王曼等

摘 要 應用在線固相萃取（SPE）高效液相色譜（HPLC）方法研究TEB415在小鼠體內的藥代動力學。通過在線SPEHPLC方法結合Ultimate3000系統(tǒng)測定TEB415血藥濃度，4 結 論

本研究開發(fā)的在線固相萃取高效液相色譜方法可準確、快速測定小鼠血漿中TEB415的濃度，成功應用于TEB415的藥代動力學研究。實驗結果表明， TEB415具有藥物吸收程度較高、吸收速度適中、半衰期適中、體內消除速度適中的藥代動力學特點。本方法無需手動預處理樣品過程，避免了柱前繁瑣的分離提取過程，大大縮短了分析時間，既排除內源性物質干擾，提高方法回收率，同時又對血漿中目的成分進行了濃縮，提高了分析靈敏度。在線SPEHPLC方法具有進樣量少、分析時間短、SPE柱可多次使用、靈敏度高等優(yōu)點，可廣泛應用于生物樣品分析。

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Application of Online SPEHPLC System in Pharmacokinetic

Study of Highly Active AntiCancer Compound TEB415

WANG Man1， WEN YaBin2， LIU KangNing1， SI Ge3， LIU Lei1， YIN Zheng1， LU YaXin*1

1（School of Pharmacy， Nankai University， Tianjin 300071， China）

2（School of Life Science， Nankai University， Tianjin 300071， China）

3（School of Chemistry， Nankai University， Tianjin 300071， China）

Abstract An online solid phase extractionhigh performance liquid chromatography （SPEHPLC） system was applied in the plasma pharmacokinetic study of highly active anticancer compound tyrosine kinase inhibitors （TEB415） in mouse. The online SPEHPLC method associated with Ultimate3000 system which was applied to the determination of the blood drug level of TEB415 in mouse plasma. C18 column （Venusil MP， 150 mm × 4.6 mm， 5 μm） was used as analytical column and the mobile phase consisted of acetonitrile5 mmol/L monopotassium phosphate buffer （pH 3.5） at a flow rate of 1.0 mL/min was used as the isocratic elution. An MF Ph1 column （10 mm×4 mm， 5 μm） was used as online SPE column， and water and wateracetonitrile were used as the washing solvent and elution solvent respectively. The detection wavelength was set at 262 nm. The pharmacokinetic parameters were calculated by WinNonlin 5.2 software. The linear range of the calibration curve was between 100 and 20000 μg/L， and the limit of qualification was 20 μg/L. The extraction recovery was between 90.5% and 94.6%. The RSD of intraday and interday precision was less than 3.5%. The accuracy of shortterm stability， freezethaw stability and longterm stability were between 91.49% and 101.96%. After oral medication， the mean peak time （Tmax） of TEB415 in mice was 5.29 h， and the mean maximum concentration （Cmax） was 3403 μg/L. The area under the curve （AUC） of TEB415 was 24600 μg/L·h. This drug′s mean halflife was 3.84 h， and its mean retention time （MRT） was 6.56 h. These parameters suggested that TEB415 had appropriate rate of absorption and elimination with preferable bioavailability.

Keywords Online solid phase extraction； High performance liquid chromatography； Tyrosine kinase inhibitors TEB415； Pharmacokinetics； Blood drug level

（Received 30 August 2014； accepted 24 October 2014）

This work was supported by the Natural Science Foundation of Tianjin of China （No.12JCQNJC08500）

3（School of Chemistry， Nankai University， Tianjin 300071， China）

Abstract An online solid phase extractionhigh performance liquid chromatography （SPEHPLC） system was applied in the plasma pharmacokinetic study of highly active anticancer compound tyrosine kinase inhibitors （TEB415） in mouse. The online SPEHPLC method associated with Ultimate3000 system which was applied to the determination of the blood drug level of TEB415 in mouse plasma. C18 column （Venusil MP， 150 mm × 4.6 mm， 5 μm） was used as analytical column and the mobile phase consisted of acetonitrile5 mmol/L monopotassium phosphate buffer （pH 3.5） at a flow rate of 1.0 mL/min was used as the isocratic elution. An MF Ph1 column （10 mm×4 mm， 5 μm） was used as online SPE column， and water and wateracetonitrile were used as the washing solvent and elution solvent respectively. The detection wavelength was set at 262 nm. The pharmacokinetic parameters were calculated by WinNonlin 5.2 software. The linear range of the calibration curve was between 100 and 20000 μg/L， and the limit of qualification was 20 μg/L. The extraction recovery was between 90.5% and 94.6%. The RSD of intraday and interday precision was less than 3.5%. The accuracy of shortterm stability， freezethaw stability and longterm stability were between 91.49% and 101.96%. After oral medication， the mean peak time （Tmax） of TEB415 in mice was 5.29 h， and the mean maximum concentration （Cmax） was 3403 μg/L. The area under the curve （AUC） of TEB415 was 24600 μg/L·h. This drug′s mean halflife was 3.84 h， and its mean retention time （MRT） was 6.56 h. These parameters suggested that TEB415 had appropriate rate of absorption and elimination with preferable bioavailability.

Keywords Online solid phase extraction； High performance liquid chromatography； Tyrosine kinase inhibitors TEB415； Pharmacokinetics； Blood drug level

（Received 30 August 2014； accepted 24 October 2014）

This work was supported by the Natural Science Foundation of Tianjin of China （No.12JCQNJC08500）

3（School of Chemistry， Nankai University， Tianjin 300071， China）

Abstract An online solid phase extractionhigh performance liquid chromatography （SPEHPLC） system was applied in the plasma pharmacokinetic study of highly active anticancer compound tyrosine kinase inhibitors （TEB415） in mouse. The online SPEHPLC method associated with Ultimate3000 system which was applied to the determination of the blood drug level of TEB415 in mouse plasma. C18 column （Venusil MP， 150 mm × 4.6 mm， 5 μm） was used as analytical column and the mobile phase consisted of acetonitrile5 mmol/L monopotassium phosphate buffer （pH 3.5） at a flow rate of 1.0 mL/min was used as the isocratic elution. An MF Ph1 column （10 mm×4 mm， 5 μm） was used as online SPE column， and water and wateracetonitrile were used as the washing solvent and elution solvent respectively. The detection wavelength was set at 262 nm. The pharmacokinetic parameters were calculated by WinNonlin 5.2 software. The linear range of the calibration curve was between 100 and 20000 μg/L， and the limit of qualification was 20 μg/L. The extraction recovery was between 90.5% and 94.6%. The RSD of intraday and interday precision was less than 3.5%. The accuracy of shortterm stability， freezethaw stability and longterm stability were between 91.49% and 101.96%. After oral medication， the mean peak time （Tmax） of TEB415 in mice was 5.29 h， and the mean maximum concentration （Cmax） was 3403 μg/L. The area under the curve （AUC） of TEB415 was 24600 μg/L·h. This drug′s mean halflife was 3.84 h， and its mean retention time （MRT） was 6.56 h. These parameters suggested that TEB415 had appropriate rate of absorption and elimination with preferable bioavailability.

Keywords Online solid phase extraction； High performance liquid chromatography； Tyrosine kinase inhibitors TEB415； Pharmacokinetics； Blood drug level

（Received 30 August 2014； accepted 24 October 2014）

This work was supported by the Natural Science Foundation of Tianjin of China （No.12JCQNJC08500）