x 1 ml (lyoph.), (1x)
More Information
Method of Analysis HPLC
Number of Tests 100
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 1.0 – 250 µg/l
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.

  • To 200 μl of whole blood/calibrator/control add 300 μl Precipitation Reagent in a light protected vial, vortex for at least 30 s.
  • Incubate for 10 min at +4°C, then centrifuge for 5 min at 16000 x g.
  • Transfer 250 μl supernatant into new light protected vial.
  • Add 250 μl Neutralisation Reagent and 100 μl Derivatisation Reagent (contains cyanide), mix briefly.
  • Incubate for 20 min at 60°C (water bath).
  • Cool samples for 10 min at +4°C; centrifuge for 2 min at 16000 g.
  • Transfer supernatant into light protected autosampler vial.
  • Inject 50 µl into the HPLC system.
Run Time 8 min
Injection Volume 50 µl
Flow Rate 1.0 ml/min
Column Temperature +20 to +25 °C
Wavelengths

EX 320 nm
EM 415 nm

Additional Info Every isocratic HPLC system with fluorescence detector is suitable.
Parameters Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Vitamin B6 in Whole Blood – HPLC Reagent Kit (31000/WB)
For 100 tests

Order No. Reagent Kit components (Quantities in kit)
31001 Mobile Phase, 1000 ml, (1x)
31003 Whole Blood Calibration Standard, 5 x 1 ml (lyoph.), (1x)
31004 Precipitation Reagent, 30 ml (1x)
31005 Neutralisation Buffer, 25 ml (1x)
31006 Derivatisation Reagent, 10 ml (1x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)

 

Order No. Required components (not included in the Reagent Kit)
31110 HPLC Column, equilibrated, with test chromatogram, 1 pc.
0023 Vitamin B6 Whole Blood Control, Level I (lyoph.), 5 x 2 ml
0024 Vitamin B6 Whole Blood Control, Level II (lyoph.), 5 x 2 ml

 

Order No. Lab equipment
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs.
15010 PEEK Prefilter Housing, 1 pc.
15011 PEEK-encased Prefilter, 2 µm, 5 pcs.
18001 Precolumn Cartridge Holder 4/10, 1 pc.
18010 Precolumn Cartridge 4/10, 1 pc.
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:
As a customer please login or register to gain full access.

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Clinical relevance
This assay allows the quantitative determination of the physiologically active form of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood samples by HPLC.

It is intended to be used for determination of vitamin B6 levels where indicated
• in patients with suspected vitamin B6 deficiency,
• in patients with suspected vitamin B6 excess,
• in patients under vitamin B6 supplementation therapy.

 

Assay characteristics
Instead of the usual, technically demanding post-column derivatisation, simple precolumn derivatisation is used. The sample clean-up procedure includes an effective protein precipitation and extraction step to liberate PLP from its bound status. Subsequent derivatisation produces a fluorescent PLP-derivative. The chromatographic determination is run on an isocratic HPLC system with fluorescence detection.

 

Two types of kits available

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

31000-BK Basic Kit
For 100 sample preparations

31004 Precipitation Reagent, 30 ml (1x)
31005 Neutralisation Buffer, 25 ml (1x)
31006 Derivatisation Reagent, 10 ml (1x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)

Reagent Kit
• Mobile phase and calibrator included
 

31000/WB Reagent Kit
For 100 tests

31004 Precipitation Reagent, 30 ml (1x)
31005 Neutralisation Buffer, 25 ml (1x)
31006 Derivatisation Reagent, 10 ml (1x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)
31001 Mobile Phase, 1000 ml, (1x)
31003 Whole Blood Calibration Standard, 5 x 1 ml (lyoph.), (1x)
More Information
Method of Analysis HPLC
Number of Tests 100
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 1.0 – 250 µg/l
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.

  • To 200 μl of whole blood/calibrator/control add 300 μl Precipitation Reagent in a light protected vial, vortex for at least 30 s.
  • Incubate for 10 min at +4°C, then centrifuge for 5 min at 16000 x g.
  • Transfer 250 μl supernatant into new light protected vial.
  • Add 250 μl Neutralisation Reagent and 100 μl Derivatisation Reagent (contains cyanide), mix briefly.
  • Incubate for 20 min at 60°C (water bath).
  • Cool samples for 10 min at +4°C; centrifuge for 2 min at 16000 g.
  • Transfer supernatant into light protected autosampler vial.
  • Inject 50 µl into the HPLC system.
Run Time 8 min
Injection Volume 50 µl
Flow Rate 1.0 ml/min
Column Temperature +20 to +25 °C
Wavelengths

EX 320 nm
EM 415 nm

Additional Info Every isocratic HPLC system with fluorescence detector is suitable.
Parameters Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Vitamin B6 in Whole Blood – HPLC Reagent Kit (31000/WB)
For 100 tests

Order No. Reagent Kit components (Quantities in kit)
31001 Mobile Phase, 1000 ml, (1x)
31003 Whole Blood Calibration Standard, 5 x 1 ml (lyoph.), (1x)
31004 Precipitation Reagent, 30 ml (1x)
31005 Neutralisation Buffer, 25 ml (1x)
31006 Derivatisation Reagent, 10 ml (1x)
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x)

 

Order No. Required components (not included in the Reagent Kit)
31110 HPLC Column, equilibrated, with test chromatogram, 1 pc.
0023 Vitamin B6 Whole Blood Control, Level I (lyoph.), 5 x 2 ml
0024 Vitamin B6 Whole Blood Control, Level II (lyoph.), 5 x 2 ml

 

Order No. Lab equipment
33005 Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs.
15010 PEEK Prefilter Housing, 1 pc.
15011 PEEK-encased Prefilter, 2 µm, 5 pcs.
18001 Precolumn Cartridge Holder 4/10, 1 pc.
18010 Precolumn Cartridge 4/10, 1 pc.
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:
As a customer please login or register to gain full access.