Division of Chemical Metrology & Analytical Science, National Institute of Metrology, Beijing 100029, PR China.
Division of Chemical Metrology & Analytical Science, National Institute of Metrology, Beijing 100029, PR China.
J Pharm Biomed Anal. 2020 Jul 15;186:113271. doi: 10.1016/j.jpba.2020.113271. Epub 2020 Mar 20.
Calcitonin salmon is an important peptide pharmaceutical, which is mainly used for the treatment of osteoporosis and hypercalcemia. Structurally related peptide impurities in a peptide pharmaceutical probably have side effect or even toxicity, thus needs to be carefully characterized according to pharmacopoeia. With the improvement of analytical techniques, liquid chromatography-high resolution mass spectrometry (LC-HRMS) has become a pivotal technique for the identification and quantification of structurally related peptide impurities in peptide materials. In this study, an LC-HRMS-based method has been developed for the identification and quantification of structurally related peptide impurities in calcitonin salmon material. With this method, 7 peptide impurities (> 1 mg/g) in United States Pharmacopoeia (USP) reference standard and 9 peptide impurities (> 1 mg/g) in European Pharmacopoeia (EP) reference standard were identified and accurately quantified. Besides the peptide impurities reported by USP and EP, several new impurities such as [7-Dehydroalanine] calcitonin salmon, triple-sulfate-calcitonin salmon, [26-Proline] calcitonin salmon, [14-Glutamic acid] calcitonin salmon, [20-Glutamic acid] calcitonin salmon, [26-Aspartic acid] calcitonin salmon, calcitonin salmon acid were observed in the reference standard materials studied. The total mass fractions of all structurally related peptide impurities in calcitonin salmon study materials were estimated to be 57.4 mg/g for USP and 46.3 mg/g for EP with associated expended uncertainties at a 95 % confidence level of 5.2 mg/g (k = 2) and 3.1 mg/g (k = 2), respectively.
鲑鱼降钙素是一种重要的肽类药物，主要用于治疗骨质疏松症和高钙血症。肽类药物中结构相关的肽类杂质可能具有副作用，甚至毒性，因此需要根据药典进行仔细的特征描述。随着分析技术的提高，液相色谱-高分辨质谱（LC-HRMS）已成为鉴定和定量肽类物质中结构相关肽类杂质的关键技术。本研究建立了一种基于 LC-HRMS 的方法，用于鉴定和定量鲑鱼降钙素原料中的结构相关肽类杂质。该方法鉴定并准确定量了美国药典（USP）参比标准品中的 7 种（>1mg/g）肽杂质和欧洲药典（EP）参比标准品中的 9 种（>1mg/g）肽杂质。除 USP 和 EP 报道的肽杂质外，还在研究的参比标准物质中观察到几种新的杂质，如[7-脱氢丙氨酸]鲑鱼降钙素、三硫酸盐鲑鱼降钙素、[26-脯氨酸]鲑鱼降钙素、[14-谷氨酸]鲑鱼降钙素、[20-谷氨酸]鲑鱼降钙素、[26-天冬氨酸]鲑鱼降钙素、鲑鱼降钙素酸。USP 鲑鱼降钙素研究材料中所有结构相关肽类杂质的总质量分数估计为 57.4mg/g，EP 鲑鱼降钙素研究材料中所有结构相关肽类杂质的总质量分数估计为 46.3mg/g，在 95%置信水平下，相应的扩展不确定度分别为 5.2mg/g（k=2）和 3.1mg/g（k=2）。
