Development and Validation of RP-HPLC Method for the Simultaneous Estimation of Paracetamol and Tramadol Hydrochloride in Tablet Dosage Form

Abstract

A simple reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been developed and
validated for simultaneous determination of Paracetamol and Tramadol hydrochloride in tablet dosage form.
Chromatographic analysis was performed on a Symmetry Thermo C18(150X4.6 mm,5µm) column ambient
temperature with a mixture of mixed phosphate buffer and Acetonitrile in the ratio 60:40 (mixed phosphate
buffer preparation; 0.01 M Potassium dihydrogen orthophosphate, pH 3.5 adjust with triethylamine) as mobile
phase, at a flow rate of 0.80 mL min-1
. UV detection was performed at 268 nm. The method was validated for
accuracy, precision, specificity, linearity and sensitivity. The retention times of Paracetamol and Tramadol
hydrochloride were 2.250 and 3.378 min, respectively. Calibration plots were linear over the concentration
ranges 62.500-375.000 μg mL-1
and 6.25-37.50 μg mL-1
for Paracetamol and Tramadol hydrochloride
respectively. The Limit of detection was 1.8704 and 1.254 µg mL-1
and the quantification limit was 5.6679 µg
mL-1
and 3.8008 µg mL-1 for Paracetamol and Tramadol hydrochloride respectively. The accuracy of the
proposed method was determined by recovery studies and found to be 99.56% to 100.55%.