Vitamin B6 in Serum/Plasma - HPLC

Order No.: 31000/S/RUO, for 100 tests
Parameters:
Vitamin B6 (PLP, pyridoxal 5’-phosphate)
  • Suitable for plasma and serum
  • Fast and easy sample preparation
  • Low cost per test

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

The present reagent kit allows the rapid, easy and specific determination of vitamin B6 (pyridoxal-5'-phosphate, PLP) using a simple isocratic HPLC system. Instead of the usual, technically demanding post-column derivatisation, simple precolumn derivatisation is used. Plasma and serum research specimens (after addition of EDTA or heparin as anticoagulant) are suitable for this kit. The sample clean-up procedure includes an effective protein precipitation and extraction step with optimised conditions. Subsequent derivatisation (water bath, 60 °C) produces a fluorescent PLP-derivative. The chromatographic determination is run on an isocratic HPLC system with fluorescence detection. By this sample clean up procedure highly stable extracts are achieved, allowing the analysis of large batches.

 

More Information
Number of Tests 100
Analysis Time < 8 min
Sample Preparation
  • To 200 μl of plasma/serum (calibrator, control, research specimen) add 300 μl Precipitation Reagent in a light protected vial, vortex-mix 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, mix briefly (a precipitation will form, do not centrifuge!).
  • Add 100 μl Derivatisation Reagent (contains cyanide), mix briefly.
  • Incubate for 20 min at 60°C (water bath).
  • Cool research sample down in cold water, then incubate 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 Keep research samples cool and light protected for transport. They are stable up to to 7 days at +4°C. For longer storage keep frozen below -18°C.
Specimen Plasma/Serum
Injection Volume 25–50 µl
Flow rate 1.0–1.2 ml/min
Column temperature ambient (~ 25°C)
Wavelength EX 320 nm, EM 415 nm
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)
  1. Mobile Phase
    Mobile Phase
    Order no.: 31001/RUO

    Vitamin B6 in Plasma/Serum - HPLC

    Details
  2. Mobile Phase, 10 x 1000 ml
    Mobile Phase, 10 x 1000 ml
    Order no.: 31002/RUO

    Vitamin B6 in Plasma/Serum - HPLC

    Details
  3. Precipitation Reagent
    Precipitation Reagent
    Order no.: 31004/RUO

    Vitamin B6 in Plasma/Serum - HPLC

    Details
  4. Neutralisation Reagent
    Neutralisation Reagent
    Order no.: 31005/RUO

    Vitamin B6 in Plasma/Serum - HPLC

    Details
  5. Derivatisation Reagent
    Derivatisation Reagent
    Order no.: 31006/RUO

    Vitamin B6 in Plasma/Serum - HPLC

    Details
  6. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  1. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  1. 31100 HPLC column vitamin B6 plasma serum whole blood
    HPLC column
    Order no.: 31100

    Vitamin B6 in Whole Blood/Serum/Plasma

    Details
  2. PEEK-encased Prefilter, 5 µm
    Details
  3. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  4. Precolumn Cartridge Holder 4/10
    Details
  5. Precolumn cartridge 4/10
    Precolumn cartridge 4/10
    Order no.: 18031/RUO

    Vitamin B6 in Plasma/Serum - HPLC

    Details