Simultaneous determination of 8 bufo toxins in toad skin by UPLC/Q-TOF-MS

Abstract

ObjectiveTo establish a rapid qualitative and quantitative method for the determination of 8 bufo toxins in toad skin residual from poisoned patients by ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry （UPLC/Q-TOF-MS）.MethodsAfter the toad skin samples were pretreated， the sample solution was separated on A Cquity UPLC® HSS T3 column （100 mm×2.1 mm， 1.8 μm） with 0.1% formic acid solution as mobile phase A and methanol as mobile phase B. The flow rate was 0.3 mL/min， the column temperature was 35 ℃， and the sample volume was 2 μL.ResultsThe linear ranges were 10.0-1 000.0 μg/L with r2≥0.990. The limits of detection were 0.7-2.1 μg/kg， and the limits of quantification were 2.5-7.0 μg/kg. The average recoveries were ≥75.79% and RSDs were ≤9.84%. The contents of bufalin， desacetylcinobufagin， desacetylcinobufotalin， pseudobufarenogin and bufotaline were 123.8±3.1， 5 920.4±1.9， 680.7±3.2， 671.6±9.1 and 1 796.6±74.9 μg/kg， respectively， while gamabufotain， cinobufagin and cinobufotalin were not detected.ConclusionThe method is practical， rapid and reliable for the determination of cinobufagin， bufalin， pseudobufarenogin， desacetylcinobufotalin and desacetylcinobufagin， bufotaline， gamabufotain and cinobufotalin in toad skin.

Keywords

• ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry
• bufo toxin
• simultaneous determination