二维超高效液相色谱-Qtof质谱联用技术在国产与原研注射用盐酸万古霉素杂质对比中的应用

Application of two-dimensional UPLC-Qtof MS technoligy in the study of impurity profile of domestic product vancomycin hydrochloride for injection and reference listed drug

目的建立二维超高效液相色谱-Qtof质谱联用研究注射用盐酸万古霉素杂质谱的方法,对比仿制制剂与参比制剂杂质谱一致性。方法通过二维液相色谱在线脱盐技术,在不改变一维液相色谱条件的基础上,将目标化合物通过阀切换转换至二维液相色谱柱,在二维液相流动相的带动下进质谱,进行结构鉴定。一维色谱柱为ACQUITY UPLC CSHTM C_(18)(150 mm×2.1 mm,1.7μm),流动相A为乙腈-甲醇-溶液A(3∶4∶93,V/V/V),流动相B为乙腈-甲醇-溶液A(10∶40∶50,V/V/V),溶液A为称取7.0 g Tris至约950 ml水中,经20%乙酸溶液(V/V)调节pH至约8.22,用水定容至1000 ml制得,检测波长为280 nm;二维液相色谱柱为ACQUITY UPLC BEH C_(18)(50 mm×2.1 mm,1.7μm),流动相A为0.1%甲酸水溶液,流动相B为乙腈;采用ESI离子化模式进行质谱扫描,离子源温度120℃,雾化气温度500℃,雾化气流速900 L/h,采集模式为MSE及MSMS。结果对样品中存在的杂质进行分析,确认9个已知杂质与参比制剂结构一致,解析出可能的未知杂质结构8个,其结构与参比制剂一致。结论本法建立了二维超高效液相色谱-Qtof质谱联用技术测定、分析注射用盐酸万古霉素杂质的方法,可快速实现杂质定性,结合软件解析结构更便捷;国产制剂与参比制剂产品相比,杂质谱基本一致,表明该仿制制剂与参比制剂杂质水平相当。

Objective To establish a two-dimensional UPLC-QTof MS method for the study of impurity profile of Vancomycin Hydrochloride for Injection.To compare the consistency of impurity profile between generic preparation and reference listed drug(RLD).Methods By two-dimensional liquid chromatography online desalination technology,there was no need to change the first-dimensional liquid chromatography conditions,and converse the target compound to two-dimensional liquid chromatography column by valve switching,which led it into the mass spectrometer under the seconddimensional mobile phase elution,and the compound structure was identified.First-dimensional column was Inertsil ACQUITY UPLC CSHTM C_(18)(150 mm×2.1 mm,1.7μm),the mobile phase A was acetonitrile-methyl alcohol-solution A(3∶4∶93,v/v/v),the mobile phase B was acetonitrile-methyl alcohol-solution A(10∶40∶50,v/v/v),the solution A:dissolve 7.0 g tris(hydroxymethyl)aminomethane R in approximately 950 ml of waters for chromatography R,detection wavelength:280 nm;seconddimensional column was ACQUITY UPLC BEH C_(18)(50 mm×2.1 mm,1.7μm),mobile phase A was 0.1% formic acid solution,mobile phase B was acetonitrile,gradient elution.Electrospray ionization(ESI)source was used as negative ion detection model,the temperature of ESI source was 120℃,nebulizing temperature was 500℃,nebulizing flow rate was 900 L/h,and the acquisition mode was MSE and MSMS.Results The existing impurities in the sample were determined,nine known impurities of determined were consistent with the structure of the RLD,and eight possible impurities structure were obtained.Conclusion This article was focused on the method establishment to determine Vancomycin Hydrochloride for Injection impurities by two-dimensional UPLC-QTof MS technology.The qualitative of impurity was quickly obtained by using 2-dimensional HPLC-MS,and it is convenient to identify the structure of impurities combining the software.The domestic products’impurity profile was the same as that of the RLD,indicating that the quality of the generic drug is equivalent to that of the RLD.