多中心切割2D-HPLC-QTOF法对表柔比星脂质体制剂中未知杂质的分离与结构推断  

Separation and inference of unknown impurities in epirubicin liposome by multiple heart-cutting 2D-HPLC-QTOF

作　　者：计小凡 欧阳辉[1] 冯育林[1] 饶毅 罗晓健[1] Ji Xiao-fan;Ouyang Hui;Feng Yu-lin;Rao Yi;Luo Xiao-jian(The National Pharmaceutical Engineering Center for Solid Preparation in Chinese Herbal Medicine,Jiangxi University of Chinese Medicine,Nanchang 330006)

机构地区：[1]江西中医药大学中药固体制剂制造技术国家工程研究中心,南昌330006

出　　处：《中国抗生素杂志》2023年第1期79-86,共8页Chinese Journal of Antibiotics

基　　金：江西省创新人才团队(No.20165BCB19009);南昌市高层次科技创新人才“双百计划”;江西中医药大学校级科技创新团队发展计划;国家科技重大专项(No.2017ZX09210004004)。

摘　　要：目的建立多中心切割(MHC)二维高效液相色谱-四级杆飞行时间质谱(2D-HPLC-QTOF)法分离和推断表柔比星脂质体制剂中未知杂质的方法。方法一维液相色谱采用Symmetry-C18(250 mm×4.6 mm,5μm)为色谱柱,以乙腈和甲醇(6:1)为流动相A,以加入十二烷基硫酸钠的磷酸缓冲液为流动相B,流动相A与流动相B体积比为57:43进行等度洗脱;二维高效液相色谱采用Agilent Eclipse-C18(50 mm×2.1 mm,1.8μm)为色谱柱,以0.1%的甲酸水和乙腈为流动相,采用梯度洗脱。将一维液相色谱条件下分离的杂质峰进行多中心切割储存在定量环中,并转移至二维进行分离,以四极杆飞行时间质谱(QTOF)进行结构推断。结果该方法可以使表柔比星与杂质峰较好地分离,并且可以得到高质量杂质峰进行结构推断。在表柔比星脂质体制剂中对6个未知杂质进行有效的推断。结论建立的多中心切割二维高效液相色谱-四极杆飞行时间质谱法可高效准确的分离并推断表柔比星脂质体制剂中的未知杂质,特别是微量和难以分离的杂质,对表柔比星制剂的质量控制提供了新方法。Objective To establish a method called multiple heart-cutting 2D-HPLC-QTOF to separate and identify the unknown impurities in epirubicin liposome.Methods One-dimension(1D)chromatographic condition was Symmetry-C18column using acetonitrile and methanol(6:1)as mobile phase A and phosphate buffer solution with sodium lauryl sulfate as mobile phase B.The volume ratio of mobile phase A to mobile phase B was 57:43.Two-dimension(2D)chromatographic condition was Agilent Eclipse-C18column using 0.1%formic acid solution as mobile phase A and acetonitrile as mobile phase B(gradient elution).Multiple impurity peaks separated by 1D liquid chromatography were cut by multi hearting-cutting and stored in quantitative loops,and then transferred to 2D for separation.The structures were identified by quadrupole time-of-flight mass spectrometry(QTOF).Results Using this method,good separation of epirubicin from impurity peaks was achieved,and high-quality impurity peaks were obtained for structural identification.Six unknown impurities were effectively identified in the epirubicin liposome.Conclusion The established method can efficiently and accurately separate and identify unknown impurities in the epirubicin liposome,especially the trace and hard-to-separate impurities,and provides a new method for quality control of the Epirubicin preparation.

关 键 词：2D-HPLC-QTOF 多中心切割 表柔比星脂质体 未知杂质 

分 类 号：R978.1[医药卫生—药品]