赵钰馨 孙秉喆 倪卫星 梁然植 狄斌 苏梦翔

中圖分类号 R917 文献标志码 A 文章编号 1001-0408（2021）18-2189-05

DOI 10.6039/j.issn.1001-0408.2021.18.03

摘 要 目的：建立阿嗪米特原料药中潜在基因毒性杂质马来酰肼含量测定的方法。方法：采用高效液相色谱-荧光检测器法。以Thermo Syncronis C18为色谱柱，以0.2 mol/L乙酸溶液-甲醇为流动相进行梯度洗脱;柱温为30 ℃;激发波长为315 nm，发射波长为389 nm;流速为1 mL/min;进样量为20 μL。结果：空白溶剂和阿嗪米特均不干扰马来酰肼的测定;马来酰肼检测质量浓度的线性范围为19.5～300 ng/mL（r=0.999 9）;检测限为4.5 ng/mL，定量限为19.5 ng/mL;加样回收率为98.79%～103.76%（RSD均小于3.00%，n=9）;精密度、稳定性（24 h）试验的RSD均不大于5.63%，且耐用性试验的RSD均小于2.00%（n=6）。3批阿嗪米特原料药中均未检出马来酰肼。结论：该方法专属性好、灵敏度高、定量准确，可用于阿嗪米特或其他基质中马来酰肼的痕量测定。

关键词 马来酰肼;阿嗪米特原料药;高效液相色谱-荧光检测器法;基因毒性杂质

Content Determination of Potential Genotoxic Impurity Maleic Hydrazide in Azintamide

ZHAO Yuxin1，SUN Bingzhe1，2，NI Weixing2，LIANG Ranzhi2，DI Bin1，3，4，SU Mengxiang1，3，4[1. School of Pharmacy， China Pharmaceutical University， Nanjing 210009， China; 2. Yangzhou Yiyang Pharmaceutical Co.， Ltd.， Jiangsu Yangzhou 225600， China; 3. Key Laboratory of Drug Quality Control and Pharmacovigilance （China Pharmaceutical University）， Ministry of Education， Nanjing 210009， China; 4. Key Laboratory for Research and Evaluation of Pharmaceutical Preparations and Excipients， National Medical Products Administration， Nanjing 210009， China]

ABSTRACT   OBJECTIVE： To establish a method for the content determination of potential genotoxic impurity maleic hydrazide in azintamide raw material. METHODS： HPLC-FLD method was adopted. The determination was performed on Thermo Syncronis C18 column with mobile phase consisted of 0.2 mol/L acetic acid-methanol （gradient elution）. The column temperature was set at 30 ℃， the excitation wavelength was 315 nm and emission wavelength was 389 nm. The flow rate was 1 mL/min， and the sample size was 20 μL. RESULTS： The blank solvent and azintamide did not interfere with the determination of maleic hydrazide. The linear range of maleic hydrazide was 19.5-300 ng/mL （r=0.999 9）. The limit of detection was 4.5 ng/mL and the limit of quantification was 19.5 ng/mL. The recovery ranged from 98.79% to 103.76% （RSDs were lower than 3.00%，n=9）. RSDs of precision and stability（24 h）tests were no more than 5.63%，and those of durability tests were less than 2.00% （n=6）. Maleic hydrazide was not detected in 3 batches of azinamide raw material. CONCLUSIONS： The method is specific， sensitive and accurate. It can be used for the trace determination of maleic hydrazide in azintamide or other matrix.

KEYWORDS   Maleic hydrazide; Azintamide; HPLC-FLD; Genotoxic impurity

马来酰肼（maleic hydrazide）的化学名为顺丁烯二酰肼，是一种合成哒嗪类化合物的常用原料。哒嗪类化合物作为重要的中间体，被广泛应用于农药及头孢唑兰、普纳替尼等药品的合成中[1-4]。尽管目前尚无证据证明马来酰肼具有基因毒性和致癌性，但其结构中存在警示结构——肼，故该成分为潜在的基因毒性杂质[5]。潜在基因毒性杂质的毒性虽然未经实验证实或模型验证，但人用药品注册技术要求国际协调会、美国FDA及欧洲药品管理局等均对其在药品中的限度提出了具体要求[6-8]。

阿嗪米特（azintamide）的化学名为2-（6-氯哒嗪-3-基）硫烷基-N，N-二乙基乙酰胺，于1959年首次合成[9]，是一种促进胆汁分泌的药物，可用于治疗因胆汁分泌不足而引起的消化系统方面的疾病[10-11]。……