Rapid Simultaneous Determination of Paracetamol, Amantadine Hydrochloride, Caffeine and Chlorpheniramine Maleate in Human Plasma by Liquid Chromatography/Tandem Mass Spectrometry

A rapid, simple and sensitive high performance liquid chromatography with positive ion electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) was first developed and validated to simultaneously determine paracetamol (PAR, CAS 103-90-2), amantadine hydrochloride (ATH, CAS 665-66-7), caffeine (CAF, CAS 58-08-2) and chlorpheniramine maleate (CPM, CAS 113-92-8) in human plasma using tramadol hydrochloride (TMH, CAS 22204-88-2) as internal standard (IS). Following methanol-induced protein precipitation, the analytes were separated using a mobile phase comprised of methanol: water (0.5% formic acid) = 20:80 (v/v) on a commercially available column (150 mm x 2.1 mm I.D., 5 mu m) and analyzed by electrospray ionization tandem mass spectrometry in the selected reaction monitoring (SRM) mode with the precursor to product ion transitions m/z 152.3 -> 110.2 for PAR, 152.3 -> 135.3 for ATH, 195.1 -> 138.3 for CAF, 275.2 -> 230.3 for CPM and 264.2 -> 58.2 for TMH. The standard curves were linear (r(2) > 0.99) over the concentration range of 0.2-20 mu g/mL for PAR, 20-2000 ng/mL for ATH and CAF, 0.1-10 ng/mL for CPM and had good accuracy and precision, respectively. The within- and between-batch precisions were less than 15% in terms of relative standard deviation (RSD). The lower limit of quantitation (LLOQ) were 0.2 mu g/mL, 20 ng/mL, 20 ng/mL and 0.1 ng/mL for PAR, ATH, CAT and CPM, respectively. The described method has been successfully applied to study the pharmacokinetics of paracetamol-amantadine hydrochloride tablets in Chinese healthy male volunteers with great precision and sensitivity.