Vitamin B6 in Whole Blood - HPLC

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

Simple handling
Cost effectiveness achieved as a result of optimised kit assembly

CE-IVD validated product ready for IVDR within timeframes and transition periods specified by the IVDR 2017/746

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Clinical relevance

Vitamin B6 comprises pyridoxine, pyridoxamine and pyridoxal, as well as their phosphate derivatives. Pyridoxal 5'-phosphate (PLP) is the active coenzyme form of vitamin B6, and is involved in nearly a hundred different enzymatic reactions. These include synthesizing and degrading amino acids, metabolising amino acids to carbohydrates and metabolising fatty acids. Vitamin B6 is also essential for the transmission of neurotransmitters such as serotonin and dopamine, as well as for the biosynthesis of certain porphyrins. The quantity of consumed proteins is one of the things that the need for vitamin B6 is dependent on.

Vitamin B6 is available in many different foods, making vitamin B6 deficiency rare. Very unbalanced nutrition can still lead to a deficiency together with other water-soluble vitamins. This can cause a number of symptoms, including growth disorders, anaemia as a result of haem synthesis disturbances, chronic inflammation of the tongue and dermatitis. Many different metabolic disorders are also caused by a vitamin B6 deficiency, such as hyperhomocysteinemia.

An overdose from natural foods is nearly impossible as a matter of practicality, and is therefore only a result of excessive supplementation. It causes photosensitivity and certain neuropathies. Tachycardia and peripheral circulatory disorders have also been described in infants.

 

Product advantages

  • Simple handling
  • Cost effectiveness achieved as a result of optimised kit assembly
  • Method for determination in plasma/serum also available (31000/S)

 

PLP can be simply and reliably determined from whole-blood samples by using this reagent kit for HPLC. Sample preparation starts with an effective protein precipitation step to release the analyte from its proteinous holoenzyme. Subsequent derivatisation produces a fluorescent PLP derivative. The chromatographic measurement is run on an isocratic HPLC system with fluorescence detector.

Chromsystems also offers additional methods using HPLC and UHPLC to achieve combined analysis of vitamin B6 with vitamin B1.

More Information
Method of Analysis HPLC
Number of Tests 100
Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Lower Limit of Quantitation 0.6 µg/l
Upper Limit of Quantification up to 250 µg/l
Intraassay CV < 1.3 %
Interassay CV < 3.7 %
Recovery 86 %
Specimen Whole Blood
Sample Preparation
  • 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 25–50 μl into the HPLC system.
Run Time < 8 min
Injection Volume 25 - 50 µl
Flow Rate 1 - 1.2 ml/min
Column Temperature ambient (~25 °C)
Gradient Isocratic
Wavelength

EX 320 nm
EM 415 nm

Additional Info For the HPLC analysis of vitamin B6 every isocratic system with fluorescence detector is suitable.
Parameters Vitamin B6 (PLP, pyridoxal 5’-phosphate)
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

Vitamin B6 comprises pyridoxine, pyridoxamine and pyridoxal, as well as their phosphate derivatives. Pyridoxal 5'-phosphate (PLP) is the active coenzyme form of vitamin B6, and is involved in nearly a hundred different enzymatic reactions. These include synthesizing and degrading amino acids, metabolising amino acids to carbohydrates and metabolising fatty acids. Vitamin B6 is also essential for the transmission of neurotransmitters such as serotonin and dopamine, as well as for the biosynthesis of certain porphyrins. The quantity of consumed proteins is one of the things that the need for vitamin B6 is dependent on.

Vitamin B6 is available in many different foods, making vitamin B6 deficiency rare. Very unbalanced nutrition can still lead to a deficiency together with other water-soluble vitamins. This can cause a number of symptoms, including growth disorders, anaemia as a result of haem synthesis disturbances, chronic inflammation of the tongue and dermatitis. Many different metabolic disorders are also caused by a vitamin B6 deficiency, such as hyperhomocysteinemia.

An overdose from natural foods is nearly impossible as a matter of practicality, and is therefore only a result of excessive supplementation. It causes photosensitivity and certain neuropathies. Tachycardia and peripheral circulatory disorders have also been described in infants.

 

Product advantages

  • Simple handling
  • Cost effectiveness achieved as a result of optimised kit assembly
  • Method for determination in plasma/serum also available (31000/S)

 

PLP can be simply and reliably determined from whole-blood samples by using this reagent kit for HPLC. Sample preparation starts with an effective protein precipitation step to release the analyte from its proteinous holoenzyme. Subsequent derivatisation produces a fluorescent PLP derivative. The chromatographic measurement is run on an isocratic HPLC system with fluorescence detector.

Chromsystems also offers additional methods using HPLC and UHPLC to achieve combined analysis of vitamin B6 with vitamin B1.

More Information
Method of Analysis HPLC
Number of Tests 100
Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Lower Limit of Quantitation 0.6 µg/l
Upper Limit of Quantification up to 250 µg/l
Intraassay CV < 1.3 %
Interassay CV < 3.7 %
Recovery 86 %
Specimen Whole Blood
Sample Preparation
  • 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 25–50 μl into the HPLC system.
Run Time < 8 min
Injection Volume 25 - 50 µl
Flow Rate 1 - 1.2 ml/min
Column Temperature ambient (~25 °C)
Gradient Isocratic
Wavelength

EX 320 nm
EM 415 nm

Additional Info For the HPLC analysis of vitamin B6 every isocratic system with fluorescence detector is suitable.
Parameters Vitamin B6 (PLP, pyridoxal 5’-phosphate)
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.