Abstract

AN ANALYTICAL NEW RP-HPLC METHOD FOR QUANTITATIVE ESTIMATION OF CLONAZEPAM IN BULK FORM AND MARKETED PHARMACEUTICAL TABLET DOSAGE FORM

M. Naresh*, A. Rajamani and A. Yasodha

ABSTRACT

Objective: To develop and validate a simple, selective, precise, and accurate method for the estimation of Clonazepam using Reversed-Phase – High Performance Liquid Chromatography (RP-HPLC) technique in Bulk and Marketed Formulation. Methods: The proposed method utilizes chromatographic conditions Phenomenex Luna (C18) RP Column, 250 mm x 4.6 mm, 5μm, mobile phase was Phosphate Buffer (pH-4.6) and Methanol in the proportion of 65:35% ratio, flow rate was maintained 1.0 ml/minute, column temperature was maintained at Ambient, detection wave length was 286 nm, and diluent was mobile phase. Results: By injecting 6 times of the standard solution system suitability parameters were studied and results were found well under the acceptance criteria. The linearity study was performed by taking 60-140% levels, and the R2 value was found to be 0.9995, precision was found to be 0.327 for repeatability and 0.767 & 1.34 for intermediate precision. The % recovery was found to be 100.437%. Limit of detection and limit of quantitation were found to be 1.469μg/ml and 4.454μg/ml, respectively. The % purity was found to be 99.398%. Degradation study on Clonazepam was performed and concluded that the purity threshold was more than purity angle and within the acceptable range. Conclusion: The developed RP-HPLC method for Clonazepam was found to be simple, precise, accurate, reproducible, and cost effective. Statistical analysis of the developed method conforms that the proposed method is an appropriate and it can be useful for the routine analysis. This method gives the basic idea to the researcher who is working in area such as product development and finish product testing.

Keywords: Clonazepam, High-Performance Liquid Chromatography, System Suitability.