阿昔洛韦片中杂质的色谱-质谱结构分析

Structural Analysis of Impurities in Aciclovir Tablets by HPLC-MS/MS

关键词:  阿昔洛韦  四极杆-静电场轨道阱高分辨质谱  杂质  降解产物  结构鉴定

Key words:  aciclovir  Q-Exactive Orbitrap-MS  impurities  degradation products  structural identification

摘要:

目的 建立高效液相色谱-四级杆-静电场轨道阱高分辨质谱(HPLC-Q-Exactive Orbitrap-MS)联用法对阿昔洛韦片及其强制降解试验样品中的杂质进行结构分析。方法 采用Waters Xbridge BEH Shield RP₁₈(4.6 mm×250 mm，5 μm)色谱柱，以10 mmol·L^–1甲酸铵溶液(含0.15%甲酸)为流动相A，以10 mmol·L^–1甲酸铵溶液(含0.15%甲酸)-乙腈(50∶50)为流动相B，进行梯度洗脱，检测波长为254 nm，对阿昔洛韦片及其强制降解试验样品中的杂质进行分离，再采用Q-Exactive Orbitrap-MS测定各杂质一级精确分子量及二级碎片离子，并进行结构鉴定。结果 阿昔洛韦及其杂质均分离良好，检测并鉴定出阿昔洛韦片及其强制降解试验样品中8个含量>0.1%的主要杂质，其中4个为欧洲药典10.0规定的已知杂质，而其他杂质均未见报道。结论 建立的液质联用技术能有效鉴定阿昔洛韦片的杂质，能够为其生产工艺的优化和质量控制提供参考。

Abstract:

OBJECTIVE To analysis the structures of the impurities in aciclovir tablets and the compulsory degradation samples by high performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry technology (HPLC-Q-Exactive Orbitrap-MS). METHODS The HPLC separation was carried out on a Waters Xbridge BEH Shield RP₁₈ (4.6 mm×250 mm, 5 μm) by the gradient elution with a mobile phase consisting of 10 mmol·L^–1 ammonium formate solution (0.15% formic acid)(A) and 10 mmol·L^–1 ammonium formate solution(0.15% formic acid)-acetonitrile(50∶50)(B), and the detection wavelength was 254 nm. The Q-Exactive Orbitrap-MS was used to determine the precise first-order molecular weight and second-order fragment ions of these impurities, and the structures were identified. RESULTS Aciclovir and its impurities were well separated, and 8 major impurities with content>0.1% were detected and identified in aciclovir tablets and the compulsory degradation samples. Among them, 4 were the impurities listed in the European Pharmacopeia 10.0, and the others were unknown impurities identified for the first time. CONCLUSION The LC-MS/MS method can effectively identify the impurities in aciclovir tablets, which is significant for the production process optimization and quality control.