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. 1999 Sep 3;855(1):147-55.
doi: 10.1016/s0021-9673(99)00669-x.

Separation of oxidized and deamidated human growth hormone variants by isocratic reversed-phase high-performance liquid chromatography

G Karlsson  1 , P Gellerfors, A Persson, B Norén, P O Edlund, C Sandberg, S Birnbaum
Affiliations

Separation of oxidized and deamidated human growth hormone variants by isocratic reversed-phase high-performance liquid chromatography

G Karlsson et al. J Chromatogr A. .

Abstract

Reversed-phase high-performance liquid chromatography (RP-HPLC) was utilized for the separation of recombinant human growth hormone (hGH) variants on a C18 silica column at 55 degrees C using an isocratic mobile phase which contained 27% 1-propanol in a 25 mM potassium phosphate buffer, pH 6.5. Three of the obtained peaks were characterized by tryptic mapping and mass spectrometry; two of the peaks were found to contain oxidized hGH (dioxy Met14/Met125 and Met125 sulfoxide) while the third contained a deamidated form (Asn149-->Asp149 or Asn152-->Asp152). Compared to the European Pharmacopoeia RP-HPLC method of hGH analysis, this new method gives two additional peaks and a 50% reduction in the analysis time.

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