Investigation of Retention Mechanism of B2 and B3 in Home-Made ZIC-HILIC Columns and Its Application for The Simultaneous Estimation in Multivitamin Tablets

Abstract

In the present study, the retention behavior of two vitamins (B₂ and B₃) was investigated using a zwitterionic hydrophilic interaction liquid chromatography technique (ZIC-HILIC) on two home-made columns, and two analytical methods for these vitamins were established as a result. Several chromatographic parameters were carefully investigated and optimized. The best separation was achieved on two new home-made columns (ZIC-1 and ZIC-3) employing acetonitrile/sodium acetate buffer as the mobile phase (40 mM, pH 4.75). The target vitamins were detected at 254 nm. In the ZIC-1 and ZIC-3 columns, the linear ranges of the target vitamins (B₂ and B₃) were 0.06-11.0 and 0.08-7.0 µg/mL, respectively. For both target vitamins, the detection and quantitation limits of the suggested methods were 0.011-0.055 and 0.033-0.166 µg/mL, respectively. They also showed high precision (RSD ≤ 0.93), linearity (r² ≥ 0.9988), and recovery (between 99.0 and 101.71%). The suggested procedures have been validated and tested on multivitamin tablets, proving that they are suitable for analyzing food supplements for these tablets, and acceptable results were obtained for the study.