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Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-HPLC METHOD FOR ESTIMATION OF NEOSTIGMINE METHYL SULFATE INJECTION

Nikas Ravindra Ananta*, Prof. T. V. Chorage, Maind Pawan and Pote Vishal

ABSTRACT

The current proposed methodology of the research is to determine the related substances present in Neostigmine by using high-performance liquid chromatographic method. the developed method was validated for their accuracy and reproducibility. Reversed-phase chromatography was performed on Waters 2489 UV 2695 pump, Waters 2998 PDA 2695 pump Software Empower2 photodiode array detector using Ace C18 (250 mm × 4.6 mm, 5 μm particle size) column with pH 6.4 buffer: methanol : acetonitrile in the ratio of 75:10:15 as mobile phase at a flow rate of 1.0 mL/min. by isocratic elution with UV detection at 220 nm. Recovery and Linearity was observed well within the limits (R2 = more than 0.99 for concentration range of LOQ to 150% level for linearity and the % recovery was within the ICH acceptance limits of 85-115%) for all the impurities. The limit of quantitation (LOQ) and limit of detection (LOD) were found to be less than 0.05%. The method was validated as per ICH guidelines. The RSD for intra-day and inter-day (<3.0% RSD) precision were found to be less than 1 %. The percentage recovery was in good agreement with the labeled amount in the pharmaceutical formulations. From the method validation data, it can be concluded that the method is simple, specific, precise and accurate for the determination of Neostigmine in pharmaceutical formulations.

Keywords: Neostigmine, Estimation of related substances, HPLC.


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