A validated RP-HPLC
method for quantitative determination of genotoxic impurity hydrazine content in
Ursodeoxycholic acid
Ramya Sudha P., Bharath P. and Ramachandran D.
Res. J. Chem. Environ.; Vol. 29(5); 160-168;
doi: https://doi.org/10.25303/295rjce1600168; (2025)
Abstract
A highly sensitive method for the determination of genotoxic impurity such as hydrazine
in Ursodeoxycholic acid using RP-HPLC has been presented in the present study. Quantification
of hydrazine content in Ursodeoxycholic acid sample by HPLC was done with UV Detector.
Hydrazine was UV inactive compound. Derivatization procedure was established to
detect the hydrazine in HPLC. For this, 4-Hydroxy 3-methoxybenzaldehyde was used
as a derivatizing agent which reacts with hydrazine in the presence of disodium
tetraborate to form a compound which was UV active. Hydrazine was determined by
RP-HPLLC method using LiChrospher 100-RP18 (250 x 4.6mm, 5 μm) column as stationary
phase.
Column temperature maintained 35°C, injection volume 100μL, flow rate was 1.0 mL/min.
Hydrazine was detected using UV detector at the wavelength of 350 nm and run time
was 45 minutes. The mobile phase used water and methanol in gradient mode. The method
validation has been carried as per International Conference on Harmonization guidelines
(ICH). Limit of quantitation (LOQ) was found 0.21 ppm for hydrazine.
