Development and Validation of HPLC Method for the Simultaneous Quantification of Acetaminophen and Tramadol in Combined Pharmaceutical Dosage Forms with UV Detection.

ABSTRACT

A new high performance liquid chromatography (HPLC) with ultraviolet detection method is developed for the simultaneous quantification of acetaminophen and tramadol in bulk and in its combined pharmaceutical dosage form. The chromatographic separation was performed on Waters symmetry C8 column (250 mm × 4.6 mm I.D., 5 μm particle size) using isocratic elution. The optimized mobile phase consists of phosphate buffer (pH 6.8) and methanol (8020, v/v). The eluted analytes are monitored at 215 nm wavelength using a UV detector. The developed method separates acetaminophen and tramadol within a run time of 6 min. The developed method was validated as per International Conference of Harmonization guidelines with respect to linearity, sensitivity (limit of detection and limit of quantification), selectivity, accuracy, precision and robustness. The developed and validated method was successfully applied to the determination of acetaminophen and tramadol in combined pharmaceutical dosage forms without any interference from the excipients with good recovery, precision and accuracy.