Quantitative determination of acetylshikonin in macaque monkey blood by LC-ESI-MS/MS after precolumn derivatization with 2-mercaptoethanol and its application in pharmacokinetic study  

作　　者：Dong-xiao SUN Hui-fang TIAN Zhi-yun MENG Alicia DU Dan YUAN Ruo-lan GU Zhuo-na WU Gui-fang DOU 

机构地区：[1]Laboratory of Drug Metabolism and Pharmacokinetics, Beijing Institute of Transfusion Medicine, Beijing 100850, China [2]School of Chinese Traditional Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China [3]ThermoFisher Scientific, San Jose, California, USA

出　　处：《Acta Pharmacologica Sinica》2008年第12期1499-1506,共8页中国药理学报（英文版）

摘　　要：Aim： To develop and validate a novel precolumn derivatization method for the quantitative determination and pharmacokinetic application of acetylshikonin in macaque monkeys by liquid chromatography-electrospray ionization tandem mass spectrometry （LC-ESI-MS/MS）. Methods： 2-Mercaptoethanol was added to the blood sample as the derivatization reagent. The derivatization reaction formed 1-major derivation product, Which was well correlated with acetylshikonin. The acetylshikonin concentrations in the biological samples were calculated by quantitative determination of the major derivation product using LCESI-MS/MS. Separation was achieved using a C18 column （2 mm×50 mm, 5μm） at room temperature and a linear gradient elution with a mobile phase containing methanol （1.96% acetic acid） and 10% methanol in water （1.96% acetic acid and 10 mmol/L ammonium acetate） at a flow rate of 0.2 mL/min. In addition, the major derivative, named derivative Ⅲ, was identified by UV spectra, MS, and the ^1H-NMR and ^13C-NMR spectra. Results： Good linearity was obtained within the range of 5 and 2000 ng/mL （r〉0.99 using a linear regression model with 1/x^2 weighting） for acetylshikonin. The interday and intraday precisions were found to be less than 12.3%, with the exception of the lowest concentration, which was less than 17.2%. The interday and intraday accuracies, which were between-3% and 0.6%, were also observed. After the administration of acetylshikonin （80 mg/kg, po） in macaque monkeys, the pharmacokinetic parameters were obtained through the non-compartmental analysis, where the area under the concentration- time curve to the last measurable concentration, the terminal elimination half- life, and the mean residual time were 615.4±206.5 ng·h/mL,12.3±1.6 h, and 10.2±0.7 h, respectively. Conclusion： The method was validated and applied to the quantitative determination and pharmacokinetic study of acetylshikonin in the blood samples of macaque monkeys.Aim： To develop and validate a novel precolumn derivatization method for the quantitative determination and pharmacokinetic application of acetylshikonin in macaque monkeys by liquid chromatography-electrospray ionization tandem mass spectrometry （LC-ESI-MS/MS）. Methods： 2-Mercaptoethanol was added to the blood sample as the derivatization reagent. The derivatization reaction formed 1-major derivation product, Which was well correlated with acetylshikonin. The acetylshikonin concentrations in the biological samples were calculated by quantitative determination of the major derivation product using LCESI-MS/MS. Separation was achieved using a C18 column （2 mm×50 mm, 5μm） at room temperature and a linear gradient elution with a mobile phase containing methanol （1.96% acetic acid） and 10% methanol in water （1.96% acetic acid and 10 mmol/L ammonium acetate） at a flow rate of 0.2 mL/min. In addition, the major derivative, named derivative Ⅲ, was identified by UV spectra, MS, and the ^1H-NMR and ^13C-NMR spectra. Results： Good linearity was obtained within the range of 5 and 2000 ng/mL （r〉0.99 using a linear regression model with 1/x^2 weighting） for acetylshikonin. The interday and intraday precisions were found to be less than 12.3%, with the exception of the lowest concentration, which was less than 17.2%. The interday and intraday accuracies, which were between-3% and 0.6%, were also observed. After the administration of acetylshikonin （80 mg/kg, po） in macaque monkeys, the pharmacokinetic parameters were obtained through the non-compartmental analysis, where the area under the concentration- time curve to the last measurable concentration, the terminal elimination half- life, and the mean residual time were 615.4±206.5 ng·h/mL,12.3±1.6 h, and 10.2±0.7 h, respectively. Conclusion： The method was validated and applied to the quantitative determination and pharmacokinetic study of acetylshikonin in the blood samples of macaque monkeys.

关 键 词：acetylshikonin macaque monkey blood liquid chromatography-electrospray ionizationtandem mass spectrometry 2-MERCAPTOETHANOL precolumn derivatization PHARMACOKINETIC 

分 类 号：R96[医药卫生—药理学]