高效液相色谱-串联质谱法检测小鼠血浆和脑组织中沙奎那韦

摘要

本研究建立并验证了一种快速且灵敏的液质联用方法,用于测定小鼠血浆和脑组织中沙奎那韦的浓度,并将其应用于初步筛选实验,以评价黄酮类化合物对沙奎那韦脑内分布影响的作用.血浆和脑组织中的沙奎那韦和内标利托那韦通过液-液萃取的方式进行提取,色谱分离过程使用C18反相色谱柱(150mm×2.1 mm,5.0μm).质谱检测过程使用三重四极杆串联质谱,以电喷雾正离子模式、质谱多反应监测技术对沙奎那韦和内标进行检测.血浆中测定标准曲线的线性范围为0.1至10 ng/mL,其最低浓度为定量下限.方法日间日内精密度为7.5％-12.1％,准确度介于90.5％与107.2％之间.脑组织中测定标准曲线的线性范围为0.1至10 ng/g,其最低浓度为定量下限.方法日间日内精密度为7.3％-11.9％,准确度介于90.8％与107.4％之间.该方法成功应用于初步筛选实验,评价了19种黄酮类化合物对沙奎那韦脑内分布影响的作用,结果显示biochanin A能够最大程度促进沙奎那韦的脑内分布.%In the present study,a rapid and sensitive liquid chromatography-tandem mass spectrometric method for the determination of saquinavir in mice plasma and brain was developed,validated,and applied to a preliminary screening study evaluating the effect of bioflavonoids on the brain distribution of saquinavir in mice.Saquinavir and the intemal standard (ritonavir) were isolated from plasma and homogenized brain tissue matrices using a liquid-liquid extraction procedure,and the chromatographic separation was accomplished by using a reversed phase C18 column (150 mm×2.1 mm,5.0 μm).The analyte was detected by a triple-quadrupole tandem mass spectrometer via electrospray ionization,and multiple reaction monitoring was employed to select both saquinavir and ritonavir in the positive ion mode.A linear dynamic range of 0.1-10 ng/mL for plasma samples was established and the lower limit of quantification was 0.1 ng/mL.The intra-and inter-day precision were 7.5％-12.1％,and the accuracies ranged from 90.5％ to 107.2％ for plasma.A linear dynamic range of 0.1-10 ng/g for brain samples was established and the lower limit of quantification was 0.1 ng/g.The intra-and inter-day precision were 7.3％-11.9％,and the accuracies ranged from 90.8％ to 107.4％ for brain tissue samples.This method was successfully applied a preliminary screen of 19 bioflavonoids on the brain distribution of saquinavir in mice,and biochanin A shows the strongest effect.