Analysis of alkaloids from Peganum harmala L. sequential extracts by liquid chromatography coupled to ion mobility...

An orthogonal two dimensional analysis method based on high performance liquid chromatography (HPLC) separation and electrospray ionization-ion mobility spectrometry (ESI-IMS) detection was developed for the analysis of alkaloid compounds from Peganum harmala L. seeds. Reverse phase (RP) and hydrophilic interaction chromatography (HILIC) were compared for the most optimal performance using three different chromatographic columns. The experimental results suggest that HILIC mode is a better option for combining with the ESI-IMS system for higher sensitivity and ease in hyphenating. Under optimized conditions, alkaloids from different extraction phases were determined by means of the established HPLC-IMS method. More compounds from Peganum harmala L. seed extracts were differentiated on the HPLC-ESI-IMS system by their retention time and drift time than by HPLC or ESI-IMS alone, and thirteen alkaloids were tentatively identified based on m/z and fragment ions using ultra-high-performance liquid chromatography tandem mass-spectrometry (UPLC-MS/MS). Hence, our results indicate that this method can be considered to be advantageous over traditional absorbance detection methods for resolving complex mixtures because of complementary separation steps, elevated peak capacity, and higher sensitivity.