Research Article
Sujan Kumar D. P.1, Palavan C.
Abstract
The quality of sample extraction had a significant impact on mass spectrometry results. The presence of phospholipids in the sample extracts resulted in poor quantitation and also it decreases the method robustness. Here we adopted a novel sample preparation Hybrid SPE phospholipid technology to extract plasma samples for improved phospholipid removal. This new method allowed simultaneous quantification of propafenone and 5-OH propafenone at lower levels 0.5 and 0.25 ng/mL respectively. The phospholipid free filtrate obtained through Hybrid SPE-Phospholipid cartridge was chromatographed onto Gemini C18 column (75 x 4.6 mm, 3.0 μm). An isocratic mobile phase of a mixture of 10mM ammonium formate (pH 3.0 adjusted with formic acid) and methanol (20:80%V/V) at a flow rate of 0.5 mL/min was used. Precursor ion and product ion transition for analytes and IS were monitored on a TSQ Vantage triple quadrupole mass spectrometer, operated in the positive ionization mode. Method was validated over a concentration range of 0.50-500.00 ng/mL for propafenone, 0.25-250.00 ng/mL for 5- OH propafenone. The intra- and inter-day precision over the concentration range for propafenone and 5-OH propafenone were lower than 6.1 and 14.2% (coefficient of variation, %CV), and accuracy was between 99.5–108.7 and 94.6–108.3%, respectively. By using this new Hybrid SPE-Phospholipid technology the risk of phospholipid accumulation on column was knocked out completely and resulted in good peak shape with excellent column performance.