HPLC Method for Separation of Pyridoxine and Doxylamine on Primesep S Column


Pyridoxine, also known as Vitamin B6, is an essential nutrient required by the body to produce red blood cells and for proper nerve functioning. Doxylamine is a common treatment for insomnia. When taken together, they can treat morning sickness in pregnant women.

Pyridoxine and Doxylamine can be retained, separated, and detected on a normal-phase Primsep S using an isocratic analytical method with a simple mobile phase of water, Acetonitrile, and an Ammonium Formate buffer. This analysis method can be detected in the low UV regime at 270 nm.


Column Primesep S, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer AmFm pH 3.0- 100 mM
Flow Rate 1.0 ml/min
Detection UV 270 nm,  MS- compatible mobile phase


Class of Compounds Hydrophobic, Drug
Analyzing Compounds Pyridoxine,  Doxylamine

Application Column

Primesep S

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 B6 (Pyridoxine)
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.