Abstract

ESTIMATION OF RECOMBINANT HUMAN PARA THYROID HARMON (RH-PTH) AND ITS RELATED SUBSTANCES IN DRUG PRODUCT BY REVERSE PHASE HIGH PRESSURE LIQUID CHROMATOGRAPHY

Munnelli Pavani*, Karavadi Thejomoorthy, Ch. Saibabu

ABSTRACT

The aim of the present work was to develop and validate a simple, efficient, economical method for the estimation of Recombinant Human Para Thyroid Harmon (rh-PTH) and its related substances in Drug product by reverse phase high pressure liquid chromatography. Chromatography was performed on Dionex, Phenomenex C18 (250×4.6 mm, 5.0μm) with mobile phase containing 0.1%TFA in WFI and 0.1%TFA in 90% Acetonitrile at a flow rate of 1 mL/min and eluents were monitored at 280 nm The retention times of rh-PTH & m-Cresole were 11.0, and 10.33 min respectively and showed a good linearity in the concentration range of rh-PTH 100-300μg/mL for m-Cresol 0.5-1.5μg/mL with a correlation coefficient of 0.999 and 0.999 respectively. The validation characteristics included specificity, linearity, and limit of detection, limit of quantification, precision, robustness and stability. Validation acceptance criteria were met in all cases. The percent recoveries ranged between 95-105%, RSD < 2%. The method could be successfully used for the analysis of rh-PTH and its related substances in Drug Product.

Keywords: Recombinant Human Para Thyroid Harmone (rh-PTH), m-Cresol, Trifluro acetic acid, Acetonitrile.