ABSTRACT
Objective: To develop a rapid, simple, sensitive, and accurate high performance liquid chromatography-tandem mass spectrometry (HPLC-MS) method for determining the contents of cinobufagin and resibufogenin from Bufo Corium. Methods: The contetns were determined by Zorbax SB-C18 column and tested in positive electrospray ionization with multiple reaction monitoring of HPLC-MS. The mobile phase was conswasted with the mixture of acetonitril-0.1 mol/L formic acid water solution (41:59), flow rate 200 μL/min, column temperature 30 oC. Mass spectrometry conditions: nitrogen temperature 350 oC, nitrogen flow rate 12 L/min, nebulizer pressure 101.316 kPa (35 psi). The shredded voltage of cinobufagin was 160 V, the collwasion energy was 15, the parent ion and the daughter ion were 443.2 and 364.8 respectively; The fragmentation voltage of resibufogenin was 130 V, the collwasion energy was 15, and the parent ion and the daughter ion were 385.2 and 366.9 respectively. Results: There was good linearity in the range of 0.99-7.92 μg/mL and 1.04-8.32 μg/mL for the cinobufagin and resibufogenin. The detection limit (S/N ≥ 3) was 0.3 ng/g. The recoveries of cinobufagin and resibufogenin ranged from 97.96% to 103.7% and 96.86% to 102.4%, respectively. The intraday and daytime precwasion were both less than 3%. Conclusion: The results showed that the method was sensitive and reliable, which can meet the needs of analyse of toxic substances in B. Corium.