摘要
目的建立测定老年人血浆中石杉碱甲的液相色谱-串联质谱方法。方法在血浆中加入内标石杉碱乙,用乙腈直接沉淀蛋白法处理样品。采用SeQuant ZIC-HILIC(100mm×2.1mm I.D,3.5μm)色谱柱进行分离,柱温35℃;流动相为含5 mmol·L^(-1)甲酸铵0.1%甲酸水溶液-乙腈(35:65,V/V),流速0.3 mL·min^(-1),柱温35℃,梯度洗脱,进样体积5μL;正离子多反应检测(MRM),离子通道分别为m/z 243.2→210.2(石杉碱甲),m/z 257.2→198.6(石杉碱乙)。结果石杉碱甲线性范围0.05～10μg·L^(-1)(r>0.999),定量下限0.05μg·L^(-1),提取回收率在81.0%～95.4%,批内、批间精密度RSD均小于8%,内标校正基质因子在2.11～2.28。主要药动学参数t_(max)为(2.2±0.7)h,ρ_(max)为(0.9±0.2)μg·L^(-1),t_(1/2)为(14.1±2.0)h,AUC_(0-∞)为(15.3±4.5)μg·h·L^(-1)。结论本方法操作简便、特异性强、灵敏度高,适用于石杉碱甲在老年人中的药动学研究。
AIM To establish a method for determination of huperzine A in human plasma by LC-MS/MS for elder population. METHODS After addition of huperzine B （internal standard）, the plasma was processed by direct protein precipitation. The chromatographic separation was performed on SeQuant ZIC- HILIC Column （ 100 mm x 2.1 mm I.D, 3.5 μm） column with a mobile phase of 5 mmol.L-1 ammonium formate and 0.1% formic acid solutionacetonitrile （35 ： 65, V/V） in water. Flow rate was 0.3 mL.min-1 and column temperature was kept at 35 ℃. Isocratic elution and the injection volume was 5 μL. The protonated ions of analytes weredetected in positive ionization through multiple reaction monitoring mode （MRM）. The mass transition pairs of m/z 243.2 - 210.2 and rrdz 257.2 - 198.6 were used to detect huperzine A and huperzine B respectively. RESULTS The linear concentration ranges of the calibration curve was 0.05 to 10 μg. L-1 and the low limit of quantitation was 0.05 μg L-1 for huperzine A （r 〉 0.999） . The extraction recovery rates were ranged from 81.0% to 95.4%, within-batch RSD and between-batch RSD were less than 8%, the matrix effect factor of 2.11 - 2.28. The main pharmacokinetics were as follows： （2.2 ± 0.7） h, p （0.9 ± 0.2） （14.1 ± 2.0） h and AUCo（15.3 ± 4.5） μg.h.L-L CONCLUSION simple and quick, with high specificity and sensitivety, suitable cokinetics in elder people. The pretreatment with LC-MS/MS method is for the research of huperzine A pharmacokinetics in elder people.
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2012年第12期728-731,共4页
Chinese Journal of New Drugs and Clinical Remedies
基金
中国科学院上海药物研究所临床研究中心基金项目(SIMMCRC2010001)
关键词
石杉碱甲
色谱法
高压液相
串联质谱法
血浆
老年人
agedhuperzine A
chromatography, high pressure liquid
tandem mass spectrometry
plasma
