摘要
目的：建立测定荷瘤裸鼠血浆中和厚朴酚脂质体的高效液相色谱-质谱联用（LC-MS/MS）法,并进行荷瘤裸鼠体内药代动力学研究。方法：采用YMC C18（100 mm×2.1 mm,1.7μm）色谱柱,流动相B为0.025%氨水乙腈,A为0.025%氨水,在线脱气,流速0.8 m L/min。梯度洗脱：0~2 min,B相85%~85%;2~4 min,B相85%~10%;4~6 min,B相10%~85%;6~8 min,B相85%~85%,柱温40℃。进样量10μL。质谱条件：离子源电喷雾电离源（ESI）,负离子电离模式,扫描方式多级反应监测（MRM）,监测离子对m/z和厚朴酚（265.0→223.0）和内标（253.0→225.0）。结果：血浆中无干扰测定的内源性物质,每个样品的分析时间为8 min;和厚朴酚在0.500~1000 ng/m L呈良好的线性关系,定量下限为0.500 ng/m L,日内、日间精密度RSD均小于15%,低、中、高3种浓度的提取回收率〉64.71%。稳定性实验中,在各种贮存条件下血浆中和厚朴酚均较稳定。结论：本方法操作简便,特异性强,灵敏度高,可用于和厚朴酚脂质体的药代动力学研究,以期为临床应用提供参考。
Objective：To develop and validate a liquid chromatography-tandem mass spectrometry（LC-MS/MS）method for the quantification of honokiol in nude mouse plasma and to study the pharmacokinetics of honokiol liposomes in nude mice.Methods：Honokiol and internal standard（chrysophanol） were separated on a YMC,C_（18） column（100 mm×2.1 mm,1.7 μm） by gradient elution using 0.025%ammonia in acetonitrile（B） and 0.025%ammonia in water（A） as the mobile phase.The flow rate was 0.8 mL/min and the column temperature was 40℃.The elution program was optimized as follows：0-2 min,B（85%~85%）;2-4 min,B（85%~10%）;4-6 min,B（l0%~85%）;6~8 min,B（85%~85%）,the injection volume was 10 μL.An API-4000 Q trap mass spectrometer equipped with an electrospray ionization（ESI） source was used as detector and was operated in negative ion mode.Multiple reaction monitoring（MRM） was performed and the m/z of ions selected for quantitation were 265.0→223.0（honokiol） and 253.0→225.0（chrysophanol）.Results：The chromatograms showed no endogenous interfering peaks in blank mouse plasma.Each analysis was completed within 8 min.The linear range of honokiol in plasma was0.500~1000 ng/mL.The lower limit of quantification was 0.500 ng/mL.The intrarun and inter-run relative standard deviations were less than 15%.The extract recoveries were more than 64.71%.In the stability studies,honokiol was found to be stable in plasma under various storage conditions.Conclusion：The method is sensitive,accurate and convenient for the pharmacokinetic study of honokiol liposomes,for providing reference of clinical application.
出处
《生物技术通讯》
CAS
2015年第6期809-814,共6页
Letters in Biotechnology
