Rapid HILIC method with fluorescence detection using derivatization reaction utilizing o-phthaldialdehyde for determination of degradation product of aliskiren

A rapid procedure based on direct extraction and HILIC separation of aliskiren (ALI) degradation product – 3-amino-2,2-dimethylpropanamide (ADPA) with fluorescence detection has been developed. The formation of ADPA from ALI under different conditions was studied. The evaluation of HILIC method robustness was performed using multifactorial experiments with fixed factors (one-level Plackett–Burman design). XBridge HILIC column with isocratic elution using mobile phase 10 mM K2HPO4 pH 7.2–acetonitrile (26:74; v/v) was employed. Fluorescence detection after post column derivatization using o-phthaldialdehyde (OPA) reagent was performed at excitation and emission wavelength of 345 nm and 450 nm, respectively. The reported method has an advantage of a simple sample pre-treatment and quick and very sensitive measurement. The method was successfully applied for the analysis of commercially available ALI samples.

► The formation and determination of 3-amino-2,2-dimethylpropanamide was described. ► HILIC method was optimized in order to selectively detect and quantify. ► Post-column derivatization using OPA was used for fluorescence detection.