Rezk Mamdouh R, Bendas Ehab R, Basalious Emad B, Karim Iman A
Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562, Cairo, Egypt.
Clinical Pharmacy Department, Faculty of Pharmaceutical Sciences & Pharmaceutical Industries, Future University in Egypt, Egypt.
J Pharm Biomed Anal. 2016 Sep 5;128:61-66. doi: 10.1016/j.jpba.2016.05.016. Epub 2016 May 11.
A rapid and sensitive UPLC-MS/MS method was developed and validated for determination of daclatasvir (DAC) in human plasma using sofosbuvir (SOF) as an internal standard (IS). The Xevo TQD LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. Precipitation with acetonitrile was used in sample preparation. The prepared samples were chromatographed on Acquity UPLC HSS C18 (50×2.1mm, 1.8μm) column by pumping 10mM ammonium formate (pH 3.5) and acetonitrile in an isocratic mode at a flow rate of 0.30ml/min. Method validation was performed as per the FDA guidelines and the standard curves were found to be linear in the range of 5-4000ng/ml for DAC. The intra-day and inter-day precision and accuracy results were within the acceptable limits. A very short run time of 1.2min made it possible to analyze more than 500 human plasma samples per day. The wider range of quantification of DAC allowed the applicability of the developed method for its determination in a bioequivalence study in human volunteers.
建立了一种快速灵敏的超高效液相色谱-串联质谱（UPLC-MS/MS）方法，并以索磷布韦（SOF）作为内标（IS）对人血浆中的达卡他韦（DAC）进行测定及方法验证。Xevo TQD LC-MS/MS采用电喷雾电离在多反应监测模式下运行。样品制备采用乙腈沉淀法。制备好的样品在Acquity UPLC HSS C18（50×2.1mm，1.8μm）色谱柱上进行分离，以10mM甲酸铵（pH 3.5）和乙腈等度洗脱，流速为0.30ml/min。按照美国食品药品监督管理局（FDA）指南进行方法验证，发现DAC的标准曲线在5 - 4000ng/ml范围内呈线性。日内和日间精密度及准确度结果均在可接受范围内。1.2分钟的极短运行时间使得每天能够分析500多个人类血浆样本。DAC更宽的定量范围使得所建立的方法适用于在人类志愿者中进行的生物等效性研究中的测定。
