Rapid determination of three anticoagulant rodenticides in whole blood by liquid chromatography coupled with electrospray ionization mass spectrometry.
A rapid, sensitive and selective method for the simultaneous determination of bromadiolone, flocoumafen and brodifacoum in whole blood using warfarin as internal standard (IS) by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC/ESI-MS) has been developed and validated. The target compounds were extracted from the whole blood with ethyl acetate and separated on an XDB C18 column (150 mm x 2.1 mm i.d. x 5 microm) by using a mobile phase consisting of 0.2% acetic acid/methanol (12/88, v/v) at a constant flow rate of 0.50 mL/min. The analytes were detected using negative ESI-MS in the selected ion monitoring (SIM) mode. The molecular ions [M-H]- of m/z 527, 541,523 and 307 were selected for the quantification for bromadiolone, flocoumafen, brodifacoum and the IS, respectively. The calibration curves were linear (r2 > 0.995) in the concentration range of 0.50-100.00 ng/mL. The method showed a satisfactory sensitivity (0.05-0.5 ng/mL using 200 microL blood), precision (RSD < 11.9%), accuracy (recovery: 82.0-96.1%) and selectivity. This method was successfully applied to the determination of the analytes for the diagnoses of poisoned human beings and animals.
Jin
Mi-Cong
MC
Ningbo Municipal Center for Disease Control and Prevention, 237 Yongfeng Road, Ningbo 315010, China. jmcjc@nbip.net
Chen
Xiao-Hong
XH
D015110:4-Hydroxycoumarins / Q000097:blood*; D000925:Anticoagulants / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D015203:Reproducibility of Results; D012378:Rodenticides / Q000097:blood*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D015110:4-Hydroxycoumarins; D000925:Anticoagulants; D012378:Rodenticides; C048069:flocoumafen; C013418:bromfenacoum; C021636:bromadiolone