Application of direct protein precipitation combined with tandem mass spectrometry in monitoring 5-fluorouracil therapeutic drugs in plasma
To develop a method for the determination of 5-fluorouracil in plasma by direct protein precipitation coupled with tandem mass spectrometry.Plasma samples were treated with 0.1 mol·L-1 zinc sulfate as protein precipitator,and the supernatant was taken after centrifugation for direct determination.Using 5-fluorouracil-13C,15N2 as internal standards and Shimadzu shim pack GIST-HP C18-AQ(100 mm × 2.1 mm I.D.,1.9 μm)as chromatographic column;Water(phase A)-methanol(phase B)as mobile phase;Gradient elution:0-1 min,5％B;1-2 min,5％B→95％B;2-2.5 min,95％B;2.5-2.6 min,95％B→5％B;2.6-5 min,5％B;Flow rate:0.3 mL·min-1;Column temperature:35 ℃;Injection volume:5 μL;Mass spectrometry quantitative analysis was carried out by using ESI and multi reaction monitoring negative ion mode(MRM);Detection ion pair:5-fluorouracil m/z 129.1→42.1;5-fluorouracil-13C,15N2 internal standard m/z 132.1→44.1.The correlation coefficient was greater than 0.999.The determination accuracy of the quality control was close to the theoretical v(el)ue.The intra batch and inter batch accuracy were in the range of 94.8％—99.0％,and the RSD were in the range of 0.9％-4.8％.The recovery of 5-fluorouracil were in the range of 95.6％—99.8％.The matrix factor were in the range of 0.92-0.96,and the normalized matrix factor of internal standard were in the range of 0.99-1.00.The relative deviation RE between each measured concentration and the marked concentration under all conditions of the stability test were in the range of-5.2％—1.7％.This method has the advantages of simple pretreatment,fast analysis speed,high sensitivity,specificity and good stability,and can be used for the monitoring 5-fluorouracil.
direct protein precipitationtandem mass spectrometry5-fluorouraciltherapeutic drug monitoring
NETL
NSTL
万方数据
