Quadruplex stable isotope derivatization strategy for the determination of panaxadiol and panaxatriol in foodstuffs and medicinal materials using ultra high performance liquid chromatography tandem mass spectrometry.

This work proposed a novel strategy based on quadruplex stable isotope derivatization for sensitive, accurate, speedy and green determination of panaxadiol (PD) and panaxatriol (PT). This strategy integrated with ultrasound-assisted dispersive liquid-liquid microextraction, microwave-assisted derivatization, and magnetic graphene oxide dispersive solid phase extraction as efficient sample pretreatment techniques, coupling with ultra-high performance liquid chromatography tandem mass spectrometry detection in multiple reaction monitoring mode for the analytical purpose of high-throughput, sensitivity, selectivity, green and accuracy. Quadruplex mass spectrometry sensitizing derivatization reagents, 3-N-(D0-/D3-methyl-, and D0-/D5-ethyl-)-2'-carboxyl chloride rhodamine 6 G, were designed, synthesized and applied for the high-throughput derivatization of hydroxyl‑containing PD and PT for the first time. The analogue derivatives of PD and PT standards using 3-N-propyl-2'-carboxyl chloride rhodamine 6 G as derivatization reagent were employed as the internal standards for the mass spectrometry quantitation to minimize matrix effect. All rhodamine 6 G derivatives of PD and PT were easily and selectively adsorbed by magnetic graphene oxide for efficient magnetic dispersive solid phase extraction before injection. Parameters of (micro-) extractions, derivatization and liquid chromatography-mass spectrometry conditions were all optimized in details. Method validation results consisting of linearity, accuracy, precision, repeatability, matrix effect, stability and limits of detection (LODs, 0.02 ng/mL for PD and 0.03 ng/mL for PT) were determined. The analytical performance of this method benefited from efficient integration of suitable sample pretreatment techniques and characteristic structure of rhodamine 6 G with four mass tags in derivatization reagents. The developed and validated method has been applied to the quantitation of PD and PT for the quality assessment of related foodstuffs and traditional Chinese medicinal materials.