Vitamin K kit

VITAMIN K IN SERUM/PLASMA LC-MS/MS ANALYSIS KIT

Vitamin K is a cofactor for enzymatic modification of glutamic acid residues (Glu) to gammacarboxyglutamic acid residues (Gla) in vitamin K-dependent Gla proteins. These Gla proteins are necessary for hemostasis, bone metabolism, vascular calcification and cell proliferation. Vitamin K is found in nature as phylloquinone (vitamin K1) in green leafy vegetables, algae and some plant oils, and menaquinones (vitamin K2 also termed MK-n, where n is the number of isoprenoid units) can be found in meat, eggs and fermented food. A synthetic form of vitamin K menadione (vitamin K3) can be found as an additive in some animal feeds.

  • Highlights

    • Simultaneous measurement the concentrations of 5 forms of vitamin K

    • Total run time is 12 min.
    • One sample preparation for all forms easy liquid-liquid extraction step following by protein precipitation for the sample treatment- no need for SPE
    • Long life span of HPLC column
  • Matrix

    • Plasma
    • Serum
  • Parameter

    Vitamin K1-Phylloquinone, Vitamin K1-2,3-Epoxide, Vitamin K2-7-Menaquinone, Vitamin K2-
    2,3-Epoxide and Vitamin K2-4-Menaquinone

Sample Preparation


  • Step 1

    Pipette 400 µL of serum sample/working calibration standard into a glass centrifuge tube

  • Step 2

    Add 800 µL of Reagent 1, additionally vortex for 5 sec.

  • Step 3

    Add 25 µL of internal standard working stock solution, then 2 ml Reagent-2 and vortex for 3 sec.

  • Step 4

    Centrifuge at 4000 rpm for 5 min.

  • Step 5

    Transfer 2.4 ml of the upper phase to a new glass tube (tube-3) and evaporate under nitrogen stream. (Upper phase can be evaporated at 7 psi during ~15 min. using water bath at the temperature of 30°C)

  • Step 6

    Reconstitute tube-3 with 100 µl of Reagent-3 and transfer to a HPLC vial prior to injection

  • ION Chromatogram

ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..

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