Zheng Xiangyuan, Luo Lan, Zhou Jie, Ruan Xiaoling, Liu Wenyuan, Zheng Feng
Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, China; Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, China.
Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, China; Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, China.
J Pharm Biomed Anal. 2017 Jun 5;140:327-333. doi: 10.1016/j.jpba.2017.03.050. Epub 2017 Apr 1.
Acyl chlorides are important acylating agents in the synthesis of active pharmaceutical ingredients. Determining the residual acyl chlorides in drug substances is a challenge due to their high reactivity and the matrix interferences from drug substances and their related impurities. This paper describes a general derivatization HPLC method for the determination of aromatic and aliphatic acyl chlorides in lipophilic drug substances. Since most drug substances have weak absorptions in the visible range (above 380nm), the nitro-substituted anilines and nitro-substituted phenylhydrazines were selected as the derivatization reagents due to their weak basicity and red-shift of UV absorption spectra. The maximum wavelength and absorption intensity of nitro-substituted anilines decreased after derivatization with acyl chlorides, whereas the derivatization products of nitro-substituted phenylhydrazines showed the slight increases of maximum wavelength and absorbance intensity. Hence, 2-nitrophenylhydrazine was selected as the suitable derivatization reagent because the derivatives have the maximum UV wavelength absorbance at 395nm, which could largely minimize the matrix interferences. The optimization of the concentration of 2-nitrophenylhydrazine is important for the sensitivity and stability of derivatives. Other reaction conditions including reaction temperature, time and the influence of three competitive solvents (water, methanol and ethanol) on the reaction efficiency were also studied. After derivatization with 100μgmL 2-nitrophenylhydrazine at room temperature for 30min, the method was validated for high specificity and sensitivity with the detection limits in the range of 0.01-0.03μgmL. The proposed method was applied as a generic method to determine the residual acyl chlorides in lipophilic drug substances.
酰氯是活性药物成分合成中重要的酰化剂。由于其高反应活性以及药物物质及其相关杂质的基质干扰，测定药物物质中的残留酰氯是一项挑战。本文描述了一种用于测定亲脂性药物物质中芳香族和脂肪族酰氯的通用衍生化高效液相色谱法。由于大多数药物物质在可见光范围（380nm以上）吸收较弱，因此选择硝基取代苯胺和硝基取代苯肼作为衍生化试剂，因其碱性较弱且紫外吸收光谱发生红移。硝基取代苯胺与酰氯衍生化后，其最大波长和吸收强度降低，而硝基取代苯肼的衍生化产物最大波长和吸光度强度略有增加。因此，选择2-硝基苯肼作为合适的衍生化试剂，因为其衍生物在395nm处具有最大紫外波长吸光度，这可以在很大程度上减少基质干扰。优化2-硝基苯肼的浓度对于衍生物的灵敏度和稳定性很重要。还研究了其他反应条件，包括反应温度、时间以及三种竞争溶剂（水、甲醇和乙醇）对反应效率的影响。在室温下用100μg/mL 2-硝基苯肼衍生化30分钟后，该方法经验证具有高特异性和灵敏度，检测限在0.01 - 0.03μg/mL范围内。所提出的方法作为一种通用方法用于测定亲脂性药物物质中的残留酰氯。
