Development of a simple and reliable method for α-amanitin detection in rat plasma and its application to a toxicokinetic study.
Rapid Commun Mass Spectrom
; 35(21): e9184, 2021 Nov 15.
Article
in En
| MEDLINE
| ID: mdl-34472672
ABSTRACT
RATIONALE α-Amanitin is a highly toxic peptide widely found in species of poisonous mushrooms. The matrix effect has been a major obstacle for accurate determination of α-amanitin in plasma samples by liquid chromatography/tandem mass spectrometry (LC/MS/MS). In this study, the strategy to eliminate the matrix effect of α-amanitin with a one-step dilution approach after deproteinization was applied. METHODS:
Rat plasma samples were processed by protein precipitation with methanol followed by a nine-fold dilution with pure water. The matrix effect value of α-amanitin was 19.7%-22.2% by protein precipitation and then changed to 87.5%-88.7% after dilution. α-Amanitin and the internal standard (roxithromycin) were analyzed on an ACQUITY UPLC® BEH C18 (50 mm × 2.1 mm, 1.7 µm) column within 3.0 min by gradient elution.RESULTS:
The linear ranges were 0.90-600 ng/mL with a correlation coefficient r >0.9958. A lower limit of quantification (LLOQ) of 0.90 ng/mL was achieved using only 50 µL of rat plasma. The intra- and inter-day precisions for the analyte ranged from 3.2% to 7.5% and 3.1% to 7.1%, respectively, and the accuracy ranged from -5.3% to -8.0%.CONCLUSIONS:
The matrix effect of α-amanitin was reduced by sample dilution after plasma deproteinization. A reliable LC/MS/MS method for the determination of α-amanitin in rat plasma was developed. This method was successfully applied for a toxicokinetic study of rats after intravenous injection of α-amanitin with a subacute toxicity dose at 0.10 mg/kg.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Chromatography, Liquid
/
Tandem Mass Spectrometry
/
Alpha-Amanitin
Type of study:
Diagnostic_studies
/
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Rapid Commun Mass Spectrom
Year:
2021
Document type:
Article
Affiliation country:
China