Abstract
An innovative 4D targeted method was developed for the determination of 61 bioactive compounds in royal jelly (RJ) related to their health-promoting properties. The method, apart from high-resolution mass spectrometry, exploits the advantages of vacuum-insulated probe-heated electrospray ionization source (VIP-HESI), reducing thermal degradation, and trapped ion mobility spectrometry (TIMS), improving selectivity and compound identification. The optimization of VIP-HESI ionization parameters using experimental designs showed that the critical parameters were the capillary voltage as well as the probe gas flow rate and temperature. The LODs of the validated method ranged between 0.0013 and 0.090 μg g(-1), the recoveries between 80 and 109%, while %RSDs were below 10%. The method was applied to 22 RJ samples, and 24 bioactive compounds were determined at concentrations between 0.098 μg g(-1) (2-cis,4-trans-abscisic acid) and 18 μg g(-1) (quinic acid), while eight compounds were identified for the first time. This method serves as a reliable analytical tool for a wide-scope chemical characterization of RJ.