Determination of Amino Acid Composition of Ganirelix Acetate in an Injectable Formulation by Pre-column Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate.

Journal of Chromatographic Science
Kumarswamy Ummiti, J V Shanmukha Kumar

Abstract

Ganirelix is a synthetic decapeptide linked with nine different amino acids. To understand the peptide amino acid sequence or primary structure, the first step is to determine the amino acid composition of the peptide which can be a determining factor for the peptide immunogenicity. Edman degradation is not a suitable analytical technique to identify amino acid sequence present in Ganirelix due to the absence of uncharged N-terminal amino group. To address this challenge, a pre-column derivatization method was developed with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent. In the present work, the Ganirelix active pharmaceutical ingredient present in the injectable formulation was isolated by fraction collection and further purified by flash chromatography. The amino acid composition of Ganirelix is assayed by carrying out acid hydrolysis with 6 mol L-1 hydrochloric acid solution containing 1% phenol at 100°C for 24 h and derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent solution, followed by determination of individual amino acids by reverse-phase chromatography using a C18 column. High resolution was achieved for the nine amino acid mixture. The amino acid composition results of temperature-s...Continue Reading

References

Mar 1, 1967·European Journal of Biochemistry·P Edman, G Begg
Mar 5, 2011·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ibolya Molnár-Perl
Apr 12, 2012·Journal of Assisted Reproduction and Genetics·Enis OzkayaKutluk Oktay
May 23, 2012·Systems Biology in Reproductive Medicine·Miriam HerediaJuan Carlos Valenzuela
Sep 18, 2012·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xiaoli ZhangHaixia Zhang
May 28, 2019·Reproductive Biomedicine Online·Serafeim PousiasKonstantinos Dafopoulos

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