Simple and Sensitive Liquid Chromatography–tandem mass Spectrometry Methods for Quantification of Paraquat in Plasma and Urine: application to experimental and clinical toxicological studies

Abstract

Simple, sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) methods have been developed and validated for quantification of paraquat (PQ) in plasma and urine. Plasma and urine sample preparation were carried out by one-step protein precipitation using cold acetonitrile (−20 to −10 °C). After centrifugation, an aliquot of 10 μL of supernatant was injected into a Kinetex™ hydrophilic interaction chromatography (HILIC) column with a KrudKatcher™ Ultra in-line filter. The chromatographic separation was achieved using the mobile phase mixture of 250 mM ammonium formate (with 0.8% aqueous formic acid) in water and acetonitrile at a flow rate of 0.3 mL/min. Detection was performed using an API2000 triple quadrupole tandem mass spectrometer in multiple reaction monitoring (MRM) mode via an electrospray ionization (ESI) source. The calibration curve was linear over the concentration range of 10–5000 ng/mL, with an LLOQ of 10 ng/mL. The inter- and intra-day precision (% R.S.D.) were <8.5% and 6.4% for plasma and urine, respectively with the accuracies (%) within the range of 95.1–102.8%. PQ in plasma and urine samples was stable when stored at −70 °C for three freeze–thaw cycles. The methods were successfully applied to determine PQ concentration in rat and human samples.