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

What is it about?

In this paper, we report a pre-column derivatization method with 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate for the quantitative determination of amino acid composition in Ganirelix which is a synthetic decapeptide. This work is significant because among all chemical characterization tests for therapeutic peptides, one of the most significant and challenging aspects is to demonstrate comparable amino acid composition in terms of the relative molar ratio of each amino acid between a generic drug product and reference listed drug (RLD). In general, Edman degradation is a suitable analytical technique to identify amino acid sequence present in any peptide. However, this analytical technique is not applicable for Ganirelix is due to the acylation of N-terminal amino group. To address this challenge, a pre-column derivatization method was developed with 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent. In the reported method, an adequate chromatographic separation for all the amino acids of Ganirelix was achieved using c-18 reverse phase stationary phase. This method was applied successfully in the determination of amino acids in Ganirelix in order to demonstrate generic peptides that have the amino acid composition same as the reference listed drug.

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Photo by National Cancer Institute on Unsplash

Photo by National Cancer Institute on Unsplash

Why is it important?

Temperature and humidity levels can fluctuate significantly during transportation. Changes in the manufacturing process and storage environment are extremely sensitive to peptide molecules, resulting in modifications to their primary (amino acid composition and sequence) and secondary structures. Changes in the primary and secondary structure of certain peptides can have a significant impact on their biological function, so it is critical to evaluate their primary and secondary structural integrity after any processing or storage variation.