Simultaneous determination of 14 biogenic amines and metabolites in bullfrog blood using UFLC-MS/MS with SPE

Abstract

In this work, a reliable, accurate, and sensitive method based on a simple extraction with trichloroacetic acid at 5% and a clean-up step by means of a Strata-X SPE cartridge prior to the analysis of 14 biogenic amines (BAs) and metabolites by ultra-fast liquid chromatography-tandem quadrupole mass spectrometry (UFLC-MS/MS) was developed. The 14 BAs and metabolites could be separated by using a LC column under conditions of gradient elution within 6.0 min and simultaneously determined without interference from contaminants in biological samples. The results showed that the Strata-X SPE cartridge used in this method is more effective than C18 SPE and HLB SPE cartridges for cleanup of BAs and metabolites in bullfrog blood. The obtained results demonstrated the higher extraction capacity of the Strata-X SPE cartridge with recoveries between 73.2 and 102%. The limits of quantification for the 14 BAs and metabolites ranged from 0.20–34.3 μg L⁻¹. The developed Strata-X cartridge SPE UFLC-MS/MS method had been successfully applied to 15 real samples, and it was confirmed that the Strata-X SPE cartridge was highly effective for clean-up of BAs and metabolites.