Department of Analytical Chemistry, Faculty of Chemical Engineering, UCT Prague, Technická 5, 166 28 Prague 6, Czech Republic.
Zentiva, k.s. Praha, U Kabelovny 130, 102 37 Prague 10, Czech Republic.
J Pharm Biomed Anal. 2022 May 10;213:114677. doi: 10.1016/j.jpba.2022.114677. Epub 2022 Feb 18.
The ultra-performance liquid chromatography (UPLC) method, which involves pre-column derivatization of nitrite with 2,3-diaminonaphthalene (DAN) to form 2,3-naphthotriazole (NAT), offers the advantages of easy sample preparation, simple derivatization, stable derivatives, rapid analysis, high sensitivity and specificity and lack of interferences for determining nitrite in pharmaceutical samples. Determination of NAT was performed on a an Acquity UPLC HSS T3 column using a gradient elution of 0.1% formic acid with acetonitrile at flow rate of 0.4 mL/min and temperature at 45 °C. The single-quadrupole mass detector was operated in the positive ion mode. Quadrupole mass analyser was employed in selected ion monitoring mode using a target ion at m/z = 170 as [M+H]. The UPLC-MS method was validated as per International Council on Harmonization (ICH) guidelines in terms of linearity, limit of detection, limit of quantification, selectivity, accuracy, precision, intermediate precision and stability. The UPLC-MS method was demonstrated to be applicable for the determination of nitrite in various pharmaceutical samples. The proposed UPLC-MS method was used to study the effect of nitrite content in pharmaceutical products on the formation of N-nitrosamines. The high importance of nitrites in relation to the N-nitrosation reaction was discussed. As deduced from theory and justified by the presented results, reducing the nitrite concentration could definitely solve the N-nitrosamine contamination. Nitrites, unlike secondary and tertiary amines, are universal precursors to any N-nitrosamine, so this solution is easily transferable to any relevant pharmaceutical product.
超高效液相色谱 (UPLC) 法，涉及亚硝酸根与 2,3-二氨基萘 (DAN) 预柱衍生化形成 2,3-萘三唑 (NAT)，具有样品制备简单、衍生化简单、衍生物稳定、分析快速、灵敏度和特异性高、干扰少等优点，用于测定药物样品中的亚硝酸根。NAT 的测定在 Acquity UPLC HSS T3 柱上进行，采用 0.1%甲酸和乙腈的梯度洗脱，流速为 0.4 mL/min，温度为 45°C。单四极杆质谱检测器以正离子模式工作。四极杆质谱仪在选择离子监测模式下使用目标离子 m/z = 170 作为 [M+H] 进行操作。UPLC-MS 方法按照国际协调会议 (ICH) 指南进行了验证，包括线性、检测限、定量限、选择性、准确性、精密度、中间精密度和稳定性。UPLC-MS 方法适用于测定各种药物样品中的亚硝酸根。该方法用于研究药物产品中亚硝酸根含量对 N-亚硝胺形成的影响。讨论了亚硝酸盐与 N-亚硝化反应的重要性。从理论推断和提出的结果来看，降低亚硝酸盐浓度肯定可以解决 N-亚硝胺污染问题。亚硝酸盐与仲胺和叔胺不同，是任何 N-亚硝胺的通用前体，因此该解决方案很容易转移到任何相关的药物产品中。
