Analytical Science Development, Henlius Biologics Co., Ltd, 5155 Guangfulin Road, Shanghai 201616, China.
Analytical Science Development, Henlius Biologics Co., Ltd, 5155 Guangfulin Road, Shanghai 201616, China.
J Pharm Biomed Anal. 2022 Sep 5;218:114867. doi: 10.1016/j.jpba.2022.114867. Epub 2022 Jun 2.
Nitrosamine impurities are being detected in various pharmaceutical products recently. However, no analytical method is provided for biopharmaceuticals. In present work, a salting-out liquid-liquid extraction (SALLE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for quantification of thirteen nitrosamine contaminations in antibody drugs. The method showed excellent linearity over the range of 0.5-5.0 μg/L with LOQ (limit of quantitation) of 0.5 μg/L for targeted nitrosamines. The method was demonstrated to be accurate (recovery in a range of 75.4-114.7 %) and precise (RSD ≤ 13.2 %) for all nitrosamines using spiked samples. Especially, we found that the satisfactory recoveries for N-nitrosomethyl-4-aminobutyric acid (NMBA, 78.0-96.0 %) and 1-methyl-4-nitrosopiperazine (MeNP, 90.0-109.0 %) were just obtained in the opposite condition (with and without formic acid, respectively). In conclusion, we provide a sensitive and reliable method for nitrosamine estimations to ensure the safety of biological medications.
最近，各种药物产品中都检测到亚硝胺杂质。然而，目前还没有针对生物制药的分析方法。本工作建立了一种用于测定抗体药物中 13 种亚硝胺污染的盐析液液萃取（SALLE）-液相色谱-串联质谱（LC-MS/MS）方法。该方法在 0.5-5.0μg/L 范围内具有良好的线性关系，目标亚硝胺的定量限（LOQ）为 0.5μg/L。用加标样品验证了该方法对所有亚硝胺的准确性（回收率在 75.4-114.7%范围内）和精密度（RSD≤13.2%）。特别地，我们发现，在相反的条件下（分别有无甲酸），N-亚硝基甲基-4-氨基丁酸（NMBA，78.0-96.0%）和 1-甲基-4-亚硝基哌嗪（MeNP，90.0-109.0%）的回收率令人满意。总之，本研究为亚硝胺的测定提供了一种灵敏可靠的方法，以确保生物药物的安全性。
