Soyseven Murat, Keçili Rüstem, Aboul-Enein Hassan Y, Arli Göksel
Department of Medical Services and Techniques, Yunus Emre Vocational School of Health Services, Anadolu University, Eskişehir 26470, Turkey.
Department of Pharmacy, Program in Pharmacy Services, Yunus Emre Vocational School of Health Services, Anadolu University, Eskişehir 26470, Turkey.
J Chromatogr Sci. 2021 Feb 15;59(3):241-245. doi: 10.1093/chromsci/bmaa100.
A novel analytical method, based on high-performance liquid chromatography with a UV (HPLC-UV) detection system for the sensitive detection of a genotoxic impurity (GTI) 5-amino-2-chloropyridine (5A2Cl) in a model active pharmaceutical ingredient (API) tenoxicam (TNX), has been developed and validated. The HPLC-UV method was used for the determination of GTI 5A2Cl in API TNX. The compounds were separated using a mobile phase composed of water (pH 3 adjusted with orthophosphoric acid): MeOH, (50:50: v/v) on a C18 column (150 × 4.6 mm i.d., 2.7 μm) at a flow rate of 0.7 mL min-1. Detection was carried out in the 254 nm wavelength. Column temperature was maintained at 40°C during the analyses and 10 μL volume was injected into the HPLC-UV system. The method was validated in the range of 1-40 μg mL-1. The obtained calibration curves for the GTI compound was found linear with equation, y = 40766x - 1125,6 (R2 = 0.999). The developed analytical method toward the target compounds was accurate, and the achieved limit of detection and limit of quantification values for the target compound 5A2Cl were 0.015 and 0.048 μg mL-1, respectively. The recovery values were calculated and found to be between 98.80 and 100.03%. The developed RP-HPLC-UV analytical method in this research is accurate, precise, rapid, simple and appropriate for the sensitive analysis of target GTI 5A2Cl in model API TNX.
已开发并验证了一种基于高效液相色谱-紫外（HPLC-UV）检测系统的新型分析方法，用于灵敏检测模型活性药物成分（API）替诺昔康（TNX）中的遗传毒性杂质（GTI）5-氨基-2-氯吡啶（5A2Cl）。HPLC-UV方法用于测定API替诺昔康中的GTI 5A2Cl。使用由水（用正磷酸调节pH至3）：甲醇（50:50，v/v）组成的流动相，在C18柱（内径150×4.6 mm，粒径2.7μm）上以0.7 mL min-1的流速分离化合物。在254 nm波长处进行检测。分析过程中柱温保持在40°C，将10μL体积的样品注入HPLC-UV系统。该方法在1-40μg mL-1范围内进行了验证。发现GTI化合物的校准曲线呈线性，方程为y = 40766x - 1125.6（R2 = 0.999）。所开发的针对目标化合物的分析方法准确，目标化合物5A2Cl的检测限和定量限分别为0.015和0.048μg mL-1。计算回收率，发现回收率在98.80%至100.03%之间。本研究中开发的RP-HPLC-UV分析方法准确、精密、快速、简单，适用于模型API替诺昔康中目标GTI 5A2Cl的灵敏分析。
