Abstract

RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE QUANTITATIVE DETERMINATION OF POTENTIAL IMPURITIES OF NEFOPAM

Bharathi Tejas G. J.* and Bhadre Gowda D. G.

Abstract

The objective of the study was to develop and evaluate the reverse phase liquid chromatography (RP-HPLC) method for the quantitative determination of potential impurities of Nefopam active pharmaceutical ingredient. The method uses a Puratis RP-18 column (250 × 4.6mm, 5μm) with mobile phase A consisted, 0.1% trifluoro acetic acid in water and mobile phase B consisted of acetonitrile with a gradient programme. The column temperature was maintained at 25 °C and the detection was carried out at 220 nm. Efficient and reproducible chromatographic separation was achieved on Puratis RP-18 stationary phase in gradient elution profile. The newly developed HPLC method was validated according to ICH guidelines considering four impurities to demonstrate precision, linearity, accuracy and robustness of the method. The developed HPLC method was found to be accurate and sensitive. The correlation coefficient values are greater than 0.99 for Nefopam and its four impurities. Detection limit and quantitation limit was 0.40ppm and 1.23ppm respectively, indicating the high sensitivity of the newly developed method. Accuracy of the method was established based on the recovery obtained between 99.78% and 102.07% for all impurities. The result of robustness study also indicates that the method is robust and is unaffected by small variation in chromatographic conditions. The proposed HPLC method provides reliable, reproducible, accurate and sensitive for the quantification of Nefopam related substances.

Keywords: Nefopam; Impurities; RP-HPLC; Validation.