Rapid Determination of Anisatin in Biological Sample by Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry

A rapid UPLC-MS/MS method was developed for the quantitative determination of anisatin,as a marker of the sesquiterpene lactones in various Illicium species,in plasma,urine,vomit,and plant samples using a triple quadrupolep mass spectrometer.The proposed assay includes multi-function impurity adsorption SPE cleanup procedure for plasma sample,diatomite solid supported liquid/liquid extraction for urine,and tert-butyl methyl ether liquid/liquid extraction for plant sample.The satisfactory recoveries were obtained.Chromatographic separation was performed on an UPLC BEH C18 column(50 mm μ 2.1 mm × 1.7 μ using gradient elution with water-methanol,and detected by the negative electrospray ionization-MS/MS method under multiple reaction monitor mode.The cycle time of each analysis was 5.0 min.The average recoveries were 92.6%-100.3% and 101%-118% with RSDs of 3.8%-11% and 6.4%-17%(n=6) for anisatin in plasma and urine,respectively.The limits of quantitation(S/N=10) were 2 μg/L and 1 μg/L for plasma and urine sample,respectively.The method is simple,selective and sensitive to the determination of anisatin in biological sample for toxicological purposes.