Todmal Umesh, Suresh P S, Zainuddin Mohd, Kanth Bhamidipati Ravi, Samanta Swapan Kumar, Hallur Gurulingappa, Rajagopal Sridharan, Rajagopal Sriram, Mullangi Ramesh
Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Yeshwanthpur, Bangalore 560022, India.
Department of Medicinal Chemistry, Jubilant Biosys Ltd, Industrial Suburb, Yeshwanthpur, Bangalore 560022, India.
J Chromatogr Sci. 2017 Aug 1;55(7):750-756. doi: 10.1093/chromsci/bmx032.
A rapid and sensitive assay method has been developed and validated using liquid chromatography coupled to tandem mass spectrometry with electrospray ionization in the positive-ion mode for the estimation of SF0034 in mice plasma. The assay procedure involves a simple protein precipitation of SF0034 and tolbutamide (internal standard, IS) from mice plasma. Chromatographic separation was performed on an Atlantis dC18 column using an isocratic mobile phase comprising 0.2% formic acid:acetonitrile (10:90, v/v) at a flow rate of 0.60 mL/min. The total run time was 2.5 min. For mass spectrometric detection, the multiple reaction monitoring was used and ion transitions monitored were m/z 322 → 248 for SF0034 and 271 → 155 for IS. Method validation was performed as per regulatory guidelines and the results met the acceptance criteria. A calibration curve was constructed in the range of 2.08-2,078 ng/mL. The intra- and inter-day precision was in the range of 1.06-14.4% and 7.16-11.7%, respectively. This novel method has been applied to a pharmacokinetic study in mice.
已开发并验证了一种快速灵敏的检测方法，该方法采用液相色谱-串联质谱联用技术，以电喷雾电离正离子模式来测定小鼠血浆中的SF0034。检测程序包括从小鼠血浆中对SF0034和甲苯磺丁脲（内标，IS）进行简单的蛋白沉淀。色谱分离在Atlantis dC18柱上进行，使用等度流动相，该流动相由0.2%甲酸：乙腈（10:90，v/v）组成，流速为0.60 mL/min。总运行时间为2.5分钟。质谱检测采用多反应监测模式，监测的离子跃迁为SF0034的m/z 322 → 248和内标的m/z 271 → 155。按照监管指南进行方法验证，结果符合验收标准。在2.08 - 2,078 ng/mL范围内构建校准曲线。日内和日间精密度分别在1.06 - 14.4%和7.16 - 11.7%范围内。这种新方法已应用于小鼠的药代动力学研究。
