Vitamin B6 in Serum/Plasma - HPLC

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

Determination possible for whole-blood samples and plasma/serum samples
Simple handling
Cost effectiveness achieved as a result of optimised kit assembly

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

  • Determination possible for whole-blood samples and plasma/serum samples
  • Simple handling
  • Cost effectiveness achieved as a result of optimised kit assembly

 

PLP can be simply and reliably determined from plasma or serum 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
Number of Tests 100
Limit of quantification 0.5 µg/l
Linearity up to 250 µg/l
Recovery 102 %
Intraassay CV < 0.9 %
Interassay CV < 2 %
Analysis Time < 8 min
Sample Preparation
  • To 200 μl of plasma/serum/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.
Sample Stability The prepared samples can be kept for 5 days at room temperature or refrigerated (+2 to 8°C) for at least 7 days (light protected). For longer storage (up to 2 weeks) keep them below -18 °C.
Specimen Serum/Plasma
Pre-analytic Treatment Patient samples should be kept cool and light protected for transport. Storage life is one week at +2 to +8 °C in the dark, at -20 °C several months (7).
Injection Volume 25 - 50 µl
Flow rate 1 - 1.2 ml/min
Column temperature ambient (~ 25 °C)
Wavelength

EX 320 nm
EM 415 nm

Gradient Isocratic
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.
Method of Analysis HPLC
Parameter Vitamin B6 (PLP, pyridoxal 5’-phosphate)
Additional Info Every isocratic system with fluorescence detector is suitable.

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