Separation and Retention of Vitamins in Mixed-Mode HPLC

Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Both columns allow peak order reversal by simply changing the acetonitrile concentration in the mobile phase. Vitamin B2 is especially sensitive to acetonitrile concentration. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either trifluoroacetic acid (TFA) or acetic acid (HOAc) with UV detection at 250 nm.

Condition

Column Primesep B, Primesep B2 , 3.2×50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer Formic Acid, Acetic Acid
Flow Rate 0.5 ml/min
Detection UV, 250 nm

 

Description

Class of Compounds
Drug, Vitamin B₆, Hydrophobic, Ionizable
Analyzing Compounds Vitamins C (ascorbic acid) and B2 (riboflavin)

Application Column

Primesep B

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.

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Primesep B2

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.

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Application Analytes:
Vitamin B2 (Riboflavin)
Vitamin C

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 research@sielc.com so we can ensure you get optimal results for your compound/s of interest.