High-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry for the determination of flocoumafen and brodifacoum in whole blood.
A high-performance liquid chromatographic-tandem mass spectrometric (HPLC-MS-MS) assay was developed and validated to determine quantitatively flocoumafen and brodifacoum in whole blood using warfarin as an internal standard (IS). Liquid-liquid extraction, using ethyl acetate, was used to isolate flocoumafen, brodifacoum and the IS from the biological matrix. Detection was performed on a mass spectrometer by negative electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The calibration curves were linear (r(2) > 0.998) in the concentration range of 0.1-100.0 ng ml(-1) with a lower limit of quantification of 0.05 ng ml(-1) for flocoumafen, and 0.1 ng ml(-1) for brodifacoum in whole blood. Intra-day and inter-day relative standard deviations (RSDs) were less than 8.0% and 10.8%, respectively. Recoveries of flocoumafen and brodifacoum ranged from 78.0% to 83.7%. This assay can be used to determine trace flocoumafen and brodifacoum in whole blood to investigate suspected poisoning of human and animals.
Jin
Mi-Cong
MC
Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China. jmcjc@nbip.net
OuYang
Xiao-Kun
XK
Chen
Xiao-hong
XH
D015110:4-Hydroxycoumarins / Q000097:blood* / Q000737:chemistry; D000925:Anticoagulants / Q000097:blood* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D016903:Drug Monitoring / Q000379:methods; D004355:Drug Stability; D053593:Forensic Toxicology / Q000379:methods; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards; D014859:Warfarin / Q000097:blood
D015110:4-Hydroxycoumarins; D000925:Anticoagulants; C048069:flocoumafen; D014859:Warfarin; C013418:bromfenacoum