Author/Authors :
Varcheh, Nastaran Nafissi Department of Pharmaceutics, School of Pharmacy, Shaheed Beheshti University of Medical Sciences and Health Services, Tehran , Aboofazeli, Reza Department of Pharmaceutics, School of Pharmacy, Shaheed Beheshti University of Medical Sciences and Health Services, Tehran , Shafaati, Alireza Department of Pharmaceutical Chemistry, School of Pharmacy, Shaheed Beheshti University of Medical Sciences and Health Services, Tehran , Zarghi, Afshin Department of Pharmaceutical Chemistry, School of Pharmacy, Shaheed Beheshti University of Medical Sciences and Health Services, Tehran
Abstract :
The accurate prediction of protein stability is one of the most challenging goals in protein
formulation and delivery. In this study, a gradient RP-HPLC method is described for the
separation of human growth hormone (hGH) variants as deamidated and oxidized forms. The
methodology employed a polymeric poly (styrene-co-divinylbenzene) column and a 1mL/min
flow rate of a linear gradient of 0.1% v/v TFA/acetonitrile and TFA/Water (pH=2.0) mixture
as the mobile phase. The overall run time of this method was 12 min and the average retention
times were about 8.7 min for the native somatropin, 7.2 min for the deamidated form and 1.6
and 5.3 min for oxidized variants. The method was also validated in terms of selectivity,
linearity, intra- and inter-day variations.
In conclusion the method was found to have the potential for being applied as an initial and
rapid evaluation method for assessing the quality and quantity of hGH during downstream
processing, formulation and storage
Keywords :
Somatropin , Degradation products , HPLC , Gradient , Polymeric column
