This study aimed to develop an analytical method to determine the quantity of the impurity 3‐aminopyridine (3AP). 3‐Aminopyridine is a reactive reagent in the synthesis of linagliptin. The method was sensitive at level of 30.0 ppm of 3AP relative to linagliptin. The analysis was carried out using hydrophilic interaction liquid chromatography. The analytical column was Tracer Extrasil Silica (150 × 4.0 mm, 3 μm). A mobile phase of water–acetonitrile (10:90, v/v) containing 10.0 mM ammonium acetate was prepared and adjusted to pH 6.0. A UV detector was used to detect the amount of 3AP at a wavelength of 298 nm. Validation of the method was performed as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use in terms of detection limit, quantitation limit, linearity, accuracy, precision, specificity and robustness. The calibration curve was linear (r² = 0.999) for 3AP concentration in the range of 30.0–450.0 ppm. This method showed a good sensitivity with a detection limit and a quantitation limit of 7.5 and 25.0 ppm, respectively.

中文翻译：

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使用HILIC-UV定量测定利格列汀活性药物成分中潜在的遗传毒性杂质3-氨基吡啶。

这项研究旨在开发一种分析方法，以确定杂质3-氨基吡啶（3AP）的量。3-氨基吡啶是利格列汀合成中的一种反应试剂。相对于利格列汀，该方法在3AP ppm的30.0 ppm浓度下敏感。使用亲水相互作用液相色谱法进行分析。分析柱为示踪剂Extrasil Silica（150×4.0 mm，3μm）。制备了含有10.0 mM乙酸铵的水-乙腈流动相（10：90，v / v），并将其pH调节至6.0。使用UV检测器检测298nm波长下的3AP量。方法的验证是按照国际人用药品技术要求协调委员会的要求进行的，包括检测限，定量限，线性，准确性，精密度，特异性和鲁棒性。校正曲线为线性（3AP浓度在30.0–450.0 ppm范围内，r ² = 0.999）。该方法显示出良好的灵敏度，检测限和定量限分别为7.5和25.0 ppm。