Development and application of a strategy for analyzing eight biomarkers in human urine to verify toxic mushroom or ricinus communis ingestions by means of hydrophilic interaction LC coupled to HRMS/MS

Further development of a liquid chromatography–high‐resolution mass spectrometry/mass spectrometry‐based strategy for analyzing eight biomarkers in human urine indicating toxic mushroom or Ricinus communis ingestions

“…29,[36][37][38][39][40][41][42][43][44] Tomkov a et al developed a method for a simultaneous determination of αand β-amanitin and muscarine in urine by LC-high-resolution (HR)-time-of-flight (TOF) MS. 45 Recently, we presented a comprehensive and validated analytical solution to identify a total of eight biomarkers of castor bean intoxication, phalloides syndrome, pantherine-muscaria syndrome, muscarine syndrome, psilocybin syndrome, and bufotenine intoxication in urine within one single run. 18 The samples were prepared by SPE and urine precipitation performed in parallel and the use of a four-eluent system for normal-phase chromatography were necessary to realize the aims of selectivity and earmarked sensitivity for all analytes. However, as there is not always the need to screen for all biomarkers using such an extensive method, the method should be simplified by saving cost and turnaround time while enhancing flexibility in toxicological analysis by development of two separate methods for biomarker identification of late-and early-onset syndromes, respectively.…”

“…Ricinine showed a much higher recovery (106%) than in Method R (33%), whereas recoveries of muscarine and muscimol appeared to be lower than in the referenced method (110% and 68%, respectively). 18 The latter could be the result of muscarine and muscimol recovery in the SPE fraction of Method R, not existing in Method B.…”

“…The instrument was the same as for Method R. 18 Eluent B was 100% ACN and eluent C purified water. The multistep gradient consisted of six time windows (TWs) as follows: TW 1, step: from 0:00 to 0:20 min, A: kept at 4%, B: kept at 96%; TW 2, ramp: from 0:20 to 8:20 min, A: to 12%, B: to 88%; TW 3, ramp: from 8:20 to 9:20 min, A: to 28%, B: to 72%; TW 4, ramp: from 9:20 to 12:50 min, A: to 40%, B: to 60%, C: to 0%; TW 5, step: from 12:50 to 15:20 min, A: kept at 25%, B: kept at 25%, C: kept at 50%; TW 6, step: from 15:20 min to 16:50 min, A: kept at 4%; B: kept at 96%, C kept at 0%; the flow rate was set to 0.80 ml/min in TW 1, 0.75 ml/min in TW 2, 0.70 ml/min in TW 3, 0.60 ml/min in TW 4 and 5, and 0.50 ml/min in TW 6.…”

Analysis of α- and β-amanitin in Human Plasma at Subnanogram per Milliliter Levels by Reversed Phase Ultra-High Performance Liquid Chromatography Coupled to Orbitrap Mass Spectrometry

“…Before SPE was performed, the A-LLE extract had to be acidified because β-amanitin was only retained in the SPE column when in the protonated (uncharged) form. Strata X-Drug B cartridges (mixed-mode strong cation-exchange resin) and a similar SPE protocol had been used successfully in a previous study [30]. In the current study, we determined an extraction recovery of 50.2% (coefficient of variation (CV) = 11.4%), 55.7% (CV = 8.4%), and 87.6% (CV = 19.8%) for α-amanitin, β-amanitin, and γ-amanitin methyl ether (internal standard), respectively.…”

“…Urine is usually the matrix of choice for such analysis due to higher concentrations compared to other body fluids [ 26 ]. Several methods were published for their analysis in urine by using LC-MS, LC-(HR)MS/MS, capillary zone electrophoresis, HPLC, radioimmunoassay, ELISA, and recently a lateral flow immunoassay [ 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 ]. Also, blood plasma or serum can be used as demonstrated earlier [ 33 , 34 , 38 , 39 , 40 , 41 ].…”