Nirogi Ramakrishna, Kandikere Vishwottam, Komarneni Prashanth, Aleti Raghupathi, Padala Nagasuryaprakash, Kalaikadhiban Ilayaraja
Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Road 5 Avenue 7, Banjara Hills, Hyderabad, India.
Biomed Chromatogr. 2012 Oct;26(10):1150-6. doi: 10.1002/bmc.2670. Epub 2012 Jan 4.
A sensitive high-performance liquid chromatography-positive ion electrospray tandem mass spectrometry method was developed and validated for the quantification of ambrisentan in plasma. The analyte and the internal standard (armodafinil) were extracted from plasma by acetonitrile precipitation and they were separated on a reversed-phase C(18) column with a gradient program. The MS acquisition was performed with multiple reaction monitoring mode using the respective M + H ions, m/z 379-347 for ambrisentan and m/z 274-167 for the IS. The assay exhibited a linear dynamic range of 1-2000 ng/mL for ambrisentan in plasma. Acceptable precision (<10%) and accuracy (100 ± 8%) were obtained for concentrations over the standard curve range. The method was successfully applied to quantify ambrisentan concentrations in a rodent pharmacokinetic study after a single oral administration of ambrisentan at 2.5 mg/kg to rats. Following oral administration the maximum mean concentration in plasma (C(max) ; 1197 ± 179 ng/mL) was achieved at 1.0 ± 0.9 h (T(max) ), and the area under the curve (AUC) was 6013 ± 997 ng h/mL. Therefore, development of such a simple and sensitive method in rat plasma should translate into a method for ambrisentan in human plasma for clinical trials.
建立了一种灵敏的高效液相色谱-正离子电喷雾串联质谱法，并对其进行了验证，用于定量测定血浆中的安立生坦。通过乙腈沉淀从血浆中提取分析物和内标（阿莫达非尼），并在反相C(18)柱上采用梯度洗脱程序进行分离。采用多反应监测模式，分别以各自的M + H离子进行质谱采集，安立生坦的m/z为379-347，内标的m/z为274-167。该测定法在血浆中安立生坦的线性动态范围为1-2000 ng/mL。在标准曲线范围内的浓度下，获得了可接受的精密度（<10%）和准确度（100 ± 8%）。该方法成功应用于在大鼠单次口服2.5 mg/kg安立生坦后的啮齿动物药代动力学研究中定量测定安立生坦浓度。口服给药后，在1.0 ± 0.9 h（T(max)）达到血浆中最大平均浓度（C(max)；1197 ± 179 ng/mL），曲线下面积（AUC）为6013 ± 997 ng h/mL。因此，在大鼠血浆中开发这样一种简单灵敏的方法应可转化为用于临床试验的人血浆中安立生坦的方法。
