A simple reversed-phase high-performance liquid chromatographic (RP-HPLC) technique for the simultaneous determination of ascorbic acid (ASC), methionine (MET), paracetamol (PAR) and caffeine (CAF) has been developed and validated. The cited components are separated completely using Brownlee Bio C18 column (250 x 4.6 mm, 5 μm) by isocratic elution of water-acetonitrile (85:15) (v/v) mobile phase flowing at 1.0 mL min-1
at ambient temperature. The spectrophotometric detection is carried out sequentially at 260 nm for ASC (2 min), 200 nm for MET (1 min), 240 nm for PAR (1.5 min) and 270 nm for CAF (1.5 min). Total chromatographic analysis time per samplewas approximately 6 min. The linear range of determination for ASC, MET, PAR and CAF are 40-160 μg mL-1, 40-200 μg mL-1, 20-400 μgmL-1 and 40-160 μg mL-1, respectively. Thus, proposed method can be successfully applicable to analysis the pharmaceutical preparation containing the above mentioned drugs without any interference of excipients. Recovery ranges and relative standard deviation are in turn 96.46 to 102.70 %, 2.65 % for ASC, 96.33 to 103.43 %, 2.93 % for MET, 98.31 to 102.73 %, 2.09 % for PAR and 95.82 to 102.13 %, 2.68% for CAF.
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