A PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure was developed for the extraction and purification of silychristins A and B, silybins A and B, isosilybins A and B, and silydianin in Silybum marianum. After optimizing the extracting solvent types and the sample loading volume, the crude extract was diluted to 3 mL with 95% acetonitrile and then loaded on the PRiME cartridge. The eluate was analyzed by ultra-performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS). All the target analytes were deprotonated as [M-H]- at m/z 481 by conducting collision-induced dissociation (CID), and the major fragment ions were m/z 463 ([M-H2O-H]-), 453 ([M-CO-H]-), 355 ([M-C6H6O3-H]-), 301 ([M355-CO2-H]-), and 179 ([C10H11O3]-). Afterwards, this method was validated in terms of linearity (R2 > 0.9990), intra-day precision (1.02%-3.79%), inter-day precision (1.59%-4.87%), sensitivity (LOD ≤ 0.45 μg·kg-1 and LOQ ≤ 1.50 μg·kg-1), and recovery (76.9-103.4%, RSD < 8.90%). Finally, the proposed protocol was successfully applied to eight batches of S. marianum samples. The total content of the seven active compounds varied amongst the batches from different places of origin.
Keywords: Lignans; PRiME; Pass-through solid-phase extraction; Silybum marianum; UPLC−MS/MS.
Copyright © 2018 Elsevier B.V. All rights reserved.