Objective: To establish an analytical method based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of imperialine-3β-D-glucoside

imperialine

peimisine

verticine

verticinone and hupehenine in Fritillaria unibracteata var. wabuensis. Method: The separation was carried out on Agilent Eclipse XDB C18 (2.1 mm×100 mm

1.8 μm) with 0.1% formic acid solution (containing 0.01 mol·L-1ammonium formate)-acetonitrile as mobile phase for gradient elution at a flow rate of 0.4 mL·min-1

and column temperature was 35℃. The target components were detected in multiple reaction monitoring mode by mass spectrometry with electrospray ionization (ESI) source under positive mode. The quantitative and qualitative ion pairs of the six analytes were as follows:imperialine-3β-D-glucoside m/z 592.4 > 574.4

138.2

96.1

imperialine m/z 430.4>412.3

138.2

96.1

peimisine m/z 430.4>412.3

109.2

67.2

verticine m/z 432.4>414.4

95.2

67.2

verticinone m/z 430.4>412.3

396.3

91.1

and hupehenine m/z 416.4>98.2

81.2

67.2.The results showed that good linearity was achieved in their respective linear ranges (r>0.999 1)

with good precision

repeatability and stability. The average recovery ranged from 98.07% to 106.62%

and the RSD was less than 5%. Result: There were great differences in the content of 6 components between various batches. In general

the contents were highest for imperialine and imperialine-3β-D-glucoside

followed by peimisine

verticinone

verticine and hupehenine progressively. Conclusion: The method is sensitive and accurate

and suitable for the simultaneous determination of multiple components in Fritillaria unibracteata var. wabuensis

providing a basis for its quality control and comprehensive assessment.