RP-HPLC with UV- Detection Method Development for the Simultaneous Quantification of Gabaenetin and Nortriptyline in Bulk and Tablet Dosage Form

Abstract

A Reverse phase high performance liquid chromatography with UV detection method is developed for the simultaneous quantification of Gabapenetin and Nortriptyline in bulk and in its combined tablet dosage form. The chromatographic separation was performed on YMC C8 column (150mm×4.6mmI.D 5 μm particle size) using isocratic elution.

The optimized mobile phase consists of 0.1% the orthophosphoric acid and methanol (60:40 v/v). The eluted analytes are monitored at 238 nm wavelength using a UV detector. The developed method separates Gabapenetin and Nortriptyline 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. Linearity was observed in concentration range of 200-600 µg/ml for Gabapenetin and 5-15 µg/ml for Nortriptyline. The LOD was found to be 0.672 and 0.0117μg/ml for Gabapenetin and Nortriptyline respectively. The LOQ was found to be 2.240 and 0.0390 μg/ml for Gabapenetin and Nortriptyline, respectively. The range of percentage recovery of Gabapenetin and Nortriptyline was found to be 100.49-100.75% and 100.35-100.51%, respectively. The % RSD values of precision study for Gabapenetin were found to be 0.227% and for Nortriptyline it was foundtobe0.347%. All the validation parameter values are within the acceptable range. The developed and validated method was successfully applied to the determination of Gabapenetin and Nortriptyline in combined pharmaceutical dosage form without any interference from the excipients with good recovery, precision and accuracy.