联用技术鉴定与测定硫酸阿托品有关物质
Identification and determination of related substances in atropine sulfate with hyphenated technique
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摘要: 建立高效液相离子对色谱法测定硫酸阿托品的有关物质，并利用LC-MS/MS进行结构鉴定。采用十八烷基硅烷键合硅胶为填充剂，以磷酸二氢钾缓冲液(含庚烷磺酸钠，用磷酸调pH至5.0)-乙腈为流动相进行线性梯度洗脱，对硫酸阿托品及其碱、高温、氧化破坏的有关物质进行分离；并在挥发性流动相条件下通过高分辨LC-TOF/MS测定各有关物质的准确质量和LC-MS/MS二级质谱测定鉴定结构。建立的离子对色谱条件下各有关物质与主成分分离良好，检测到12个主要有关物质，其中6个为欧洲药典已知杂质，6个为国内外文献未见报道的未知杂质，经初步鉴定它们为硫酸阿托品的合成副产物和降解产物。新建立的离子对色谱法适用于硫酸阿托品中有关物质的检查，为硫酸阿托品质量控制和工艺优化提供了参考。Abstract: A specific ion-pairing HPLC method was developed for the determination of related substances in atropine sulfate. Their structures were identified by LC-MS/MS. HPLC separation was carried out on a Thermo BDS Hypersil C18 column with linear gradient elution by a mobile phase consisting of acetonitrile and phosphate buffer solution(containing sodium heptane-sulphonate, adjusting pH to 5. 0 with phosphoric acid). The flow rate was 0. 8 mL/min and UV detection wavelength was 225 nm. Atropine sulfate and its related substances can be completely separated under the established HPLC condition. And they were characterized by hyphenated chromatographic techniques with accurate mass determination using high resolution LC-TOF/MS methods as well as product MS spectra scanning through a volatile mobile phase. Twelve related substances were detected and identified as by-products from synthesis and degradations of atropine sulfate. Six of them were related substances from EP and the other six were unknown related substances. This newly established HPLC method is effective in the separation and identification of the related substances in atropine sulfate and the results are useful for its quality control and process optimization.
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Keywords:
- atropine sulfate /
- related substances /
- HPLC /
- LC-MS/MS
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