摘要
目的:建立快速测定大鼠血浆中脱水葛根素浓度的液相色谱-串联质谱分析方法,并探讨脱水葛根素在大鼠体内的药代动力学。方法:血浆样品加入甲醇沉淀蛋白后,用LC-MS/MS进行测定分析。色谱柱为Zorbax SB-C18column(4.6 mm×150 mm I.D.5.0μm,Agilent,USA),流动相为甲醇(含10 mmol·L 1乙酸胺)-0.1%甲酸溶液(20:80,V/V),流速0.6 mL min 1,温度40°C。检测应用三重串联四级杆质谱仪的多重反应监测模式(MRM),选择的检测离子如下:脱水葛根素399.1→281.0,染料木素(内标)271.0→215.0。结果:在大鼠血浆样品中,脱水葛根素浓度在1.5 5400 ng mL 1之间线性关系良好,最低检测限为1.50 ng·mL 1。准确度在95.73%103.18%之间,精密度RSD在4.33%7.86%之间。结论:建立的LC-MS/MS联用方法简单准确,灵敏度高,适用于大鼠血浆中对脱水葛根素的药代动力学研究。
AIM： To establish a sensitive and rapid liquid chromatographic-tandem mass spectrometry （LC-MS/MS） method for the quantitative analysis of dehydrated puerarin in rat plasma, and its application for pharmacokinetic studies. METHODS： A plasma sample was pretreated by one-step protein precipitation by the addition of five volumes of methanol The chromatographic separation was achieved on a Zorbax SB-C18 column （4.6 mm ×150 mm I.D. 5.0 μm, Agilent, USA） at 40 ℃ at a flow rate of 0.6 mL.min-1 by an isocratic elution consisting of 10 mmol·L-1 ammonium acetate in methanol and water containing 0.1% formic acid in a ratio of 20 ： 80 （V/V）. Detection was performed on a triple quadrupole mass spectrometer in multiple-reaction monitoring （MRM） mode. An atmos- pheric pressure chemical ionization （APCI） interface in positive ionization mode was used by monitoring the transitions from m/z 399.1 →281.0 （dehydrated puerarin） and m/z 271.0→215.0 （internal standard, IS）. RESULTS： Calibration curves were linear in the concen- tration range from 1.50 to 5400 ng.mL-1, and the lower limit of quantification （LLOQ） was 1.50 ng.mL-1 in rat plasma. The accuracy and precision values, which were calculated from three different sets of quality control samples analyzed in sextuplicate on three dif- ferent days, ranged from 95.73% to 103.18%, and from 4.33% to 7.86%, respectively. CONCLUSION： The method was successfully applied to assess the pharmacokinetics of dehydrated puerarin after oral administration in rats.
出处
《中国天然药物》
SCIE
CAS
CSCD
2013年第5期566-571,共6页
基金
supported by the Natural Science Foundation of China(No.31101241)
the Natural Science Foundation of Shandong Province,China(No.Q2007C13)
the Shandong Administration of Traditional Chinese Medicine(No.2009-152)
the Independent Innovation Foundation of Shandong University(IIFSDU,No.2012TS163)~~
