Quantitative determination of coumatetralyl in whole blood with UPLC-QTRAP-MS/MS

Objective To establish a simply, sensitive and accurate quantitative method for determination of coumatetralyl in blood samples.

Methods The quantitative method of UPLC-QTRAP-MS/MS for the determination of coumatetralyl was developed. Coumatetralyl was extracted from whole blood samples by liquid-liquid extraction with ethyl acetate. The samples were extracted with ethyl acetate, then gradient eluted with ammonium acetate (5 mmol/L) -methanol as mobile phase and separated on C₁₈ column. Electrospray ionization, multiple reaction monitoring and enhanced product ion scan mode were used for mass spectrometry detection.

Results The detection limit of the method was 3.4 ng/mL. The working curve had a good linearity in the range of 10-1 200 ng/mL. The average recoveries of the three samples with different concentrations was 92.5%, 94.3% and 101.0%, respectively. Coumatetralyl in blood could be stored for at least 7 days at 4℃.

Conclusion The established quantitative method could reduce the interference of impurities, and had the advantages of simplicity, sensitivity and accuracy. It could meet the requirements of quantitative detection of coumatetralyl in blood.