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Vitamin D in Serum/Plasma HPLC Analysis Kit

Vitamin D plays a vital role in skeletal metabolism, calcium homeostasis, and also in the functioning of the immune, cardiovascular, and reproductive systems. Vitamin D deficiency leads to rickets and osteomalacia and is also associated with breast and colorectal cancers, multiple sclerosis, dementia, rheumatoid arthritis, diabetes, parkinson’s and alzheimer’s diseases.

Consequently, laboratory testing of quantitative HPLC analysis kit for 25-Hydroxyvitamin D2 and D3 in human serum & plasma samples has experienced phenomenal growth over the past several years.

  • Highlights

    Separation of 25-Hydroxyvitamin D2 and D3.

    Simple sample preparation without SPE or evaporation.

    5.0 minutes analysis time.

    Isocratic mobile phase gradient.

    Long lifetime HPLC column
  • Parameters

    25-Hydroxyvitamin D2 & D3
  • Matrix

    Serum, Plasma

Sample Preparation

  • Step 1
    Take 475 μl serum & plasma sample into a centrifuge tube
  • Step 2
    Add 500 μl of Reagent-1 and vortex for 5 seconds
  • Step 3
    Add 500 μl of Reagent-2 and vortex for 15 seconds. Then, centrifuge at 10000-12000 rpm for 5 minutes.
  • Step 4
    Decant the clear supernatant into HPLC vial.
  • Chromatogram


    Analytes LOQ Accuracy Repeatability Linearity
    (µg/L) (%) (%RSD) (R2)
    25-Hydroxyvitamin D2 5,53 90,8 3,16 0,9993
    25-Hydroxyvitamin D2 4,17 88,7 3,44 0,9991

    All results were obtained using Agilent DAD dedector and 1200 series HPLC system in SEM R&D lab.

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