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A Highly Validated RP-HPLC Method for Pyrazoline Derivative Having Anti-inflammatory Activity


Abstract

Author(s): Sivagami. B, Chandrasekar. R, Selvamani.P, Ramesh.R, Sudakar.P

A highly validated sensitive and specific reverse phase high performance liquid chromatography (RP-HPLC) method have been developed for the determination of synthesized product of pyrazoline derivative. The chromatographic separation was achieved using HPLC column Eclipse XDB C18 (150mm X 4.6mm X 5µm) column at isocratic mode. The mobile phase consists of 0.1% trifluoroacetic acid and methanol in the ratio of 20: 80. The flow rate and column temperature was maintained as 1.0 mL min-1 and 25 ± 2ºC respectively throughout the analysis. The injection volume was maintained as 5.0µL and the detection was carried out at 206 nm. The current method demonstrates good linearity over the range of 50-80 µg mL-1 and regression coefficient (r2 ) was found to be 0.998. The limit of detection (LOD) and limit of quantification (LOQ) of the compound was found to be 4 µg mL-1 and 15µg mL-1 respectively. The method was validated in accordance with ICH guidelines which include accuracy, precision, specificity, linearity, ruggedness, robustness, stability and system suitability. In addition, the current method has been utilized for quantification and routine analysis of other pyrazoline derivatives.

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