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-stressed Ganirelix generic product and reference listed drug are in good agreement with the theoretical molar ratio of label information.

中文翻译：

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用6-氨基喹啉基-N-羟基琥珀酰亚胺基氨基甲酸酯进行柱前衍生，测定注射用制剂中加尼利克斯乙酸酯的氨基酸组成。

Ganirelix是与九种不同氨基酸连接的合成十肽。为了理解肽的氨基酸序列或一级结构，第一步是确定肽的氨基酸组成，其可以是肽免疫原性的决定因素。由于缺乏不带电的N端氨基，Edman降解不适用于鉴定Ganirelix中存在的氨基酸序列的合适分析技术。为了解决这一挑战，开发了一种使用6-氨基喹啉基-N-羟基琥珀酰亚胺基氨基甲酸酯试剂的柱前衍生方法。在目前的工作中，通过级分收集分离存在于注射制剂中的Ganirelix活性药物成分，并通过快速色谱法进一步纯化。含有1％苯酚的^-1盐酸溶液在100°C下放置24小时，然后用6-氨基喹啉基-N-羟基琥珀酰亚胺基氨基甲酸酯试剂溶液进行衍生化，然后使用C18柱通过反相色谱法测定各个氨基酸。九种氨基酸的混合物达到了高分辨率。温度应激的Ganirelix通用产品和参考上市药物的氨基酸组成结果与标记信息的理论摩尔比非常吻合。