A rapid UPLC-UV method development and validation for the quantitative determination of Formaldehyde using derivatization technique

Abstract

The Objective of the current study was to develop and validate a Rapid UPLC-UV method for the quantitation of Formaldehyde. Formaldehyde (HCHO) is reacted with 2,4-dinitrophenylhydrazine (DNPH) to form a Schiff base (HCHO-DNPH derivatization product). The chromatographic conditions were developed and optimized using a mixture of DNPH reagent and HCHO-DNPH derivatization product. The chromatographic separation was achieved on Acquity BEH C8, 100 mm x 2.1 mm, 1.7μ particle size column. Using Water and Acetonitrile (55:45, v/v) as a mobile phase with 0.4 mL/min flow rate in Isocratic mode, the column temperature was maintained at 35°C, detection wavelength was set at 360 nm and the injection volume was 5 μL. Acetonitrile was used as a diluent. The developed RP-UPLC method was validated according to ICH guidelines. In this method the LOD and LOQ values for Formaldehyde are 0.6 ppm and 2.0 ppm respectively. The percentage recovery was 96.3 to 97.0. The solution was observed to be stable up to 48 h at room temperature. The validated method produced good results of precision, linearity, accuracy, robustness and ruggedness. The proposed method was found to be Rapid and suitable for quantification of Formaldehyde which can be further extended in Drug product and its excipients analysis.