HPLC Separation of Vitamin B

    April 3, 2004

    Primesep C separates B vitamins with baseline resolution by a combination of cation exchange, complex formation, and hydrophobic interactions. Vitamin B1 (thiamine), vitamin B2 (riboflavin), and vitamin B6 (pyridoxine) are separated with a mobile phase of water, acetonitrile (MeCN, ACN) and triethylamine (TEA) phosphate with UV detection at 280 nm.


    Column Primesep C , 4.6×150 mm, 5 µm, 100A
    Mobile Phase MeCN/H2O  -10/90%
    Buffer NEt3H3PO4 Ph 3.0 – 20 Mm
    Flow Rate 1.0 ml/min
    Detection UV, 280 nm



    Class of Compounds
    Drug, Vitamin B₆, Hydrophobic, Ionizable
    Analyzing Compounds Pyridoxine, Thiamine, Riboflavin

    Application Column

    Primesep C

    The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

    Select options
    Application Analytes:

    Vitamin B1 (Thiamine)
    Vitamin B2 (Riboflavin)
    Vitamin B6 (Pyridoxine)

    Application Detection:
    UV Detection
    SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to so we can ensure you get optimal results for your compound/s of interest.