HPTLC method development for the simultaneous determination of Pregabalin and Amitryptyline hydrochloride in pharmaceutical dosage forms
A simple, selective, precise high performance thin layer chromatography (HPTLC) method with densitometry at λ= 205 nm was developed and validated for simultaneous determination of Pregabalin and Amitryptyline Hydrochloride in pharmaceutical dosage form. Chromatographic separation of the drugs were performed on aluminum plates precoated with silica gel 60 F254 used as stationary phase and the chromatogram was developed using Toluene: Methanol: Formic acid (7: 2.5: 0.5 v/v/v) and 20 ml of mobile phase was used per chromatography run. The system was found to give a compact band for Pregabalin (Rf =0.27±0.03) and Amitryptyline Hydrochloride (Rf =0.68±0.03). The validated lowest limit of detection was 45.097ng/spot and 12.614ng/spot wherenas lowest limit of quantification was 136.659ng/spot and 38.224ng/spot for Pregabalin and Amitryptyline Hydrochloride respectively. The percentage recovery for Pregabalin was found to be 99.91 (at 50%), 99.39 (at 100%), 99.27(at 150%) and 100.42 (at 50%), 100.63 (at 100%), 100.97(at 150%) for Amitryptyline Hydrochloride. Statistical analysis proved that the method is selective, precise and accurate for the estimation of Pregabalin and Amitryptyline Hydrochloride.
Keywords: Pregabalin (PRGB), Amitryptyline Hydrochloride (AMTR), HPTLC, Pharmaceutical formulation.
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