Development and Validation of a Method to Simultaneously Determine Multiple Sterols in Diversiform Food Substrates with UPLC-MS/MS

Sterols play an essential part in physiological activities of almost all creatures. However, there is a lack of a widely applicable routine method to investigate various sterols with both saturated and unsaturated forms in diversiform food materials. In this study, a new UPLC-MS/MS method with wider extensive applicability has been developed to fill this blank, which is considered time and cost saving, and can improve labor efficiency. The Agilent Eclipse Plus C 18 column and methanol-water gradient elution system were employed to improve the separation at 40 °C and the flow rate was 0.5 mL/min. The method validation parameters include precision, recovery, linearity, and limit of detection and quantification (LOD and LOQ). The recoveries of sterols in all food matrices and the inter-day and intra-day precisions represented by RSDs were all within the recommended range of chemical analysis. The LODs and LOQs in different types of samples were in range of 0.0036–0.68 μg/mL and 0.01–2.0 μg/mL, respectively. All the results demonstrated that this method was sensitive and accurate enough. Additionally, the acid hydrolysis was found closely related to the degradation of unsaturated sterols in the pretreatment.