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Simultaneous determination of concentrations of imatinib and N-desmethyl imatinib in human plasma by HPLC

LUO Xing-xian;HUANG Lin;LI Tai-feng;XUE Xue-cai;WANG Yu-hang;CHEN Yue;YANG Chang-qing;FENG Wan-yu;Department of Pharmacy,People Hospital of Peking University;School of Basic Medicine and Clinical Pharmacy,China Pharmaceutical University;  
Objective: To establish a method for determining the concentrations of imatinib and its metabolites N-desmethyl imatinib in plasma by HPLC. Methods: Blood samples were pretreated by 1 mol·L~(-1) NaOH using ethyl acetate n-hexane( 75∶ 25) with dasatinib as an internal standard,then concentrated in vacuum,and the reconstituted solution was detected by HPLC. The analytical column was ZORBAX SB-C_(18)( 250 mm ×4. 6 mm,5 μm),mobile phase was acetonitrile-potassium dihydrogen phosphate buffer( 0. 025 mol·L~(-1) KH_2PO_3,pH value was adjusted to about 3. 0 by H_2PO_3)( 27∶ 73,V/V) at a flow rate of 1. 0 mL·min~(-1). The detection wavelength was set at 267 nm,the temperature was kept at 35 ℃,and the injection volume was 20 μL. Results:The calibration curves had good linearity in the rang of 0. 10 ~ 5. 00 μg·mL~(-1) for imatinib and 0. 010 ~ 0. 05μg·mL~(-1) for N-desmethyl imatinib( r = 0. 998 and 0. 997,respectively). The limits of quantification of imatinib and N-desmethyl imatinib were 0. 1 μg·mL~(-1) and 0. 01 μg·mL~(-1),respectively. The extraction recoveries of low,medium and high concentration control samples were 86. 02%,87. 31%,88. 21% for imatinib and 81. 64%,84. 78%,88. 74% for N-desmethyl imatinib,respectively. The intra-day and inter-day RSD values were less than 15%. Conclusion: The method is simple,rapid and sensitive for drug monitoring and pharmacokinetic study on imatinib and N-desmethyl imatinib.
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