Determination of Amanita Peptides in Human Plasma and Urine by High-Performance Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry

Five amanita peptide toxins, such as alpha-amanita, beta-amanita, phalloidin, viroidin and ala-viroidin in human plasma and urine were determined by high-performance liquid chromatography coupled with triple quadrupole mass spectrometry under positive ion multiple reaction monitoring mode (MRM). The samples were enriched and purified by HLB solid phase extraction cartridge, and separated by CAPCELL CORE C-18 column using 10 mmol/L aqueous ammonium acetate-methanol as mobile phase under a gradient elution. The results indicated that the five amanita peptide toxins in plasma or urine had good linear relationship in a wide range of concentration levels. The detection limits of the toxins were 0.2 and 0.5 mu g/L (S/N = 3), the average recoveries were 98.0% - 114.6% and 90.1% - 99.4% for spiked plasma and urine samples, respectively. Accuracy, precision and matrix effects could meet the methodological requirement for biological samples analysis. The established method had many advantages such as simple operation and high sensitivity, which provided an accurate and reliable technical support for the diagnosis and medical treatment of clinical amanita poisoned patients.