Abstract
An ultra high performance liquid chromatography-electrospray ionization-tandem mass spectrometric method (UHPLC/ESI-Q-TOF-MS) for the analysis of dopamine (DA) in Wistar rat brain homogenate has been developed and validated. The chromatographic separation was achieved on a Waters ACQUITY UPLC BEH C18 (100.0mmx2.1mm; 1.7 mu m) column using isocratic mobile phase, consisting of acetonitrile and Formic acid (0.1% w/v) (10: 90; v/v), at a flow rate of 0.15mlmin(-1). The transitions occurred at m/z 154.04137.006 for DA, and m/z 184.204166.08 for the internal standard. The recovery of the analytes from Wistar rat brain homogenate was optimized using liquid-liquid extraction technique (LLE) in ethyl acetate. The total run time was 3.5min and the elution of DA occurred at 1.44 +/- 0.05min. The linear dynamic range was established over the concentration range 75-750ngmL(-1) (r(2); 0.9921 +/- 0.0005) for DA. The intra-assay and inter-assay accuracy in terms of % CV was in the range 0.73-2.80. The lower limit of detection (LOD) and quantitation (LOQ) for DA was 0.278 and 0.844ngmL(-1), respectively. Analytes were stable under various conditions (in autosampler, during freeze-thaw, at room temperature, and under deep-freeze conditions). The developed method was successfully applied for in vivo profiling in rodents. Copyright (c) 2012 John Wiley & Sons, Ltd.