Analytical Chemistry in Development and Supply, MMD, Merck & Co., Inc., 126 E. Lincoln Ave, Rahway, NJ 07065, USA.
Analytical Chemistry in Development and Supply, MMD, Merck & Co., Inc., 126 E. Lincoln Ave, Rahway, NJ 07065, USA.
J Chromatogr A. 2024 Nov 8;1736:465399. doi: 10.1016/j.chroma.2024.465399. Epub 2024 Sep 25.
Nitrosamine drug substance related impurities (NDSRIs) are often analyzed using high performance liquid chromatography (HPLC) with mass spectrometry (MS) detection. Due to high sensitivity requirements, high resolution MS or MS/MS is commonly used. However, it is difficult to implement this type of method for routine analysis at a supply site. Herein, we report a systematic approach to develop and validate a practical, robust, and user-friendly method for the analysis of NDSRIs using an inexpensive single quadrupole MS instrument such as QDa. We used 7-nitroso-3-(trifluoromethyl)-5,6,7,8-tetrahydro- [1,2,4] triazolo [4,3-a] pyrazine (NTTP) as an example to demonstrate the method development process. By optimizing the HPLC and MS parameters, we were able to develop a simple HPLC-MS method that provides the desired specificity and sensitivity for the analysis of NTTP and can be easily implemented in an analytical lab. The limit of quantitation is 0.5 ng/mL, corresponding to 0.1 ppm with respect to 5 mg/mL sitagliptin. The method has been successfully validated per ICH guidelines.
亚硝胺类药物杂质（NDSRIs）通常采用高效液相色谱（HPLC）与质谱（MS）联用进行分析。由于灵敏度要求较高，通常采用高分辨率 MS 或 MS/MS。然而，在供应现场进行常规分析时，很难实施这种方法。在此，我们报告了一种系统的方法，用于开发和验证一种实用、稳健且易于使用的方法，使用廉价的单四极杆 MS 仪器（如 QDa）分析 NDSRIs。我们以 7-亚硝基-3-（三氟甲基）-5,6,7,8-四氢-[1,2,4]三唑并[4,3-a]吡嗪（NTTP）为例，展示了方法开发过程。通过优化 HPLC 和 MS 参数，我们成功开发了一种简单的 HPLC-MS 方法，该方法对 NTTP 的分析具有所需的特异性和灵敏度，并且可以在分析实验室中轻松实施。定量限为 0.5ng/mL，相当于 5mg/mL 西他列汀中 0.1ppm。该方法已按照 ICH 指南成功验证。
