Analysis Of Aristolochic Acid I In Mouse Serum And Tissues By Using Magnetic Solid-Phase Extraction And Uhplc-Ms/Ms

A method combining magnetic solid-phase extraction (MSPE) and ultra-high-pressure liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed for the analysis of aristolochic acids I (AAI) in mouse serum and tissues. The magnetic covalent organic frameworks (MNP@COF)-based MSPE exhibited high adsorption capacity towards AAI (93.1 mg/g) in optimal conditions. After MSPE extraction, AAI was separated with C18 column using gradient elution and quantified (m/z 342.21 -> 298.13) by UHPLC-MS/MS with monitor reaction monitoring (MRM) mode. This MSPE-based UHPLC-MS/MS method was validated with respected to lower limit of quantification (LLOQ), linearity, recovery, precision and accuracy of intra- and inter-day, and matrix effect. Good calibration linearities at the range of 1-500 ng/L for AAI in biological matrices (serum, kidney, and liver) with high correlation coefficient (R-2) > 0.9970, and high enrichment factors (mean values from 1038 to 1045) were obtained. This method was highly sensitive to determine AAI with LLOQ within the range of 4.62-5.24 ng/L in extracted serum, kidney, and liver samples. Recoveries at 5, 50, 100 and 300 ng/L in biological samples ranged from 93.2 to 104.0%, and intra- and inter day accuracy and precision (defined as bias and coefficient of variation, respectively) were below +/- 15%. The method was successfully applied in the analysis of biological samples collected from mice exposed with AAI with concentrations range of 0.007-0.041 mu g/L for consecutive four days. The established method might be applied for the investigation of risk assessment and toxicity induced by long-time use of AAI-containing herbs or dietary supplements.