在线固相萃取—高效液相色谱系统在高抗癌活性化合物TEB—415药代动力学中的应用
王曼等
摘 要 应用在线固相萃取（SPE）高效液相色谱（HPLC）方法研究TEB415在小鼠体内的药代动力学。通过在线SPEHPLC方法结合Ultimate3000系统测定TEB415血药浓度，4 结 论
本研究开发的在线固相萃取高效液相色谱方法可准确、快速测定小鼠血浆中TEB415的浓度，成功应用于TEB415的药代动力学研究。实验结果表明， TEB415具有药物吸收程度较高、吸收速度适中、半衰期适中、体内消除速度适中的药代动力学特点。本方法无需手动预处理样品过程，避免了柱前繁琐的分离提取过程，大大缩短了分析时间，既排除内源性物质干扰，提高方法回收率，同时又对血浆中目的成分进行了浓缩，提高了分析灵敏度。在线SPEHPLC方法具有进样量少、分析时间短、SPE柱可多次使用、灵敏度高等优点，可广泛应用于生物样品分析。
References
1 WANG ShaoBi. Tumor Journal of the World， 2011， 10（1）： 9-16T
王少毕. 世界肿瘤杂志， 2011， 10（1）： 9-16T
2 LIU Na， GUO DongMei， JU XueHai， CUI Xi. Chinese Journal of Pharmaceutical Analysis， 2008， 28（4）： 511-515
刘 娜， 郭冬梅， 局学海， 崔 晞. 药物分析杂志， 2008， 28（4）： 511-515
3 Li X Q， Li Y， Chen Y S. CN. Patent， WO 2014/108021 A1， 2014
4 HE Bing， TIAN Ji， LI ChunHong， AI HongBing. Chinese Journal of Pharmaceutical Analysis， 2008， 28（8）： 1316-1318
何 兵， 田 吉， 李春红， 艾红兵. 药物分析杂志， 2008， 28（8）： 1316-1318
5 CHEN Lei， ZHU JiHong， LI YongQing， LIU Wening. Chinese Journal of Pharmaceutical Analysis， 2004， 24（2）： 137-139
陈 蕾， 朱霁虹， 李永庆， 刘文英. 药物分析杂志， 2004， 24（2）： 137-139
6 Ivano M， Véronique V， Flavia B， Marc F， Martial S， Serge R， Jean L V. J. Chromatogr. A， 2010， 1217（25）： 4071- 4078
7 Ye G， Li Y Z， Li Y Y， Guo H Z， Guo D A. J. Pharm. Biomed. Anal.， 2003， 33： 521-527
8 Sheng Y X， Li L， Wang C S， Li Y Y， Guo D. J. Chromatogr. B， 2004， 806： 127-132
9 Nageswara R， Mastan V R， Dhananjay D S. Biomed. Chromatogr.， 2009， 23： 1145-1150
10 Chen L G， Yu A M， Zhuang X D， Zhang K， Wang X P， Ding L， Zhang H Q. Talanta， 2007， 74： 146-152
11 Ugo C， Fabio G， Paolo D， Eleonora M， Davide Z， Elisa R， Maria C G， Emilio M. Anal. Chem.， 2010， 82（13）： 5636-5645
12 XIE YunFeng， WANG Hao， LIU Tong， REN DanDan， YANG YongTan. Chinese J. Anal. Chem. ， 2014， 42（9）： 1343-1347
谢云峰， 王 浩， 刘 佟， 任丹丹， 杨永坛. 分析化学， 2014， 42（9）： 1343-1347
13 GUO Jian， YANG XinLei， YE MingLi. Chinese J. Anal. Chem. ， 2011， 39（8）： 1256-1260
郭 坚， 杨新磊， 叶明立. 分析化学， 2011， 39（8）： 1256-1260
14 Richard J M， Rachelle C， Heather H， Asadh M， Donald K W. J. Pharm. Biomed. Anal.， 2010， 52（1）： 86-92
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）
摘 要 应用在线固相萃取（SPE）高效液相色谱（HPLC）方法研究TEB415在小鼠体内的药代动力学。通过在线SPEHPLC方法结合Ultimate3000系统测定TEB415血药浓度，4 结 论
本研究开发的在线固相萃取高效液相色谱方法可准确、快速测定小鼠血浆中TEB415的浓度，成功应用于TEB415的药代动力学研究。实验结果表明， TEB415具有药物吸收程度较高、吸收速度适中、半衰期适中、体内消除速度适中的药代动力学特点。本方法无需手动预处理样品过程，避免了柱前繁琐的分离提取过程，大大缩短了分析时间，既排除内源性物质干扰，提高方法回收率，同时又对血浆中目的成分进行了浓缩，提高了分析灵敏度。在线SPEHPLC方法具有进样量少、分析时间短、SPE柱可多次使用、灵敏度高等优点，可广泛应用于生物样品分析。
References
1 WANG ShaoBi. Tumor Journal of the World， 2011， 10（1）： 9-16T
王少毕. 世界肿瘤杂志， 2011， 10（1）： 9-16T
2 LIU Na， GUO DongMei， JU XueHai， CUI Xi. Chinese Journal of Pharmaceutical Analysis， 2008， 28（4）： 511-515
刘 娜， 郭冬梅， 局学海， 崔 晞. 药物分析杂志， 2008， 28（4）： 511-515
3 Li X Q， Li Y， Chen Y S. CN. Patent， WO 2014/108021 A1， 2014
4 HE Bing， TIAN Ji， LI ChunHong， AI HongBing. Chinese Journal of Pharmaceutical Analysis， 2008， 28（8）： 1316-1318
何 兵， 田 吉， 李春红， 艾红兵. 药物分析杂志， 2008， 28（8）： 1316-1318
5 CHEN Lei， ZHU JiHong， LI YongQing， LIU Wening. Chinese Journal of Pharmaceutical Analysis， 2004， 24（2）： 137-139
陈 蕾， 朱霁虹， 李永庆， 刘文英. 药物分析杂志， 2004， 24（2）： 137-139
6 Ivano M， Véronique V， Flavia B， Marc F， Martial S， Serge R， Jean L V. J. Chromatogr. A， 2010， 1217（25）： 4071- 4078
7 Ye G， Li Y Z， Li Y Y， Guo H Z， Guo D A. J. Pharm. Biomed. Anal.， 2003， 33： 521-527
8 Sheng Y X， Li L， Wang C S， Li Y Y， Guo D. J. Chromatogr. B， 2004， 806： 127-132
9 Nageswara R， Mastan V R， Dhananjay D S. Biomed. Chromatogr.， 2009， 23： 1145-1150
10 Chen L G， Yu A M， Zhuang X D， Zhang K， Wang X P， Ding L， Zhang H Q. Talanta， 2007， 74： 146-152
11 Ugo C， Fabio G， Paolo D， Eleonora M， Davide Z， Elisa R， Maria C G， Emilio M. Anal. Chem.， 2010， 82（13）： 5636-5645
12 XIE YunFeng， WANG Hao， LIU Tong， REN DanDan， YANG YongTan. Chinese J. Anal. Chem. ， 2014， 42（9）： 1343-1347
谢云峰， 王 浩， 刘 佟， 任丹丹， 杨永坛. 分析化学， 2014， 42（9）： 1343-1347
13 GUO Jian， YANG XinLei， YE MingLi. Chinese J. Anal. Chem. ， 2011， 39（8）： 1256-1260
郭 坚， 杨新磊， 叶明立. 分析化学， 2011， 39（8）： 1256-1260
14 Richard J M， Rachelle C， Heather H， Asadh M， Donald K W. J. Pharm. Biomed. Anal.， 2010， 52（1）： 86-92
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）
