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

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,5μm)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QExactive 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。

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 secondorder 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.