UPLC-MS/MS 方法测定醒脑静注射液中5 种成分的含量与方法研究

Contents of 5 components in Xingnaojing Injection were determined by UPLC-MS/MS method

关键词:  醒脑静注射液  UPLC-MS/MS  含量测定

Key words:  Xingnaojing Injection  UPLC-MS/MS  content determination

摘要:

[目的] 采用超高液相色谱-串联质谱（UPLC-MS/MS）技术建立可同时测定醒脑静注射液中莪术二酮、吉马酮、左旋樟脑、姜黄素和麝香酮5种指标性成分含量的新方法。[方法] 研究项下使用的色谱柱为ACQUITY HSS誖T3C18柱（2.1 mm×100 mm，1.8 μm），流动相A为0.1%甲酸水溶液+2 mmol乙酸铵，流动相B为甲醇溶液+0.1%甲酸。梯度洗脱方法： 0～0.5 min，95% A；0.5～1.0 min，95%→5% A；1.0～2.7 min，5% A；2.7～3.4 min，5%→95% A；3.4～4.0 min，95% A）。流速： 0.4 mL/min；柱温： 40℃；进样量： 5 μL；采用电喷雾离子源进行正离子监测，多反应监测模式（MRM）用于定量分析。[结果] 在4 min内，醒脑静注射液的5个标志性成分，包括莪术二酮、吉马酮、左旋樟脑、姜黄素和麝香酮被完全分离，峰面积与浓度呈现出良好的线性关系，相关系数（r）均大于0.998 0。对该检测方法的精密度、重复性和稳定性进行评价，均得到了满意结果。平均加样回收率为98.2%～104.2%。6批样品中莪术二酮、吉马酮、左旋樟脑、姜黄素和麝香酮含量测定结果分别为19.4～20.4、1.6～1.8、29.0～31.0、1.3～1.4、11.6～12.4 μg/mL。[结论] 该检测方法具有高灵敏度、准确可靠的优势，可实现对醒脑静注射液中多个组分快速检测，为该制剂的质量控制研究提供了参考。

Abstract:

[Objective] A new method for simultaneous determination of 5 index components containing curcuma dione，gemarone，L-camphor，curcumin and muscosterone in Xingnaojing Injection was established by ultra high liquid chromatography-tandem masss pectrometry（UPLC-MS/MS）. [Methods] The chromatographic column used in this research was ACQUITY HSS^? T3 C₁₈ column（2.1 mm×100 mm，1.8 μm）. The mobile phase consisted of 0.1% formic acid water and 2 mmol ammonium acetate（A），methanol and 0.1% formicacid（B） with gradient elution（0 to 0.5 min，95% A；0.5 to 1.0 min，95% to 5% A；1.0 to 2.7 min，5% A；2.7 to 3.4 min，5%→95% A；3.4 to 4.0 min，95% A）. The injection volume was 5 μL and the flow rate was 0.4 mL/min. The column temperature was 40 ℃. Electrospray ion source was adopted for positive ion monitoring，and multi-reaction monitoring mode（MRM） was used for quantitative analysis.[Results] Within 4 min，the five signature components of Xingnaojing Injection，including curcuma dione，jimarone，L-camphor，curcumin and muscatone，were completely separated. The peak area showed a good linear relationship with the concentration，and the correlation coefficient（r） was greater than 0.998 0. The accuracy，repeatability and stability of the method were evaluated，and satisfactory results were obtained. The average recoveries were 98.2% to 104.2%. The contents of curcuma dione，jimarone，L-camphor，curcumin and muscatone in 6 batches of samples were 19.4 to 20.4，1.6 to 1.8，29.0 to 31.0，1.3 to 1.4，11.6 to 12.4 μg/mL，respectively. [Conclusion] The method has the advantages of high sensitivity，accuracy and reliability，and can realize the rapid detection of multiple components in Xingnaojing Injection，which provides a reference for the quality control research of this preparation.